Opa1 antisense oligomers for treatment of conditions and diseases

Abstract

Alternative splicing events in genes can lead to nonproductive mRNA transcripts which in turn can lead to aberrant protein expression, and therapeutic agents which can target the alternative splicing events in genes can modulate the expression level of functional proteins in patients and / or inhibit aberrant protein expression. Such therapeutic agents can be used to treat a condition or disease caused by protein deficiency and / or mitochondrial function deficit.

Description

OPA1 ANTISENSE OLIGOMERS FOR TREATMENT OF CONDITIONS AND DISEASES CROSS-REFERENCE

[0001] This application claims benefit of United States Provisional Patent Application No. 63 / 505,333 filed May 31, 2023, which is incorporated herein by reference in its entirety.BACKGROUND

[0002] Alternative splicing events in genes can lead to nonproductive mRNA transcripts which in turn can lead to aberrant or reduced protein expression, and therapeutic agents which can target the alternative splicing events in genes can modulate the expression level of functional proteins in patients and / or inhibit aberrant protein expression. Such therapeutic agents can be used to treat a condition or disease caused by the protein deficiency.

[0003] Autosomal dominant optic atrophy (ADOA) is one of the most commonly diagnosed optic neuropathies. This optic nerve disease is associated with structural and functional mitochondrial deficits that lead to degeneration of the retinal ganglion cells and progressive, irreversible loss of vision. A majority of ADOA patients carry mutations in OPA1 and most mutations lead to haploinsufficiency (Lenaers G. et al. Orphanet J Rare Dis 2012). OPA1 encodes a mitochondrial GTPase with a critical role in mitochondrial fusion, ATP synthesis and apoptosis. Currently, there is no approved disease-modifying treatment for ADOA patients and there is a need for such treatments.SUMMARY

[0004] Described herein, in some aspects, is a method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

[0005] Described herein, in some aspects, is a method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299,wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by having:(i) a heterozygous OPA1 gene variant;(ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments;(iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity;(iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or(v) any combination of (i)-(iv).

[0006] Described herein, in some aspects, is a method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by:(1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene;(2) not having only benign or likely benign variants in the OPA1 gene;(3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA- plus);(4) not having been diagnosed with Behr syndrome;(5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases;(6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy;(7) not having or having a history of any ocular condition in either eye;(8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering;(9) not having a history of retinal photocoagulation;(10) not having a history or presence of retinal vein occlusion;(11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering;(12) not having dry age-related macular degeneration in either eye;(13) not having high myopia (>6 diopters);(14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated);(15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy;(16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery; or(17) any combination of (1)-(16).

[0007] Described herein, in some aspects, is a method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0008] In some aspects, the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

[0009] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mg to about 15 mg, about 0.01 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg,about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

[0010] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

[0011] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

[0012] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer.

[0013] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL to about 250 pL, about 10 pL to about 250 pL, about 20 pL to about 250 pL, about 30 pL to about 250 pL, about 40 pL to about 250 pL,about 50 pL to about 250 pL, about 60 pL to about 250 pL, about 70 pL to about 250 pL, about 80 pL to about 250 pL, about 100 pL to about 250 pL, about 120 pL to about 250 pL, about 150 pL to about 250 pL, about 160 pL to about 250 pL, about 180 pL to about 500 pL, about 200 pL to about 250 pL, about 220 pL to about 250 pL, about 5 pL to about 220 pL, about 10 pL to about 220 pL, about 20 pL to about 220 pL, about 30 pL to about 220 pL, about 40 pL to about 220 pL, about 50 pL to about 220 pL, about 60 pL to about 220 pL, about 70 pL to about 220 pL, about 80 pL to about 220 pL, about 100 pL to about 220 pL, about 120 pL to about 220 pL, about 150 pL to about 220 pL, about 160 pL to about 220 pL, about 180 pL to about 220 pL, about 5 pL to about 200 pL, about 10 pL to about 200 pL, about 20 pL to about 200 pL, about 30 pL to about 200 pL, about 40 pL to about 200 pL, about 50 pL to about 200 pL, about 60 pL to about 200 pL, about 70 pL to about 200 pL, about 80 pL to about 200 pL, about 100 pL to about 200 pL, about 120 pL to about 200 pL, about 150 pL to about 200 pL, about 160 pL to about 200 pL, about 180 pL to about 200 pL, about 5 pL to about 180 pL, about 10 pL to about 180 pL, about 20 pL to about 180 pL, about 30 pL to about 180 pL, about 40 pL to about 180 pL, about 50 pL to about 180 pL, about 60 pL to about 180 pL, about 70 pL to about 180 pL, about 80 pL to about 180 pL, about 100 pL to about 180 pL, about 120 pL to about 180 pL, about 150 pL to about 180 pL, about 5 pL to about 150 pL, about 10 pL to about 150 pL, about 20 pL to about 150 pL, about 30 pL to about 150 pL, about 40 pL to about 150 pL, about 50 pL to about 150 pL, about 60 pL to about 150 pL, about 70 pL to about 150 pL, about 80 pL to about 150 pL, about 100 pL to about 150 pL, about 120 pL to about 150 pL, about 5 pL to about 150 pL, about 10 pL to about 120 pL, about 20 pL to about 120 pL, about 30 pL to about 120 pL, about 40 pL to about 120 pL, about 50 pL to about 120 pL, about 60 pL to about 120 pL, about 70 pL to about 120 pL, about 80 pL to about 120 pL, about 100 pL to about 120 pL, about 5 pL to about 100 pL, about 10 pL to about 100 pL, about 20 pL to about 100 pL, about 30 pL to about 100 pL, about 40 pL to about 100 pL, about 50 pL to about 100 pL, about 60 pL to about 100 pL, about 70 pL to about 100 pL, about 80 pL to about 100 pL, about 5 pL to about 80 pL, about 10 pL to about 80 pL, about 20 pL to about 80 pL, about 30 pL to about 80 pL, about 40 pL to about 80 pL, about 50 pL to about 80 pL, about 60 pL to about 80 pL, about 5 pL to about 60 pL, about 10 pL to about 60 pL, about 20 pL to about 60 pL, about 30 pL to about 60 pL, about 40 pL to about 60 pL, or about 50 pL to about 60 pL.

[0014] In some aspects, the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL, about 8 pL, about 10 pL, about 12 pL, about 15 pL, about 18 pL, about 20 pL, about 25 pL, about 28 pL, about 30 pL, about 35 pL, about 40 pL, about 45 pL, about 48 pL, about 50 pL, about 55 pL, about 60 pL, about 65 pL,about 70 pL, about 75 pL, about 80 pL, about 90 pL, about 100 pL, about 120 pL, about 150 pL, about 160 pL, about 180 pL, about 200 pL, about 220 pL, or about 250 pL.

[0015] In some aspects, the method comprises administering the pharmaceutical composition to both left eye and right eye of the subject.

[0016] In some aspects, the method comprises administering the pharmaceutical composition at the same dose to both the left eye and the right eye of the subject.

[0017] In some aspects, the method comprises administering the pharmaceutical composition at different doses to the left eye and the right eye of the subject.

[0018] In some aspects, the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to the sequence set forth in any one of SEQ ID NOs: 6-275 or 280-299.

[0019] In some aspects, the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to the sequence set forth in any one of SEQ ID NO: 36, 236, 242, 250, 280-283, 288, or 290-292.

[0020] In some aspects, the antisense oligomer modulates splicing of a non-sense mediated RNA decay -inducing exon (NMD exon) from a pre-mRNA in a cell of the subject, wherein the pre- mRNA encodes the OPA1 protein and comprises the NMD exon, thereby modulating a level of processed mRNA that is processed from the pre-mRNA, and modulating expression of the OPA1 protein in the cell.

[0021] In some aspects, the antisense oligomer:(a) binds to a targeted portion of the pre-mRNA;(b) modulates binding of a factor involved in splicing of the NMD exon; or(c) a combination of (a) and (b).

[0022] In some aspects, the targeted portion of the pre-mRNA is proximal to the NMD exon.

[0023] In some aspects, the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of 5’ end of the NMD exon.

[0024] In some aspects, the targeted portion of the pre-mRNA is at least about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides, about 40 nucleotides, about 30 nucleotides, about 20 nucleotides, about 10nucleotides, about 5 nucleotides, about 4 nucleotides, about 2 nucleotides, about 1 nucleotides upstream of 5’ end of the NMD exon.

[0025] In some aspects, the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of 3’ end of the NMD exon.

[0026] In some aspects, the targeted portion of the pre-mRNA is at least about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides, about 40 nucleotides, about 30 nucleotides, about 20 nucleotides, about 10 nucleotides, about 5 nucleotides, about 4 nucleotides, about 2 nucleotides, about 1 nucleotides downstream of 3’ end of the NMD exon.

[0027] In some aspects, the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193628509.

[0028] In some aspects, the targeted portion of the pre-mRNA is about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193628509.

[0029] In some aspects, the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193628616.

[0030] In some aspects, the targeted portion of the pre-mRNA is about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193628616.

[0031] In some aspects, the targeted portion of the pre-mRNA is located in an intronic region between two canonical exonic regions of the pre-mRNA, and wherein the intronic region contains the NMD exon.

[0032] In some aspects, the targeted portion of the pre-mRNA at least partially overlaps with the NMD exon.

[0033] In some aspects, the targeted portion of the pre-mRNA at least partially overlaps with an intron upstream or downstream of the NMD exon.

[0034] In some aspects, the targeted portion of the pre-mRNA comprises 5’ NMD exon-intron junction or 3’ NMD exon-intron junction.

[0035] In some aspects, the targeted portion of the pre-mRNA is within the NMD exon.

[0036] In some aspects, the targeted portion of the pre-mRNA comprises about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more consecutive nucleotides of the NMD exon.

[0037] In some aspects, the NMD exon comprises a sequence with at least 80%, at least 90%, or 100% sequence identity to SEQ ID NO: 279.

[0038] In some aspects, the NMD exon comprises a sequence of SEQ ID NO: 279.

[0039] In some aspects, the targeted portion of the pre-mRNA is within the non-sense mediated RNA decay-inducing exon GRCh38 / hg38: chr3 193628509 to 193628616.

[0040] In some aspects, the targeted portion of the pre-mRNA is upstream or downstream of the non-sense mediated RNA decay-inducing exon GRCh38 / hg38: chr3 193628509 to 193628616.

[0041] In some aspects, the targeted portion of the pre-mRNA comprises an exon-intron junction of exon GRCh38 / hg38: chr3 193628509 to 193628616.

[0042] In some aspects, the OPA1 protein expressed from the processed mRNA is a full-length OPA1 protein or a wild-type OPA1 protein.

[0043] In some aspects, the OPA1 protein expressed from the processed mRNA is a functional OPA1 protein.

[0044] In some aspects, the OPA1 protein expressed from the processed mRNA is at least partially functional as compared to a wild-type OPA1 protein.

[0045] In some aspects, the OPA1 protein expressed from the processed mRNA is at least partially functional as compared to a full-length wild-type OPA1 protein.

[0046] In some aspects, the antisense oligomer promotes exclusion of the NMD exon from the pre-mRNA.

[0047] In some aspects, the exclusion of the NMD exon from the pre-mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5- fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

[0048] In some aspects, the method results in an increase in the level of the processed mRNA in the cell.

[0049] In some aspects, the level of the processed mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

[0050] In some aspects, the method results in an increase in the expression of the OPA1 protein in the cell.

[0051] In some aspects, a level of the OPA1 protein expressed from the processed mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5- fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

[0052] In some aspects, the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

[0053] In some aspects, the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N- methyl-acetamide (2’-NMA), or a 2’-O-methoxyethyl moiety.

[0054] In some aspects, the antisense oligomer comprises at least one modified sugar moiety.

[0055] In some aspects, each sugar moiety is a modified sugar moiety.

[0056] In some aspects, the antisense oligomer comprises a 5 ’-methylcytosine (5’-MeC).

[0057] In some aspects, each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’-MeC).

[0058] In some aspects, the antisense oligomer comprises a 5 ’-methyluracil (5’-MeU).

[0059] In some aspects, the antisense oligomer comprises a phosphorothioate linkage.

[0060] In some aspects, each internucleoside linkage of the ASO is a phosphorothioate linkage.

[0061] In some aspects, the antisense oligomer comprises a locked nucleic acid (LNA).

[0062] In some aspects, the antisense oligomer is from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases,9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases in length.

[0063] In some aspects, the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0064] In some aspects, the antisense oligomer is a compound according to any of the following structures:

[0065] In some aspects, the pharmaceutical composition is a liquid composition.

[0066] In some aspects, the method comprises administering the pharmaceutical composition as a bolus injection.

[0067] In some aspects, the method comprises administering the pharmaceutical composition as a bolus injection over 1 to 60 minutes, 1 to 50 minutes, 1 to 40 minutes, 1 to 30 minutes, 1 to 20 minutes, 1 to 10 minutes, 1 to 5 minutes, or 1 to 3 minutes.

[0068] In some aspects, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for inj ection.

[0069] In some aspects, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for inj ection.

[0070] In some aspects, the antisense oligomer is solubilized or diluted in an isotonic solution.

[0071] In some aspects, the antisense oligomer is solubilized or diluted in a phosphate-buffered solution with at least pH 5.8.

[0072] In some aspects, the antisense oligomer is solubilized or diluted in a phosphate-buffered (pH 6.6 - 7.6) solution.

[0073] In some aspects, the pharmaceutical formulation does not comprise a preservative.

[0074] In some aspects, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL.

[0075] In some aspects, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL, about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

[0076] In some aspects, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0077] In some aspects, the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer.

[0078] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL toabout 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0079] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0080] In some aspects, the concentrate is a phosphate-buffered solution.

[0081] In some aspects, the subject is a human subject.

[0082] In some aspects, the disease or condition is associated with a deficient amount or activity of the OPA1 protein.

[0083] In some aspects, the disease or condition comprises an eye disease or condition.

[0084] In some aspects, the disease or condition comprises a cardiovascular disease or condition.

[0085] In some aspects, the disease or condition comprises a neurological disease or condition.

[0086] In some aspects, the disease or condition comprises ADOA-plus syndrome; a mitochondrial disorder; glaucoma; normal tension glaucoma; Charcot-Mari e-Tooth disease; mitochondria dysfunction; diabetic retinopathy; age-related macular degeneration; retinal ganglion cell death; mitochondrial fission-mediated mitochondrial dysfunction; progressive external ophthalmoplegia; deafness; ataxia; motor neuropathy; sensory neuropathy; myopathy; Behr syndrome; brain dysfunction; encephalopathy; peripheral neuropathy; fatal infantile mitochondrial encephalomyopathy; hypertrophic cardiomyopathy; spastic ataxic syndrome; sensory motor peripheral neuropathy; hypotonia; gastrointestinal dysmotility and dysphagia; optic atrophy; optic atrophy plus syndrome; Mitochondrial DNA depletion syndrome 14; late-onset cardiomyopathy; diabetic cardiomyopathy; Alzheimer’s Disease; focal segmental glomerulosclerosis; kidney disease; Huntington’s Disease; cognitive function decline in healthy aging; Prion diseases; late onset dementia and parkinsonism; mitochondrial myopathy; Leigh syndrome; Friedreich’s ataxia; Parkinson’s disease; MELAS (Mitochondrial encephalomyopathy,lactic acidosis, and stroke-like episodes); pyruvate dehydrogenase complex deficiency; chronic kidney disease; Leber’s hereditary optic neuropathy; obesity; age-related systemic neurodegeneration; skeletal muscle atrophy; heart and brain ischemic damage; or massive liver apoptosis.

[0087] In some aspects, the disease or condition comprises Optic atrophy type 1.

[0088] In some aspects, the disease or condition comprises autosomal dominant optic atrophy (ADOA).

[0089] In some aspects, the subject is characterized by having:(i) a heterozygous OPA1 gene variant;(ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments;(iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity;(iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or(v) any combination of (i)-(iv).

[0090] In some aspects, the subject is additionally characterized by:(1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene;(2) not having only benign or likely benign variants in the OPA1 gene;(3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA- plus);(4) not having been diagnosed with Behr syndrome;(5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases;(6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy;(7) not having or having a history of any ocular condition in either eye;(8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering;(9) not having a history of retinal photocoagulation;(10) not having a history or presence of retinal vein occlusion;(11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering;(12) not having dry age-related macular degeneration in either eye;(13) not having high myopia (>6 diopters);(14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated);(15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy;(16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery;(17) any combination of (1)-(16).

[0091] In some aspects, administering comprises administering multiple doses of the pharmaceutical composition to the human subject.

[0092] In some aspects, administering comprises administering a first dose of the pharmaceutical composition to the human subject and a subsequent dose of the pharmaceutical composition to the human subj ect.

[0093] In some aspects, the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is not tolerated.

[0094] In some aspects, the subsequent dose is the same as the previous dose following an indication that administration of the previous dose is tolerated.

[0095] In some aspects, the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is tolerated.

[0096] In some aspects, the subsequent dose is the same as the previous dose following an indication that administration of the previous dose is effective.

[0097] In some aspects, the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is effective.

[0098] In some aspects, the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is not effective.

[0099] In some aspects, the pharmaceutical composition is administered via intracerebroventricular injection, intraperitoneal injection, intramuscular injection, intrathecal injection, subcutaneous injection, oral administration, synovial injection, intravitreal administration, subretinal injection, topical application, implantation, or intravenous injection.

[0100] In some aspects, the pharmaceutical composition is administered via intravitreal injection.

[0101] In some aspects, the method further comprises administering an additional therapeutic agent.

[0102] In some aspects, the additional therapeutic agent comprises a small molecule.

[0103] In some aspects, the additional therapeutic agent comprises an antisense oligomer.

[0104] In some aspects, the additional therapeutic agent comprises an ophthalmologic drug.

[0105] Described herein, in some aspects, is an antisense oligomer that is a compound according to any one of the following structures:or a pharmaceutically acceptable salt thereof.

[0106] Described herein, in some aspects, is an antisense oligomer having any of the following structures:

[0107] Described herein, in some aspects, is a pharmaceutical formulation comprising:(a) an antisense oligomer, wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution at a concentration of about 1 mg / mL to about 200 mg / mL.

[0108] Described herein, in some aspects, is a pharmaceutical formulation comprising:(a) an antisense oligomer; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, andwherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0109] Described herein, in some aspects, is a pharmaceutical formulation comprising:(a) an antisense oligomer; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, and wherein the antisense oligomer has any one of the following chemical structures:

[0110] In some aspects, the antisense oligomer is present in the solution at a concentration of about 1 mg / mL to about 200 mg / mL.

[0111] In some aspects, the antisense oligomer is present in the solution at a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

[0112] In some aspects, the antisense oligomer is present in the solution at a concentration of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0113] In some aspects, the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer.

[0114] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL,about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0115] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0116] In some aspects, the concentrate is phosphate buffered.

[0117] In some aspects, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection.

[0118] In some aspects, the antisense oligomer is solubilized or diluted in a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for inj ection.

[0119] In some aspects, the antisense oligomer is solubilized or diluted in an isotonic solution.

[0120] In some aspects, the antisense oligomer is solubilized or diluted in a phosphate- buffered solution with at least pH 5.8.

[0121] In some aspects, the antisense oligomer is solubilized or diluted in a phosphate- buffered (pH 6.6 - 7.6) solution.

[0122] In some aspects, the pharmaceutical formulation does not comprise a preservative.

[0123] In some aspects, the pharmaceutical formulation is suitable for an intravitreal injection.

[0124] In some aspects, the pharmaceutical formulation is packaged in a single-use vial.

[0125] Described herein, in some aspects, is a kit comprising:(i) a concentrate comprising an antisense oligomer (ASO), wherein the ASO comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280- 299; and(ii) a diluent, wherein the concentrate is miscible with the diluent; and(iii) instructions for diluting the concentrate with the diluent.

[0126] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL toabout 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0127] In some aspects, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0128] In some aspects, the concentrate is phosphate buffered.

[0129] In some aspects, the diluent is a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection.

[0130] In some aspects, the diluent is a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection.

[0131] In some aspects, the diluent comprises an isotonic solution.

[0132] In some aspects, the diluent comprises a phosphate-buffered solution with at least pH 5.8.

[0133] In some aspects, the diluent comprises a phosphate-buffered (pH 6.6 - 7.6) solution.

[0134] In some aspects, the concentrate or the diluent does not comprise a preservative.

[0135] In some aspects, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 2 mg / mL to 200 mg / mL in the diluent.

[0136] In some aspects, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL in the diluent.

[0137] In some aspects, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the antisense oligomer to a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL in the diluent.

[0138] In some aspects, the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299.

[0139] In some aspects, the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292.

[0140] In some aspects, the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

[0141] In some aspects, the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N-methyl-acetamide (2’-NMA), or a 2’-O-methoxyethyl moiety.

[0142] In some aspects, the antisense oligomer comprises at least one modified sugar moiety.

[0143] In some aspects, each sugar moiety is a modified sugar moiety.

[0144] In some aspects, the antisense oligomer comprises a 5 ’-methylcytosine (5’-MeC).

[0145] In some aspects, each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’- MeC).

[0146] In some aspects, the antisense oligomer comprises a 5 ’-methyluracil (5’-MeU).

[0147] In some aspects, the antisense oligomer comprises a phosphorothioate linkage.

[0148] In some aspects, each intemucleoside linkage of the ASO is a phosphorothioate linkage.

[0149] In some aspects, the antisense oligomer comprises a locked nucleic acid (LNA).

[0150] In some aspects, the antisense oligomer is from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases in length.

[0151] In some aspects, the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0152] In some aspects, the antisense oligomer has any one of the following chemical structures:

[0153] Described herein, in some aspects, are uses of an antisense oligomer for the manufacture of a medicament for treating a disease or condition characterized by a reduced expression or function of OPA1 protein in a human subject in need thereof, or reducing likelihood of the subject developing the disease or condition, wherein the medicament is administered to one eye of the subject at a dose of about 0.005 mg to about 20 mg, and wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299.

[0154] In some aspects, the medicament is administered to the one eye of the subject at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mg to about 15 mg, about 0.01 mg to about 10mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

[0155] In some aspects, the medicament is administered to the one eye of the subject at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

[0156] In some aspects, the medicament is administered to the one eye of the subject at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about 1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

[0157] In some aspects, the medicament is administered to the one eye of the subject at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer.INCORPORATION BY REFERENCE

[0158] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0159] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0160] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:

[0161] FIG. 1A depicts a schematic representation of a target mRNA that contains a nonsense-mediated mRNA decay-inducing exon (NMD exon mRNA) and therapeutic agent- mediated exclusion of the nonsense-mediated mRNA decay-inducing exon to increase expression of the full-length target protein or functional RNA. In particular, FIG. 1A shows an example schematic of the inclusion of an alternative exon(designated as Exon X) identified in the OP Al gene which leads to the introduction of a premature termination codon (PTC) resulting in a nonproductive mRNA transcript degraded by nonsense-mediated decay (NMD). The schematic also shows the location of the two primers used to detect both the productive and nonproductive mRNA transcripts by RT-PCR.

[0162] FIG. IB illustrates expression of OPA1 transcripts containing the NMD exon in HEK293 cells treated with increasing amounts of cycloheximide. Treatment of HEK293 cells with cycloheximide for 3 hours increases the abundance of nonproductive mRNA transcripts. Nonproductive transcript abundance was calculated as the intensity of the (nonproductive transcript / [the nonproductive + productive transcript]) and expressed as a percentage. A representative gel image is shown, and the percentage abundance of the nonproductive transcript indicated below is an average of three biological replicates.

[0163] FIG. 1C shows RT-PCR data from nonproductive transcripts in RNA isolated from retinal tissue punches from macular and peripheral regions of both eyes of a human donor. The relative abundance of the nonproductive transcript in each sample is included at the bottom of each lane. OD: oculus dexter (right eye), OS: oculus sinister (left eye).

[0164] FIG. 2A shows RT-PCR data for nonproductive OPA1 mRNAs in HEK293 cells after treatment with ASO-1 and cycloheximide in vitro.

[0165] FIG. 2B illustrates quantification of productive OPA1 mRNAs in HEK293 cells after treatment with ASO-1 in the absence of cycloheximide.

[0166] FIG. 2C illustrates protein expression of OPA1 in HEK293 cells after treatment with ASO-1 in the absence of cycloheximide.

[0167] FIG. 2D is an EC50 evaluation of ASO-1 in HEK293 cells and illustrates a representative dose-response curve fit based on levels of nonproductive transcript measured when various doses of ASO-1 were applied to HEK293 cells.

[0168] FIG. 2E are Western Blot gel images showing OPA1 protein expression after the transfection of HEK293 cells with vehicle (first three lanes), a non-targeting ASO (NT ASO) at various concentrations (1 nM, 5 nM, and 20 nM), and ASO-1 at various concentrations (1 nM, 5 nM, and 20 nM). Each condition comprised six biological replicates. Cells were lysed with RIPA buffer after transfection and Western Blots were probed with antibodies targeting OPA1 and P- tubulin. Multiple bands at the top of each gel correspond to different isoforms of OPA1.

[0169] FIG. 2F is a line chart showing relative cell viability over the course of 72 hours after treatment of HEK293 cells with various doses (1 pM, 3 pM, 10 pM, and 20 pM) of either ASO-1 or non-targeting ASO (NT ASO).

[0170] FIG. 2G is a line chart showing relative cell viability 72 hours after treatment of HEK293 cells with various concentrations of cycloheximide (8 ng / ml, 40 ng / ml, 200 ng / ml, 1000 ng / ml, 5000 ng / ml).

[0171] FIGs. 3A-3E represent data from Western Blot analyses showing reduced OPA1 protein levels in OPAl+ / ~ HEK293 cells compared to isogenic OPA1+ / +HEK293 cells. Western Blot gel images are shown in FIG. 3A. Histograms of the quantified Western Blots are shown in FIG. 3B and FIG. 3E, with the latter showing relevant data points. FIG. 3C is a histogram showing relative OPA1 protein expression when isogenic OPA1+ / +HEK293 and OPAl+ / ~ HEK293 cells were treated with Vehicle (no ASO) and ASO-1, and corresponds to quantification of Western Blot data from FIG. 3D. FIG. 3D shows an exemplary Western Blot gel image of OPA1+ / +HEK293 and OPAl+ / ~ HEK293 cells treated with either Vehicle or ASO-1 and probed with antibodies targeting either OPA1 or P-actin. FIG. 3F is a gel image of RT-PCR data from OPA1+ / +HEK293 and OPAl+ / ~ HEK293 cells treated with either DMSO or cycloheximide (CHX) and showing levels of nonproductive (upper bands) and productive (lower bands) OPA1 transcripts. FIG. 3G is a histogram of quantified data from the Western Blot of FIG. 3F showing the percentage of nonproductive transcripts in the tested conditions comprising OPA1+ / +HEK293 and ( / N / ~ HEK293 cells treated with either DMSO or CHX. FIG. 3H is a histogram of qPCR data showing relative levels of nonproductive transcript in the tested conditions comprising OPA1+ / +HEK293 and OPAl+ / ~ HEK293 cells treated with either DMSO or CHX.

[0172] FIGs. 4A-4E show that ASO-1 increases OPA1 mRNA and OPA1 protein expression in OPAl+~ HEK293 cells. FIGs. 4A-4D are histograms of data from OPA1+ / +or OPAl+ / ~ HEK293 cells treated with Vehicle or 10 pM ASO-1 by gymnotic delivery for 72 hours and assessed for effect on OPA1 mRNA expression (FIGs. 4A-4B) and OPA1 protein (FIGs. 4C- 4D) 72 hours post treatment. FIG. 4A is histogram showing data for OPA1 mRNA expression and FIG. 4C is histogram showing data for OPA1 protein levels when OPA1+ / +HEK293 cells are treated with Vehicle or 10 pM ASO-1 by gymnotic delivery. FIG. 4B is histogram showing data for OPA1 mRNA expression and FIG. 4D is histogram showing data for OPA1 protein levels when OPAl+ / ~ HEK293 cells are treated with Vehicle or 10 pM ASO-1 by gymnotic delivery. FIG. 4E is a representative Western Blot image for data in FIGS. 4C-4D.

[0173] FIGs. 5A-5D show that ASO-1 reduces nonproductive exon inclusion and increases OPA1 expression in ADOA patient fibroblast cells. The upper panel of FIG. 5A shows source and identifying information of the wild-type OPA1+ / +fibroblasts tested (WT (ATCC), GM02036, and GM03234), and the lower panel of FIG. 5A contains OPA1 variant information for the three ADOA patients designated as F34, F35 and F36. FIGS. 5B-5C are histograms representing data for WT (ATCC) and all three patient fibroblast cells treated with ASO-1 and harvested for analysis of nonproductive splicing (FIG. 5B) and OPA1 mRNA levels (FIG. 5C). FIG. 5D shows a histogram of OPA1 protein levels determined by Western Blot after transfection and normalized to P-actin. FIGS. 5E-5F are histograms representing data for the three different types of wild-type cells from the upper panel of FIG. 5A (WT (ATCC), GM02036, and GM03234) and the three patient fibroblast cells with different OPA1 pathogenic variants from the lower panel of FIG. 5A that were harvested for analysis of their OPA1 mRNA levels (FIG. 5E) and relative OPA1 protein levels (FIG. 5F) prior to treatment with ASO-1. FIG. 5G is an exemplary Western Blot showing OPA1 protein levels of the three different wild-type fibroblasts and three ADOA patient dermal fibroblasts with different OPA1 pathogenic variants prior to treatment with ASO- 1, and cells were probed with antibodies targeting either OPA1 or P-actin. Multiple bands at the upper part of the gel image of FIG. 5G correspond to different isoforms of OPA1 and the lower bands correspond to P-actin. FIG. 5H shows a histogram of relative nonproductive OPA1 transcript levels when the three wild-type cells (WT (ATCC), GM02036, and GM03234) and three ADOA patient dermal fibroblasts with different OPA1 pathogenic variants (F34, F35, and F36) are treated with either DMSO or CHX. FIG. 51 are exemplary Western Blot gel images showing OPA1 protein levels when wild-type and ADOA patient fibroblasts are treated with either Vehicle (no ASO) or ASO-1, and cells were probed with antibodies targeting either OPA1 or P-actin. Each gel image within FIG. 51 represents a separate experiment, and each lane of each gel represents an individual biological replicate.

[0174] FIGS. 6A-6D are exemplary histograms of oxygen consumption rate that demonstrate ADOA patient-derived fibroblast cells have reduced mitochondrial function. Basal respiration (FIG. 6A), ATP-linked respiration (FIG. 6B), maximal respiration (FIG. 6C) and spare respiratory capacity (FIG. 6D) are shown. FIGS. 6E-6H are exemplary histograms of additional experiments assessing oxygen consumption rate that demonstrate ADOA patient- derived fibroblast cells have reduced mitochondrial function compared to three wild-type cells (WT (ATCC), GM02036, and GM03234). Basal respiration (FIG. 6E), ATP-linked respiration (FIG. 6F), maximal respiration (FIG. 6G) and spare respiratory capacity (FIG. 6H) are shown.

[0175] FIGS. 7A-7D are histograms that show ASO-1 increases mitochondrial function in all three ADOA patients’ fibroblast cells. Oxygen consumption rate measurements were taken from patient fibroblast cells treated with 20, 40 or 60 nM ASO-1 or Vehicle. Basal respiration (FIG. 7A), ATP-linked respiration (FIG. 7B), maximal respiration (FIG. 7C) and spare respiratory capacity (FIG. 7D) are shown. FIGS. 7E-7H are histograms that show the effects of a nontargeting ASO control on mitochondrial function in ADOA patient F35’s fibroblast cells. Oxygen consumption rate measurements were taken from patient fibroblast cells treated with 20, 40 or 60 nM ASO-1 or Vehicle (no ASO). Basal respiration (FIG. 7E), ATP-linked respiration (FIG. 7F), maximal respiration (FIG. 7G) and spare respiratory capacity (FIG. 7H) are shown.

[0176] FIGS. 8A-8D show that surrogate antisense oligonucleotide (ASO) ST-1102 increases OPA1 protein in Wild-Type (WT) rabbit retinal tissue following single intravitreal (IVT) injection. FIG. 8A illustrates the study design for the in vivo rabbit experiment of Example 9. FIG. 8B are histograms representing RT-PCR data measuring nonproductive splicing using rabbit retinal RNA. FIG. 8C shows histograms of quantitated Western Blot data for OPA1 protein normalized to actin. FIG. 8D shows a histogram of ST-1102 detected in retinal tissue on Day 29 by HELISA at all three dose levels.

[0177] FIG. 9 shows RT-PCR gel images from ST-1102-injected rabbit retinas.

[0178] FIG. 10 shows Western Blot images from ST- 1102-injected rabbit retinas.

[0179] FIGS. 11A-11C show histograms demonstrating that intravitreal administration of ASO-1 induces dose-related reduction in nonsense-mediated decay exon inclusion and increased OPA1 protein with sustained tissue exposure in the retina of cynomolgus monkeys. FIG. HA is a histogram of qPCR data measuring the presence of nonproductive exon-containing RNA transcripts in retinal tissue relative to total OPA1 transcripts. FIG. 11B is a histogram representing OPA1 protein quantitated in retinal lysates by ELISA and normalized to total protein in the sample. FIG. 11C is a histogram representing ASO-1 levels in retinal tissue quantitated by HELISA. FIG. HD is a gel image showing RT-PCR data of the right (oculus dexter, OD) and left (oculus sinister, OS) eyes of three cynomolgus monkeys. Upper bands represent nonproductive transcripts andlower bands represent productive transcripts. Numbers at the bottom of the gel refer to the amount of A-X-B relative to the total (A-X-B plus A-B).

[0180] FIG. 12A-12D are images representative of ASO-1 and OPA1 protein in retinal ganglion cells (RGCs) after intravitreal (IVT) administration to cynomolgus monkeys. FIG. 12A shows miRNAscope™ staining with a specific probe to ASO-1 was performed on MDF -fixed, paraffin-embedded cynomolgus monkey eyes to demonstrate cellular location of ASO-1 (red) in RGCs. Increased ASO-1 signal is correlated with increased dose. Nuclei were counterstained with hematoxylin (blue). FIG. 12B shows immunofluorescence staining with OPA1 antibody indicating apparent dose-related protein increase (red) in the RGCs in / near the fovea. Nuclei are stained with DAPI (blue), and magnification was 40X. FIG. 12C shows immunohistochemically (IHC) stained image of RGCs after intravitreal administration of PBS to cynomolgus monkeys. FIG. 12D shows an in situ hybridization (ISH) of RGCs after intravitreal administration of ASO- 1 to cynomolgus monkeys. FIG. 12E is a schematic depicting the experimental procedure entailing the IVT administration of either PBS or ASO-1 to cynomolgus monkeys and evaluation after 4 weeks. The right eyes of cynomolgus monkeys were fixed for in situ hybridization (ISH) and immunohistochemistry (IHC).

[0181] FIG. 13A-13D show that there is dose-related increase in ASO- 1 in retinal ganglion cells (RGCs) after intravitreal (IVT) administration to cynomolgus monkeys. FIGS. 13A-13B are histograms representing quantitated copies of mRNA per cell and percent positive RGCs in or near foveal RGCs. FIGS. 13B-13C are histograms representing quantitated copies of mRNA per cell and percent positive RGCs in peripheral retina.

[0182] FIG. 14A-14B show quantitation of OPA1 protein immunofluorescence in cynomolgus monkey retinal ganglion cells (RGCs) after intravitreal ASO-1 administration. Fluorescence (FL) intensity was quantitated in the RGC layer in foveal / near foveal (FIG. 14A) and peripheral (FIG. 14B) retina.

[0183] FIG. 15A is a schematic depicting the experimental procedure entailing the IVT administration of 50-pl aliquots of either PBS or ASO-1 to cynomolgus monkeys, followed by tissue collection and retina dissection 8 weeks. FIG. 15B shows the treatment conditions of each group in the cynomolgus monkey study. FIG. 15C is a schematic of retina dissection showing a map of the quadrants for downstream evaluation of OPA1 protein expression (QI), degree of ASO- 1 exposure (Q2 and Q4), and OPA1 mRNA expression (Q3). FIG. 15D is a histogram of ASO-1 exposure in the retina for the five dose groups in FIG. 15B as assessed by the measured concentration of ASO-1 (units / g) resulting from hybridization ELISA (hELISA). FIG. 15E is a histogram of NMD exon inclusion in the retina for the five dose groups in FIG. 15B as assessed by the OPA1 NMD exon inclusion relative to the control from qPCR data. FIG. 15F is a histogramof OPA1 protein levels in the retina for the five dose groups in FIG. 15B as assessed by OPA1 measured in units / mg of total protein relative to the control resulting from ELISA.DETAILED DESCRIPTION

[0184] Alternative splicing events in the OPA1 gene can lead to nonproductive mRNA transcripts which in turn can lead to aberrant protein expression, and therapeutic agents which can target the alternative splicing events in the OPA1 gene can modulate the expression level of functional proteins in DS patients and / or inhibit aberrant protein expression. Such therapeutic agents can be used to treat a condition caused by OPA1 protein deficiency.

[0185] One of the alternative splicing events that can lead to nonproductive mRNA transcripts is the inclusion of an extra exon in the mRNA transcript that can induce nonsense- mediated mRNA decay. The present disclosure provides compositions and methods for modulating alternative splicing of OPA1 to increase the production of protein-coding mature mRNA, and thus, translated functional OPA1 protein. These compositions and methods include antisense oligomers (ASOs) that can cause exon skipping, e.g., pseudoexon skipping, and promote constitutive splicing of OPA1 pre-mRNA. In various embodiments, functional OPA1 protein can be increased using the methods of the disclosure to treat a condition caused by OPA1 protein deficiency. mRNA Splicing

[0186] Intervening sequences in RNA sequences or introns are removed by a large and highly dynamic RNA-protein complex termed the spliceosome, which orchestrates complex interactions between primary transcripts, small nuclear RNAs (snRNAs) and a large number of proteins. Spliceosomes assemble ad hoc on each intron in an ordered manner, starting with recognition of the 5’ splice site (5’ss) by U1 snRNA or the 3 ’splice site (3’ss) by the U2 pathway, which involves binding of the U2 auxiliary factor (U2AF) to the 3 ’ ss region to facilitate U2 binding to the branch point sequence (BPS). U2AF is a stable heterodimer composed of a U2AF2-encoded 65-kD subunit (U2AF65), which binds the polypyrimidine tract (PPT), and a U2AF1 -encoded 35- kD subunit (U2AF35), which interacts with highly conserved AG dinucleotides at 3‘ss and stabilizes U2AF65 binding. In addition to the BPS / PPT unit and 3’ss / 5’ss, accurate splicing requires auxiliary sequences or structures that activate or repress splice site recognition, known as intronic or exonic splicing enhancers or silencers. These elements allow genuine splice sites to be recognized among a vast excess of cryptic or pseudo-sites in the genome of higher eukaryotes, which have the same sequences but outnumber authentic sites by an order of magnitude. Although they often have a regulatory function, the exact mechanisms of their activation or repression are poorly understood.

[0187] The decision of whether to splice or not to splice can be typically modeled as a stochastic rather than deterministic process, such that even the most defined splicing signals can sometimes splice incorrectly. However, under normal conditions, pre-mRNA splicing proceeds at surprisingly high fidelity. This is attributed in part to the activity of adjacent cis-acting auxiliary exonic and intronic splicing regulatory elements (ESRs or ISRs). Typically, these functional elements are classified as either exonic or intronic splicing enhancers (ESEs or ISEs) or silencers (ESSs or ISSs) based on their ability to stimulate or inhibit splicing, respectively. Although there is now evidence that some auxiliary cis-acting elements may act by influencing the kinetics of spliceosome assembly, such as the arrangement of the complex between U1 snRNP and the 5’ss, it seems very likely that many elements function in concert with trans-acting RNA-binding proteins (RBPs). For example, the serine- and arginine-rich family of RBPs (SR proteins) is a conserved family of proteins that have a key role in defining exons. SR proteins promote exon recognition by recruiting components of the pre-spliceosome to adjacent splice sites or by antagonizing the effects of ESSs in the vicinity. The repressive effects of ESSs can be mediated by members of the heterogeneous nuclear ribonucleoprotein (hnRNP) family and can alter recruitment of core splicing factors to adjacent splice sites. In addition to their roles in splicing regulation, silencer elements are suggested to have a role in repression of pseudo-exons, sets of decoy intronic splice sites with the typical spacing of an exon but without a functional open reading frame. ESEs and ESSs, in cooperation with their cognate trans-acting RBPs, represent important components in a set of splicing controls that specify how, where and when mRNAs are assembled from their precursors.

[0188] Alternative splicing is a regulated process during gene expression that can result in multiple isoforms of mature mRNA transcripts that are processed from a single primary mRNA transcript that is transcribed from a single gene, and the resultant multiple proteins that are translated from at least some of the multiple mature mRNA isoforms. In this process, particular exons of a gene may be included within or excluded from the final, processed mRNA produced from that gene. Consequently, the proteins translated from alternatively splices mRNAs will contain differences in their amino acid sequence and, in some cases, in their biological functions.

[0189] As described herein, an “alternatively spliced exon” can refer to an exon of a gene that can be either included or excluded naturally from a mature mRNA transcript, thus resulting in different protein products that are translated from the different mature mRNA transcripts. The inclusion or skipping of an alternatively spliced exon can take place naturally in a cell, either randomly, or in a regulated manner, e.g., subject to regulation by external physiological or pathological stimuli, or intracellular signaling. In some cases, the production of alternatively spliced mRNAs, e.g., the splicing of the alternatively spliced exon, is regulated by a system oftrans-acting proteins that bind to cis-acting sites on the primary transcript itself. In some cases, an alternatively spliced exon is a coding exon, e.g., an exon that, when included in the mature mRNA transcript, is translated into an amino acid sequence as part of the protein product translated from the mature mRNA transcript. In some cases, the inclusion of an alternatively spliced exon in the mature mRNA transcript would maintain the canonical open reading frame as compared to a mature mRNA transcript without the alternatively spliced exon, e.g., the number of nucleotides in the alternatively spliced exon is divisible by 3.

[0190] The sequences marking the exon-intron boundaries are degenerate signals of varying strengths that can occur at high frequency within human genes. In multi-exon genes, different pairs of splice sites can be linked together in many different combinations, creating a diverse array of transcripts from a single gene. This is commonly referred to as alternative pre- mRNA splicing. Although most mRNA isoforms produced by alternative splicing can be exported from the nucleus and translated into functional polypeptides, different mRNA isoforms from a single gene can vary greatly in their translation efficiency. Those mRNA isoforms with premature termination codons (PTCs) at least 50 bp upstream of an exon junction complex are likely to be targeted for degradation by the nonsense-mediated mRNA decay (NMD) pathway. Mutations in traditional (BPS / PPT / 3’ss / 5’ss) and auxiliary splicing motifs can cause aberrant splicing, such as exon skipping or cryptic (or pseudo-) exon inclusion or splice-site activation, and contribute significantly to human morbidity and mortality. Both aberrant and alternative splicing patterns can be influenced by natural DNA variants in exons and introns.

[0191] Given that exon-intron boundaries can occur at any of the three positions of a codon, it is clear that only a subset of alternative splicing events can maintain the canonical open reading frame. For example, only exons that are evenly divisible by 3 can be skipped or included in the mRNA without any alteration of reading frame. Splicing events that do not have compatible phases will induce a frame-shift. Unless reversed by downstream events, frame-shifts can certainly lead to one or more PTCs, probably resulting in subsequent degradation by NMD. NMD is a translation-coupled mechanism that eliminates mRNAs containing PTCs. NMD can function as a surveillance pathway that exists in all eukaryotes. NMD can reduce errors in gene expression by eliminating mRNA transcripts that contain premature stop codons. Translation of these aberrant mRNAs could, in some cases, lead to deleterious gain-of-function or dominant-negative activity of the resulting proteins. NMD targets not only transcripts with PTCs but also a broad array of mRNA isoforms expressed from many endogenous genes, suggesting that NMD is a master regulator that drives both fine and coarse adjustments in steady-state RNA levels in the cell.

[0192] An NMD-inducing exon (“NIE” or “NMD exon”) is an exon or a pseudo-exon that is a region within an intron and can activate the NMD pathway if included in a mature RNAtranscript. In constitutive splicing events, the intron containing an NMD exon is usually spliced out, but the intron or a portion thereof (e.g., NMD exon) may be retained during alternative or aberrant splicing events. Mature mRNA transcripts containing such an NMD exon may be nonproductive due to frame shifts which induce the NMD pathway. Inclusion of a NMD exon in mature RNA transcripts may downregulate gene expression. mRNA transcripts containing an NMD exon may be referred to as “NIE-containing mRNA” or “NMD exon mRNA” in the current disclosure.

[0193] Cryptic (or pseudo- splice sites) have the same splicing recognition sequences as genuine splice sites but are not used in splicing reactions. They outnumber genuine splice sites in the human genome by an order of a magnitude and are normally repressed by thus far poorly understood molecular mechanisms. Cryptic 5’ splice sites have the consensus NNN / GUNNNN or NNN / GCNNNN where N is any nucleotide and / is the exon-intron boundary. Cryptic 3’ splice sites have the consensus NAG / N. Their activation is positively influenced by surrounding nucleotides that make them more similar to the optimal consensus of authentic splice sites, namely MAG / GURAGU and YAG / G, respectively, where M is C or A, R is G or A, and Y is C or U.

[0194] Splice sites and their regulatory sequences can be readily identified by a skilled person using suitable algorithms publicly available, listed for example, in Kralovicova, J. and Vorechovsky, I. (2007) Global control of aberrant splice site activation by auxiliary splicing sequences: evidence for a gradient in exon and intron definition. Nucleic Acids Res., 35, 6399- 6413 (www.ncbi.nlm.nih.gov / pmc / articles / PMC2095810 / pdf / gkm680. pdf).

[0195] The cryptic splice sites or splicing regulatory sequences may compete for RNA- binding proteins, such as U2AF, with a splice site of the NMD exon. In some embodiments, an agent may bind to a cryptic splice site or splicing regulatory sequence to prevent binding of RNA- binding proteins and thereby favor binding of RNA-binding proteins to the NMD exon splice sites.

[0196] In some embodiments, the cryptic splice site may not comprise the 5’ or 3’ splice site of the NMD exon. In some embodiments, the cryptic splice site may be at least 10 nucleotides, at least 20 nucleotides, at least 50 nucleotides, at least 100 nucleotides or at least 200 nucleotides upstream of the NMD exon 5’ splice site. In some embodiments, the cryptic splice site may be at least 10 nucleotides, at least 20 nucleotides, at least 50 nucleotides, at least 100 nucleotides, at least 200 nucleotides downstream of the NMD exon 3’ splice site.Target Transcripts

[0197] In some embodiments, the methods and compositions of the present disclosure exploit the presence of NMD exon in the pre-mRNA transcribed from the OPA1 gene. Splicing of the identified OPA1 NMD exon pre-mRNA species to produce functional mature OPA1 mRNA may be induced using an agent such as an ASO that stimulates exon skipping of an NMD exon.Induction of exon skipping may result in inhibition of an NMD pathway. The resulting mature 0PA1 mRNA can be translated normally without activating NMD pathway, thereby increasing the amount of OPA1 protein in the patient’s cells and alleviating symptoms of a condition or disease associated with 0PA1 deficiency, such as an eye disease or condition, Optic atrophy type 1, autosomal dominant optic atrophy (ADOA), ADOA-plus syndrome; a mitochondrial disorder; glaucoma; normal tension glaucoma; Charcot-Marie-Tooth disease; mitochondria dysfunction; diabetic retinopathy; age-related macular degeneration; retinal ganglion cell death; mitochondrial fission-mediated mitochondrial dysfunction; progressive external ophthalmoplegia; deafness; ataxia; motor neuropathy; sensory neuropathy; myopathy; Behr syndrome; brain dysfunction; encephalopathy; peripheral neuropathy; fatal infantile mitochondrial encephalomyopathy; hypertrophic cardiomyopathy; spastic ataxic syndrome; sensory motor peripheral neuropathy; hypotonia; gastrointestinal dysmotility and dysphagia; optic atrophy; optic atrophy plus syndrome; Mitochondrial DNA depletion syndrome 14; late-onset cardiomyopathy; diabetic cardiomyopathy; Alzheimer’s Disease; focal segmental glomerulosclerosis; kidney disease; Huntington’s Disease; cognitive function decline in healthy aging; Prion diseases; late onset dementia and parkinsonism; mitochondrial myopathy; Leigh syndrome; Friedreich’s ataxia; Parkinson’s disease; MELAS (Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes); pyruvate dehydrogenase complex deficiency; chronic kidney disease; Leber’s hereditary optic neuropathy; obesity; age-related systemic neurodegeneration; skeletal muscle atrophy; heart and brain ischemic damage; or massive liver apoptosis.

[0198] In some embodiments, the methods and compositions of the present disclosure exploit the alternative splicing of the pre-mRNA transcribed from the OP Al gene. In some cases, splicing of a coding exon, e.g, an alternatively spliced exon, e.g, OPA1 exon 7 (or an exon encoded by genomic region spanning from GRCh38 / hg38: chr3 193626092 to 193626202), can modulate the level of OPA1 protein expressed from the OPA1 gene. As described herein, the term “OPA1 exon 7” or grammatically equivalents thereof, is used interchangeably with the term “exon (GRCh38 / hg38: chr3 193626092 to 193626202)” or “an exon encoded by genomic region spanning from GRCh38 / hg38: chr3 193626092 to 193626202.” Without wishing to be bound by a certain theory, the presence or absence of an amino acid sequence encoded by exon 7 or exon (GRCh38 / hg38: chr3 193626092 to 193626202) can modulate the stability of the OPA1 protein. For instance, in some cases, the OPA1 protein encoded by a mature mRNA transcript that lacks exon 7 can have fewer proteolytic cleavage sites as compared to an OPA1 protein encoded by a corresponding mature mRNA transcript that has contains exon 7. In some cases, the OP Al protein an OPA1 protein encoded by a corresponding mature mRNA transcript that has contains encoded by a mature mRNA transcript that lacks exon 7 is a functional protein. The OPA1 protein encodedby a mature mRNA transcript that lacks exon 7 can be at least partially functional as compared to an OP Al protein encoded by a corresponding mature mRNA transcript that has contains exon 7. In some cases, the OPA1 protein encoded by a mature mRNA transcript that lacks exon 7 is at least partially functional as compared to a full-length wild-type OPA1 protein. In some cases, increase of OPA1 protein encoded by a mature mRNA transcript that lacks exon 7 in a cell can result in more functional OPA1 protein in the cell, due to the higher stability of the OPA1 protein lacking exon 7 and its at least partial functional equivalence.

[0199] In other embodiments, a coding exon of OPA1 pre-mRNA other than exon 7 is targeted by an agent disclosed herein, which promotes exclusion of the coding exon other than exon 7. In these other embodiments, the agent that promotes exclusion of the coding exon other than exon 7 increases expression of OPA1 protein encoded by a mature mRNA transcript that lacks the excluded exon.

[0200] Alternative splicing of the OPA1 pre-mRNA species, e.g., skipping of a coding exon, e.g., an alternatively spliced exon, e.g., exon 7, to produce functional mature OPA1 protein may be induced using an agent such as an ASO that stimulates the exon skipping. Induction of exon skipping may result in modulation of levels of different alternatively spliced mRNA transcripts. The resulting mature OPA1 mRNA can be translated into different OPA1 proteins, thereby modulating the amount of OPA1 protein in the patient’s cells and alleviating symptoms of a condition or disease associated with OPA1 deficiency, such as an eye disease or condition, Optic atrophy type 1, autosomal dominant optic atrophy (ADO A), ADOA-plus syndrome; a mitochondrial disorder; glaucoma; normal tension glaucoma; Charcot-Mari e-tooth disease; mitochondria dysfunction; diabetic retinopathy; age-related macular degeneration; retinal ganglion cell death; mitochondrial fission-mediated mitochondrial dysfunction; progressive external ophthalmoplegia; deafness; ataxia; motor neuropathy; sensory neuropathy; myopathy; Behr syndrome; brain dysfunction; encephalopathy; peripheral neuropathy; fatal infantile mitochondrial encephalomyopathy; hypertrophic cardiomyopathy; spastic ataxic syndrome; sensory motor peripheral neuropathy; hypotonia; gastrointestinal dysmotility and dysphagia; optic atrophy; optic atrophy plus syndrome; Mitochondrial DNA depletion syndrome 14; late-onset cardiomyopathy; diabetic cardiomyopathy; Alzheimer’s Disease; focal segmental glomerulosclerosis; kidney disease; Huntington’s Disease; cognitive function decline in healthy aging; Prion diseases; late onset dementia and parkinsonism; mitochondrial myopathy; Leigh syndrome; Friedreich’s ataxia; Parkinson’s disease; MELAS (Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes); pyruvate dehydrogenase complex deficiency; chronic kidney disease; Leber’s hereditary optic neuropathy; obesity; age-related systemicneurodegeneration; skeletal muscle atrophy; heart and brain ischemic damage; or massive liver apoptosis.

[0201] In some embodiments, the diseases or conditions that can be treated or ameliorated using the method or composition disclosed herein are not directly associated with the target protein (gene) that the therapeutic agent targets. In some embodiments, a therapeutic agent provided herein can target a protein (gene) that is not directly associated with a disease or condition, but the modulation of expression of the target protein (gene) can treat or ameliorate the disease or condition.

[0202] In some embodiments, the antisense oligomer modulates splicing of a nonsense- mediated RNA decay-inducing exon (NMD exon) from a pre-mRNA in a cell of the subject, wherein the pre-mRNA encodes the OP Al protein and comprises the NMD exon, thereby modulating a level of processed mRNA that is processed from the pre-mRNA, and modulating expression of the OPA1 protein in the cell.

[0203] In some embodiments, the antisense oligomer: (a) binds to a targeted portion of the pre-mRNA; (b) modulates binding of a factor involved in splicing of the NMD exon; or (c) a combination of (a) and (b).

[0204] In some embodiments, the targeted portion of the pre-mRNA is proximal to the NMD exon.

[0205] In various embodiments, the present disclosure provides a therapeutic agent which can target OPA1 mRNA transcripts to modulate splicing or protein expression level. The therapeutic agent can be a small molecule, polynucleotide, or polypeptide. In some embodiments, the therapeutic agent is an ASO. Various regions or sequences on the OPA1 pre-mRNA can be targeted by a therapeutic agent, such as an ASO. In some embodiments, the ASO targets an OPA1 pre-mRNA transcript containing an NMD exon. In some embodiments, the ASO targets a sequence within an NMD exon of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence upstream (or 5’) from the 5’ end of an NMD exon (3’ss) of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence downstream (or 3’) from the 3’ end of an NMD exon (5 ’ ss) of an OP Al pre-mRNA transcript. In some embodiments, the ASO targets a sequence that is within an intron flanking on the 5’ end of the NMD exon of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence that is within an intron flanking the 3 ’ end of the NMD exon of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence comprising an NMD exon-intron boundary of an OPA1 pre-mRNA transcript. An NMD exon-intron boundary can refer to the junction of an intron sequence and an NMD exon region. The intron sequence can flank the 5’ end of the NMD exon, or the 3’ end of the NMD exon. In some embodiments, the ASO targets a sequence within an exon of an OPA1 pre-mRNAtranscript. In some embodiments, the ASO targets a sequence within an intron of an OPA1 pre- mRNA transcript. In some embodiments, the ASO targets a sequence comprising both a portion of an intron and a portion of an exon of an OPA1 pre-mRNA transcript.

[0206] In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides upstream (or 5’) from the 5’ end of the NMD exon. In some embodiments, the ASO targets a sequence about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, or about 250 to about 300 nucleotides upstream (or 5’) from the 5’ end of the NMD exon region. In some embodiments, the ASO may target a sequence more than 300 nucleotides upstream from the 5’ end of the NMD exon. In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides downstream (or 3’) from the 3’ end of the NMD exon. In some embodiments, the ASO targets a sequence about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, or about 250 to about 300 nucleotides downstream from the 3’ end of the NMD exon. In some embodiments, the ASO targets a sequence more than 300 nucleotides downstream from the 3’ end of the NMD exon.

[0207] In some embodiments, the OPA1 NMD exon-containing pre-mRNA transcript is encoded by a genetic sequence with at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1. In some embodiments, the OPA1 NMD exon pre- mRNA transcript comprises a sequence with at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of SEQ ID NOs: 2-5.

[0208] In some embodiments, the OPA1 NMD exon-containing pre-mRNA transcript (or NMD exon mRNA) comprises a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 2-5. In some embodiments, OP Al NMD exoncontaining pre-mRNA transcript (or NMD exon mRNA) is encoded by a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 2-5. In some embodiments, the targeted portion of the NMD exon mRNA comprises a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5.

[0209] In some embodiments, the ASO targets exon 6x of an OPA1 NMD exon-containing pre-mRNA comprising NIE exon 6, exon 7x of an OP Al NMD exon-containing pre-mRNA comprising NIE exon 7, or exon 28x of an OPA1 NMD exon-containing pre-mRNA comprising NIE exon 28. In some embodiments, the ASO targets exon (GRCh38 / hg38: chr3 193628509 193628616) of OPA1 pre-mRNA; or exon (GRCh38 / hg38: chr3 193603500 193603557) ofOPAl. In some embodiments, the ASO targets an NMD exon of OPA1 pre-mRNA other than NMD exon (GRCh38 / hg38: chr3 193628509 193628616).

[0210] In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from the 5’ end of exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl . In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from GRCh38 / hg38: chr3 193628509 of OPAP, or GRCh38 / hg38: chr3 193603500 of OPAl.

[0211] In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from the 5’ end of exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl. In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from GRCh38 / hg38: chr3 193628509 of OPAl,' or GRCh38 / hg38: chr3 193603500 of OPAl.

[0212] In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from the 3’ end of exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl. In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from GRCh38 / hg38: chr3 193628616 of OPAl,' or GRCh38 / hg38: chr3 193603557 of OPAl.

[0213] In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from the 3’ end of exon 6x of OPA L exon 7x of OPA L or exon 28x of OPAl. In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from GRCh38 / hg38: chr3 193628616 of OPAl,' or GRCh38 / hg38: chr3 193603557 of OPAL

[0214] In some embodiments, the ASO has a sequence complementary to the targeted portion of the NMD exon mRNA according to any one of SEQ ID NOs: 2-5, or 279.

[0215] In some embodiments, the ASO targets a sequence upstream from the 5’ end of an NMD exon. For example, ASOs targeting a sequence upstream from the 5’ end of an NMD exon (exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl comprises a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complimentary to at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3. For example, ASOs targeting a sequence upstream from the 5’ end of an NMD exon (e.g., exon (GRCh38 / hg38: chr3 193628509 to 193628616) of OPAL, or exon (GRCh38 / hg38: chr3 193603500 193603557) of OPAl can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 4 or 5.

[0216] In some embodiments, the ASOs target a sequence containing an exon-intron boundary (or junction). For example, ASOs targeting a sequence containing an exon-intron boundary can comprise a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complimentary to at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5. In some embodiments, the ASOs target a sequence downstream from the 3’ end of an NMD exon. For example, ASOs targeting a sequence downstream from the 3’ end of an NMD exon (e.g., exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl} can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 2 or 3, or at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3. For example, ASOs targeting a sequence downstream from the 3’ end of an NMD exon (e.g., exon (GRCh38 / hg38: chr3 193628509 to 193628616) of OPAL, or exon (GRCh38 / hg38: chr3 193603500 to 193603557) of OPAl} can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 4 or 5, or at least 8 contiguous nucleic acids of SEQ ID NO: 4 or 5. In some embodiments, ASOs target a sequence within an NMD exon.

[0217] In some embodiments, the ASO targets exon 6x of an OP Al NMD exon-containing pre-mRNA comprising NIE exon 6, exon 7x of an OP Al NMD exon-containing pre-mRNA comprising NIE exon 7, or exon 28x of an OPA1 NMD exon-containing pre-mRNA comprising NIE exon 28. In some embodiments, the ASO targets a sequence downstream (or 3’) from the 5’ end of exon 6x, exon 7x, or exon 28x of an OP Al pre-mRNA. In some embodiments, the ASO targets a sequence upstream (or 5’) from the 3’ end of exon 6x, exon 7x, or exon 28x of an OPA1 pre-mRNA.

[0218] In some embodiments, the targeted portion of the OPA1 NMD exon-containing pre- mRNA is in intron 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. In some embodiments, hybridization of an ASO to the targeted portion of the NMD exon pre- mRNA results in exon skipping of at least one of NMD exon within intron 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, and subsequently increases OPA1 protein production. In some embodiments, the targeted portion of the OPA1 NMD exon-containing pre-mRNA is in intron 6 of OPA1, or intron 28 of OPAl. In some embodiments, the targeted portion of the OPAl NMD exon-containing pre-mRNA is intron (GRCh38 / hg38: chr3 193626203 to 193631611) of OPAP, or intron (GRCh38 / hg38: chr3 193593374 to 193614710) of OPAl.

[0219] In some embodiments, the methods and compositions of the present disclosure are used to increase the expression of OPAl by inducing exon skipping of a pseudo-exon of an OPAl NMD exon-containing pre-mRNA. In some embodiments, the pseudo-exon is a sequence within any of introns 1-50. In some embodiments, the pseudo-exon is a sequence within any of introns 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33,34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50. In some embodiments, the pseudo-exon can be an OPAl intron or a portion thereof. In some embodiments, the pseudoexon is within intron 6 of OPAl, or intron 28 of OPAl. In some embodiments, the pseudo-exon is within intron (GRCh38 / hg38: chr3 193626203 to 193631611) of OP A 7; orintron (GRCh38 / hg38: chr3 193593374 to 193614710) of OPAl.

[0220] In some embodiments, the ASO targets an OPAl pre-mRNA transcript to induce exon skipping of a coding exon, e.g., an alternatively spliced exon. In some embodiments, the ASO targets a sequence within a coding exon, e.g., an alternatively spliced exon, of an OPAl pre- mRNA transcript. In some embodiments, the ASO targets a sequence upstream (or 5’) from the 5’ end of a coding exon (3’ss) of an OPAl pre-mRNA transcript. In some embodiments, the ASO targets a sequence downstream (or 3’) from the 3’ end of a coding exon (5’ss) of an OPAl pre- mRNA transcript. In some embodiments, the ASO targets a sequence that is within an intronflanking on the 5’ end of the coding exon of an OP Al pre-mRNA transcript. In some embodiments, the ASO targets a sequence that is within an intron flanking the 3’ end of the coding exon of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence comprising an exon-intron boundary of an OPA1 pre-mRNA transcript. An exon-intron boundary can refer to the junction of an intron sequence and an exon sequence. The intron sequence can flank the 5’ end of the coding exon, or the 3’ end of the coding exon. In some embodiments, the ASO targets a sequence within an exon of an OP Al pre-mRNA transcript. In some embodiments, the ASO targets a sequence within an intron of an OPA1 pre-mRNA transcript. In some embodiments, the ASO targets a sequence comprising both a portion of an intron and a portion of an exon of an OPA1 pre-mRNA transcript.

[0221] In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides upstream (or 5’) from the 5’ end of the coding exon, e.g., alternatively spliced exon. In some embodiments, the ASO targets a sequence about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, or about 250 to about 300 nucleotides upstream (or 5’) from the 5’ end of the coding exon region. In some embodiments, the ASO may target a sequence more than 300 nucleotides upstream from the 5’ end of the coding exon. In some embodiments, the ASO targets a sequence about 4 to about 300 nucleotides downstream (or 3’) from the 3’ end of the coding exon. In some embodiments, the ASO targets a sequence about 1 to about 20 nucleotides, about 20 to about 50 nucleotides, about 50 to about 100 nucleotides, about 100 to about 150 nucleotides, about 150 to about 200 nucleotides, about 200 to about 250 nucleotides, or about 250 to about 300 nucleotides downstream from the 3’ end of the coding exon. In some embodiments, the ASO targets a sequence more than 300 nucleotides downstream from the 3’ end of the coding exon.

[0222] In some embodiments, the OPA1 pre-mRNA transcript is encoded by a genetic sequence with at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 1. In some embodiments, the OPA1 pre-mRNA transcript comprises a sequence with at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to any one of SEQ ID NOs: 2-5.

[0223] In some embodiments, the OPA1 pre-mRNA transcript (or NMD exon mRNA) comprises a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 2-5. In some embodiments, OPA1 pre-mRNA transcript (or NMD exon mRNA) is encoded by a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to any one of SEQ ID NOs: 2-5. In some embodiments, the targeted portion of the OP Al pre-mRNA comprises a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100%sequence identity to a region comprising at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5.

[0224] In some embodiments, the ASO targets exon 7 of an OPA1 pre-mRNA, i.e., the ASO targets exon (GRCh38 / hg38: chr3 193626092 to 193626202) of OPA1 pre-mRNA.

[0225] In some embodiments, the ASO targets a coding exon of an OP Al pre-mRNA other than exon 7, i.e., the ASO targets an exon of OPA1 pre-mRNA other than exon defined by (GRCh38 / hg38: chr3 193626092 to 193626202).

[0226] In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from the 5’ end of exon 7 of OPA1. In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from GRCh38 / hg38: chr3 193626092 of OPAL

[0227] In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from the 5’ end of exon 7 of OPAL In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream (or 5’) from GRCh38 / hg38: 193626092 of OPAL

[0228] In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from the 3’ end of exon 7 of OPAL In some embodiments, the ASO targets a sequence about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from GRCh38 / hg38: chr3 193626202 of OPA 1.

[0229] In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from the 3’ end of exon 7 of OPA1. In some embodiments, the ASO targets a sequence at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream (or 3’) from GRCh38 / hg38: chr3 193626202 of OPAP

[0230] In some embodiments, the ASO has a sequence complementary to the targeted portion of the NMD exon mRNA according to any one of SEQ ID NOs: 2-5, or 277.

[0231] In some embodiments, the ASO targets a sequence upstream from the 5’ end of a coding exon, e.g., an alternatively spliced exon. For example, ASOs targeting a sequence upstream from the 5’ end of a coding exon (e.g., exon 7 of OPA ) comprises a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complimentary to at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3. For example, ASOs targeting a sequence upstream from the 5’ end of a coding exon (e.g., exon (GRCh38 / hg38: 193626092 to 193626202) of OPAP) can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 4 or 5.

[0232] In some embodiments, the ASOs target a sequence containing an exon-intron boundary (or junction). For example, ASOs targeting a sequence containing an exon-intron boundary can comprise a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complimentary to at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5. In some embodiments, the ASOs target a sequence downstream from the 3’ end of a coding exon, e.g., an alternatively spliced exon. For example, ASOs targeting a sequence downstream from the 3’ end of a coding exon (e.g., exon 7 of OPAP) can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 2 or 3, or at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3. For example, ASOs targeting a sequence downstream from the 3’ end of a coding exon (e.g., exon 7 of OPAP) can comprise a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 4 or 5, or at least 8 contiguous nucleic acids of SEQ ID NO: 4 or 5. In some embodiments, ASOs target a sequence within a coding exon, e.g., an alternatively spliced exon.Protein Expression

[0233] In some embodiments, the methods described herein are used to increase the production of a functional OPA1 protein or RNA. As used herein, the term “functional” refers to the amount of activity or function of an OPA1 protein or RNA that is necessary to eliminate any one or more symptoms of a treated condition or disease, e.g., Optic atrophy type 1. In some embodiments, the methods are used to increase the production of a partially functional OPA1 protein or RNA. As used herein, the term “partially functional” refers to any amount of activity or function of the OPA1 protein or RNA that is less than the amount of activity or function that is necessary to eliminate or prevent any one or more symptoms of a disease or condition. In some embodiments, a partially functional protein or RNA will have at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% less activity relative to the fully functional protein or RNA.

[0234] In some embodiments, the method is a method of increasing the expression of the OP Al, protein by cells of a subject having an OP Al pre-mRNA, wherein the subject has a disease or condition, e.g., Optic atrophy type 1, caused by a deficient amount of activity of OPA1 protein, and wherein the deficient amount of the OPA1 protein is caused by haploinsufficiency of the OPA1 protein. In such an embodiment, the subject has a first allele encoding a functional OPA1 protein, and a second allele from which the OPA1 protein is not produced. In another such embodiment, the subject has a first allele encoding a functional OPA1 protein, and a second allele encoding a nonfunctional OPA1 protein. In another such embodiment, the subject has a first allele encoding a functional OPA1 protein, and a second allele encoding a partially functional OPA1 protein. In any of these embodiments, the antisense oligomer binds to a targeted portion of the OPA1 pre- mRNA transcribed from the second allele, thereby inducing exon skipping of the pseudo-exon from the pre-mRNA, and causing an increase in the level of mature mRNA encoding functional OPA1 protein, and an increase in the expression of the OPA1 protein in the cells of the subject.

[0235] In some embodiments, the method is a method of increasing the expression of the OPA1 protein by cells of a subject having an OP Al pre-mRNA, wherein the subject has a disease or condition caused by a deficient amount of activity of OPA1 protein, and wherein the deficient amount of the OPA1 protein is caused by autosomal recessive inheritance.

[0236] In some embodiments, the method is a method of increasing the expression of the OPA1 protein by cells of a subject having an OP Al pre-mRNA, wherein the subject has a disease or condition, e.g., Optic atrophy type 1, caused by a deficient amount of activity of OPA1, protein, and wherein the deficient amount of the OPA1 protein is caused by autosomal dominant inheritance.

[0237] In related embodiments, the method is a method of using an ASO to increase the expression of a protein or functional RNA. In some embodiments, an ASO may be used to increase the expression of OPA1 protein in cells of a subject having an OPA1 pre-mRNA, wherein the subject has a deficiency, e.g., Optic atrophy type 1; in the amount or function of an OPA1 protein.

[0238] In some embodiments, the pre-mRNA transcript that encodes the protein that is causative of the disease or condition is targeted by the agent, e.g., the oligonucleotides, described herein. In some cases, it is the NMD exon-containing pre-mRNA transcript targeted by the agent, e.g., the oligonucleotides, described herein. In some cases, the agent, e.g., the oligonucleotides, described herein, are designed to target a coding exon of the pre-mRNA. In some cases, the agent, e.g., the oligonucleotides, described herein can induce skipping of the NMD exon, a coding exon, or both. In some embodiments, a NMD exon-containing pre-mRNA transcript that encodes a protein that is not causative of the disease is targeted by the ASOs. For example, a disease that is the result of a mutation or deficiency of a first protein in a particular pathway may be ameliorated by targeting a pre-mRNA that encodes a second protein, thereby increasing production of the second protein. In some embodiments, the function of the second protein is able to compensate for the mutation or deficiency of the first protein (which is causative of the disease or condition).

[0239] In some embodiments, the subject has:(a) a first mutant allele from which(i) the OPA1 protein is produced at a reduced level compared to production from a wildtype allele,(ii) the OPA1 protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the OPA1 protein or functional RNA is not produced; and(b) a second mutant allele from which(i) the OPA1 protein is produced at a reduced level compared to production from a wildtype allele,(ii) the OPA1 protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the OP Al protein is not produced, and wherein the NMD exon-containing pre-mRNA is transcribed from the first allele and / or the second allele. In these embodiments, the ASO binds to a targeted portion of the NMD exon-containing pre-mRNA transcribed from the first allele or the second allele, thereby inducing exon skipping of the pseudo-exon from the NMD exon-containing pre-mRNA, and causing an increase in the level of mRNA encoding OPA1 protein and an increase in the expression of the target protein or functional RNA in the cells of the subject. In theseembodiments, the target protein or functional RNA having an increase in expression level resulting from the exon skipping of the pseudo-exon from the NMD exon-containing pre- mRNA may be either in a form having reduced function compared to the equivalent wildtype protein (partially-functional), or having full function compared to the equivalent wild-type protein (fully-functional).

[0240] In some embodiments, the subject has:(a) a first mutant allele from which(i) the OPA1 protein is produced at a reduced level compared to production from a wildtype allele,(ii) the OPA1 protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the OPA1 protein or functional RNA is not produced; and(b) a second mutant allele from which(i) the OPA1 protein is produced at a reduced level compared to production from a wildtype allele,(ii) the OPA1 protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the OP Al protein is not produced, and wherein the OPA1 pre-mRNA is transcribed from the first allele and / or the second allele. In these embodiments, the ASO binds to a targeted portion of the OPA1 pre-mRNA transcribed from the first allele or the second allele, thereby inducing exon skipping of a coding exon from the OP Al pre-mRNA, and causing an increase in the expression of the target OPA1 protein in the cells of the subject. In these embodiments, the target OPA1 protein having an increase in expression level resulting from the exon skipping of the coding exon from the OP Al pre-mRNA may be either in a form having reduced function compared to the equivalent full-length wild-type protein (partially-functional), or having full function compared to the equivalent full-length wild-type protein (fully-functional).

[0241] In some embodiments, the level of mRNA encoding OPA1 protein is increased 1.1 to 10-fold, when compared to the amount of mRNA encoding OPA1 protein that is produced in a control cell, e.g., one that is not treated with the antisense oligomer or one that is treated with an antisense oligomer that does not bind to the targeted portion of the OPA1 pre-mRNA.

[0242] In some embodiments, a subject treated using the methods of the present disclosure expresses a partially functional OPA1 protein from one allele, wherein the partially functional OPA1 protein may be caused by a frameshift mutation, a nonsense mutation, a missense mutation, or a partial gene deletion. In some embodiments, a subject treated using the methods of thedisclosure expresses a nonfunctional OPA1 protein from one allele, wherein the nonfunctional OPA1 protein may be caused by a frameshift mutation, a nonsense mutation, a missense mutation, a partial gene deletion, in one allele. In some embodiments, a subject treated using the methods of the disclosure has an OPA1 whole gene deletion, in one allele.Exon Inclusion

[0243] As used herein, a “NMD exon-containing pre-mRNA” is a pre-mRNA transcript that contains at least one pseudo-exon. Alternative or aberrant splicing can result in inclusion of the at least one pseudo-exon in the mature mRNA transcripts. The terms “mature mRNA,” and “fully-spliced mRNA,” are used interchangeably herein to describe a fully processed mRNA. Inclusion of the at least one pseudo-exon can be nonproductive mRNA and lead to NMD of the mature mRNA. NMD exon-containing mature mRNA may sometimes lead to aberrant protein expression.

[0244] In some embodiments, the included pseudo-exon is the most abundant pseudo-exon in a population of NMD exon-containing pre-mRNAs transcribed from the gene encoding the target protein in a cell. In some embodiments, the included pseudo-exon is the most abundant pseudo-exon in a population of NMD exon-containing pre-mRNAs transcribed from the gene encoding the target protein in a cell, wherein the population of NMD exon-containing pre-mRNAs comprises two or more included pseudo-exons. In some embodiments, an antisense oligomer targeted to the most abundant pseudo-exon in the population of NMD exon-containing pre- mRNAs encoding the target protein induces exon skipping of one or two or more pseudo-exons in the population, including the pseudo-exon to which the antisense oligomer is targeted or binds. In some embodiments, the targeted region is in a pseudo-exon that is the most abundant pseudo-exon in a NMD exon-containing pre-mRNA encoding the OPA1 protein.

[0245] The degree of exon inclusion can be expressed as percent exon inclusion, e.g., the percentage of transcripts in which a given pseudo-exon is included. In brief, percent exon inclusion can be calculated as the percentage of the amount of RNA transcripts with the exon inclusion, over the sum of the average of the amount of RNA transcripts with exon inclusion plus the average of the amount of RNA transcripts with exon exclusion.

[0246] In some embodiments, an included pseudo-exon is an exon that is identified as an included pseudo-exon based on a determination of at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50%, inclusion. In embodiments, a included pseudo-exon is an exon that is identified as a included pseudo-exon based on a determination of about 5% to about 100%, about 5% to about 95%, about 5% to about 90%, about 5% to about 85%, about 5% to about 80%, about 5% to about 75%, about 5% to about 70%, about 5% to about65%, about 5% to about 60%, about 5% to about 55%, about 5% to about 50%, about 5% to about 45%, about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 5% to about 20%, about 5% to about 15%, about 10% to about 100%, about 10% to about 95%, about 10% to about 90%, about 10% to about 85%, about 10% to about 80%, about 10% to about 75%, about 10% to about 70%, about 10% to about 65%, about 10% to about 60%, about 10% to about 55%, about 10% to about 50%, about 10% to about 45%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 25%, about 10% to about 20%, about 15% to about 100%, about 15% to about 95%, about 15% to about 90%, about 15% to about 85%, about 15% to about 80%, about 15% to about 75%, about 15% to about 70%, about 15% to about 65%, about 15% to about 60%, about 15% to about 55%, about 15% to about 50%, about 15% to about 45%, about 15% to about 40%, about 15% to about 35%, about 15% to about 30%, about 15% to about 25%, about 20% to about 100%, about 20% to about 95%, about 20% to about 90%, about 20% to about 85%, about 20% to about 80%, about 20% to about 75%, about 20% to about 70%, about 20% to about 65%, about 20% to about 60%, about 20% to about 55%, about 20% to about 50%, about 20% to about 45%, about 20% to about 40%, about 20% to about 35%, about 20% to about 30%, about 25% to about 100%, about 25% to about 95%, about 25% to about 90%, about 25% to about 85%, about 25% to about 80%, about 25% to about 75%, about 25% to about 70%, about 25% to about 65%, about 25% to about 60%, about 25% to about 55%, about 25% to about 50%, about 25% to about 45%, about 25% to about 40%, or about 25% to about 35%, inclusion. ENCODE data (described by, e.g., Tilgner, etal., 2012, “Deep sequencing of subcellular RNA fractions shows splicing to be predominantly co-transcriptional in the human genome but inefficient for IncRNAs,” Genome Research 22(9): 1616-25) can be used to aid in identifying exon inclusion.

[0247] In some embodiments, contacting cells with an ASO that is complementary to a targeted portion of an OPA1 pre-mRNA transcript results in an increase in the amount of OPA1 protein produced by at least 10, 20, 30, 40, 50, 60, 80, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 1000%, compared to the amount of the protein produced by a cell in the absence of the ASO / absence of treatment. In some embodiments, the total amount of OPA1 protein produced by the cell to which the antisense oligomer is contacted is increased about 20% to about 300%, about 50% to about 300%, about 100% to about 300%, about 150% to about 300%, about 20% to about 50%, about 20% to about 100%, about 20% to about 150%, about 20% to about 200%, about 20% to about 250%, about 50% to about 100%, about 50% to about 150%, about 50% to about 200%, about 50% to about 250%, about 100% to about 150%, about 100% to about 200%, about 100% to about 250%, about 150% to about 200%, about 150% to about 250%, about 200% to about 250%, at least about 10%, at least about 20%, at least about 50%, at least about 100%, at leastabout 150%, at least about 200%, at least about 250%, or at least about 300%, compared to the amount of target protein produced by a control compound. In some embodiments, the total amount of OPA1 protein produced by the cell to which the antisense oligomer is contacted is increased about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1 -fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5- fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to the amount of target protein produced by a control compound. A control compound can be, for example, an oligonucleotide that is not complementary to a targeted portion of the pre-mRNA.

[0248] In some embodiments, contacting cells with an ASO that is complementary to a targeted portion of an OPA1 pre-mRNA transcript results in an increase in the amount of mRNA encoding OPA1, including the mature mRNA encoding the target protein. In some embodiments, the amount of mRNA encoding OP Al protein, or the mature mRNA encoding the OP Al protein, is increased by at least 10, 20, 30, 40, 50, 60, 80, 100, 150, 200, 250, 300, 350, 400, 450, 500, or 1000%, compared to the amount of the protein produced by a cell in the absence of the ASO / absence of treatment. In some embodiments, the total amount of the mRNA encoding OPA1 protein, or the mature mRNA encoding OPA1 protein produced in the cell to which the antisense oligomer is contacted is increased about 20% to about 300%, about 50% to about 300%, about 100% to about 300%, about 150% to about 300%, about 20% to about 50%, about 20% to about 100%, about 20% to about 150%, about 20% to about 200%, about 20% to about 250%, about 50% to about 100%, about 50% to about 150%, about 50% to about 200%, about 50% to about 250%, about 100% to about 150%, about 100% to about 200%, about 100% to about 250%, about 150% to about 200%, about 150% to about 250%, about 200% to about 250%, at least about 10%, at least about 20%, at least about 50%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, or at least about 300%, compared to the amount of mature RNA produced in an untreated cell, e.g., an untreated cell or a cell treated with a control compound. In some embodiments, the total amount of the mRNA encoding OPA1 protein, or the mature mRNA encoding OPA1 protein produced in the cell to which the antisense oligomer is contacted is increased about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9- fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9- fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1- fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold compared to the amount of mature RNA produced in an untreated cell, e.g., an untreated cell or a cell treated with a control compound. A control compound can be, for example, an oligonucleotide that is not complementary to a targeted portion of the 0PA1 NMD exon-containing pre-mRNA.

[0249] The NMD exon can be in any length. In some embodiments, the NMD exon comprises a full sequence of an intron, in which case, it can be referred to as intron retention. In some embodiments, the NMD exon can be a portion of the intron. In some embodiments, the NMD exon can be a 5’ end portion of an intron including a 5’ss sequence. In some embodiments, the NMD exon can be a 3’ end portion of an intron including a 3’ss sequence. In some embodiments, the NMD exon can be a portion within an intron without inclusion of a 5’ss sequence. In some embodiments, the NMD exon can be a portion within an intron without inclusion of a 3’ss sequence. In some embodiments, the NMD exon can be a portion within an intron without inclusion of either a 5’ss or a 3’ss sequence. In some embodiments, the NMD exon can be from 5 nucleotides to 10 nucleotides in length, from 10 nucleotides to 15 nucleotides in length, from 15 nucleotides to 20 nucleotides in length, from 20 nucleotides to 25 nucleotides in length, from 25 nucleotides to 30 nucleotides in length, from 30 nucleotides to 35 nucleotides in length, from 35 nucleotides to 40 nucleotides in length, from 40 nucleotides to 45 nucleotides in length, from 45 nucleotides to 50 nucleotides in length, from 50 nucleotides to 55 nucleotides in length, from 55 nucleotides to 60 nucleotides in length, from 60 nucleotides to 65 nucleotides in length, from 65 nucleotides to 70 nucleotides in length, from 70 nucleotides to 75 nucleotides in length, from 75 nucleotides to 80 nucleotides in length, from 80 nucleotides to 85 nucleotides in length, from 85 nucleotides to 90 nucleotides in length, from 90 nucleotides to 95 nucleotides in length, or from95 nucleotides to 100 nucleotides in length. In some embodiments, the NMD exon can be at least 10 nucleotides, at least 20 nucleotides, at least 30 nucleotides, at least 40 nucleotides, at least 50 nucleotides, at least 60 nucleoids, at least 70 nucleotides, at least 80 nucleotides in length, at least 90 nucleotides, or at least 100 nucleotides in length. In some embodiments, the NMD exon can be from 100 to 200 nucleotides in length, from 200 to 300 nucleotides in length, from 300 to 400 nucleotides in length, from 400 to 500 nucleotides in length, from 500 to 600 nucleotides in length, from 600 to 700 nucleotides in length, from 700 to 800 nucleotides in length, from 800 to 900 nucleotides in length, from 900 to 1,000 nucleotides in length. In some embodiments, the NMD exon may be longer than 1,000 nucleotides in length.

[0250] Inclusion of a pseudo-exon can lead to a frameshift and the introduction of a premature termination codon (PTC) in the mature mRNA transcript rendering the transcript a target of NMD. Mature mRNA transcript containing NMD exon can be nonproductive mRNA transcript which does not lead to protein expression. The PTC can be present in any position downstream of an NMD exon. In some embodiments, the PTC can be present in any exon downstream of an NMD exon. In some embodiments, the PTC can be present within the NMD exon. For example, inclusion of exon 6x of OPA1, exon 7x of OPA1, or exon 28x of OPA1, in an mRNA transcript encoded by the OPA1 gene can induce a PTC in the mRNA transcript. For example, inclusion of exon (GRCh38 / hg38: chr3 193628509 193628616) of OPAL, or exon (GRCh38 / hg38: chr3 193603500 193603557) of OPA1 in an mRNA transcript encoded by the OPAL

[0251] In some aspects, provided herein is a method of modulating expression of an OPA1 protein by promoting inclusion of a coding exon. The method can comprise contacting an agent to a cell having an OPA1 pre-mRNA, wherein the agent comprises an oligonucleotide that binds to: (a) a targeted portion of the pre-mRNA within an intronic region immediately upstream of a 5’ end of the coding exon of the pre-mRNA; or (b) a targeted portion of the pre-mRNA within an intronic region immediately downstream of a 3 ’ end of the coding exon of the pre-mRNA; whereby the agent increases a level of a processed mRNA that is processed from the pre-mRNA and that contains the coding exon in the cell. In some cases, the coding exon to be included is an alternatively spliced exon. In some cases, the method promotes inclusion of the coding exon in the processed mRNA during splicing of the pre-mRNA in the cell.

[0252] In some of these embodiments for inclusion of coding exon, the target portion of the pre-mRNA is within a region spanning from 100 to 50, from 100 to 60, from 100 to 70, from 100 to 80, or from 100 to 90 nucleotides upstream of a 5’ end of the coding exon. In some cases, the target portion of the pre-mRNA is within a region spanning from 40 to 100, from 50 to 100, from 60 to 100, from 70 to 100, from 80 to 100, or from 90 to 100 nucleotides downstream of a 3’ end of the coding exon. In some cases, the coding exon is exon 7 of OPAL In some cases, the coding exon comprises a sequence with at least 80%, at least 90%, or 100% sequence identity to SEQ ID NO: 277. In some cases, the coding exon comprises SEQ ID NO: 277. The targeted portion of the pre-mRNA can be within a region spanning from 100 to 50, from 100 to 60, from 100 to 70, from 100 to 80, or from 100 to 90 nucleotides upstream of genomic site GROG 8 / hg38: chr3 193626092. In some cases, the targeted portion of the pre-mRNA is within a region spanning from 40 to 100, from 50 to 100, from 60 to 100, from 70 to 100, from 80 to 100, or from 90 to 100 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193626202.

[0253] In some cases, the inclusion of the coding exon in the processed mRNA in the cell contacted with the agent is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5- fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5- fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of the agent.Exclusion of Both NMD Exon and Coding Exon

[0254] In some embodiments, provided herein is a method of modulating expression of a target protein by targeting a pre-mRNA and modulating exclusion of both a coding exon and a nonsense-mediated RNA decay-inducing exon (NMD exon) from the pre-mRNA. In some cases, the method comprises contacting an agent to the cell, and the agent promotes exclusion of both the coding exon and the NMD exon from the pre-mRNA, thereby increasing level of a processed mRNA that is processed from the pre-mRNA and lacks both the coding exon and the NMD exon. In some cases, the agent binds to a targeted portion of the pre-mRNA, or modulates binding of a factor involved in splicing of the coding exon, the NMD exon, or both. In some cases, the agent interferes with binding of the factor involved in splicing of the coding exon, the NMD exon, or both, to a region of the targeted portion. In some cases, the NMD exon is within an intronic region adjacent to the coding exon. In some cases, the NMD exon is within an intronic region immediately upstream of the coding exon. In some cases, the NMD exon is within an intronic region immediately downstream of the coding exon. In some cases, the coding exon is an alternatively spliced exon.

[0255] In some cases, the targeted portion of the pre-mRNA is proximal to the coding exon. The targeted portion of the pre-mRNA can be located in an intronic region immediately upstream of the coding exon. The targeted portion of the pre-mRNA can be located in an intronic region immediately downstream of the coding exon. In some cases, the targeted portion of the pre- mRNA can be located within the coding exon. In some cases, the targeted portion of the pre- mRNA is within a region spanning from 49 to 1, from 39 to 1, from 29 to 1, or from 19 to 1 nucleotides upstream of 5’ end of the coding exon. In some cases, the targeted portion of the pre- mRNA is within a region spanning from 100 nucleotides upstream of the coding exon to 100 nucleotides downstream of the coding exon. In some cases, the targeted portion comprises about1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more consecutive nucleotides of the coding exon.

[0256] In some cases, the targeted portion of the pre-mRNA is proximal to the NMD exon. In some cases, the targeted portion of the pre-mRNA is located in an intronic region immediately upstream of the NMD exon. In some cases, the targeted portion of the pre-mRNA is located in an intronic region immediately downstream of the NMD exon. In some cases, the targeted portion of the pre-mRNA is located within the NMD exon. In some cases, the targeted portion of the pre- mRNA is within a region spanning from 100 nucleotides upstream of the NMD exon to 100 nucleotides downstream of the NMD exon.

[0257] In some embodiments, the method described herein is applicable to modulation of expression of OPA1 protein by modulating exclusion of both exon 7 and an NMD exon (e.g., exon 7x) of OPA1 pre-mRNA that contains both exon 7 and exon 7x. In some cases, the coding exon comprises a sequence with at least 80%, at least 90%, or 100% sequence identity to SEQ ID NO: 277. In some cases, the coding exon comprises SEQ ID NO: 277. In some cases, the targeted portion of the pre-mRNA is immediately upstream of the coding exon GRCh38 / hg38: chr3 193626092 to 193626202. In some cases, the targeted portion of the pre-mRNA is immediately downstream of the coding exon GRCh38 / hg38: chr3 193626092 to 193626202. In some cases, the targeted portion of the pre-mRNA is within a region spanning from 49 to 1, from 39 to 1, from 29 to 1, or from 19 to 1 nucleotides upstream of GRCh38 / hg38: chr3 193626092. In some cases, the targeted portion of the pre-mRNA is within a region spanning from 100 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193626092 to 100 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193626202. In some cases, the targeted portion of the pre-mRNA is within the coding exon GRCh38 / hg38: chr3 193626092 to 193626202. In some cases, the targeted portion of the pre-mRNA comprises an exon-intron junction of the coding exon GRCh38 / hg38: chr3 193626092 to 193626202. In some cases, the NMD exon comprises a sequence with at least 80%, at least 90%, or 100% sequence identity to SEQ ID NO: 279. In some cases, the NMD exon comprises SEQ ID NO: 279. In some cases, the targeted portion of the pre-mRNA is immediately upstream of the NMD exon GRCh38 / hg38: chr3 193628509 to 193628616. In some cases, the targeted portion of the pre-mRNA is immediately downstream of the NMD exon GRCh38 / hg38: chr3 193628509 to 193628616. In some cases, the targeted portion of the pre-mRNA is within a region spanning from 100 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193628509 to 100 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193628616.

[0258] In some cases, the targeted portion of the pre-mRNA is within the NMD exon GRCh38 / hg38: chr3 193628509 to 193628616. In some cases, the targeted portion of the pre- mRNA comprises an exon-intron junction of the NMD exon GRCh38 / hg38: chr3 193628509 to193628616. In some cases, the targeted portion comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more consecutive nucleotides of the NMD exon.

[0259] In some cases, the exclusion of the coding exon from the pre-mRNA in the cell contacted with the agent is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5- fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5- fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of contacting with the agent. In some cases, the exclusion of the NMD exon from the pre-mRNA in the cell contacted with the agent is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1 -fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5- fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of contacting with the agent. In some cases, the method results in an increase in the level of the processed mRNA in the cell. The level of the processed mRNA in the cell contacted with the agent can be increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10- fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of contacting with the agent.

[0260] In some cases, the method results in an increase in expression of the OPA1 protein in the cell. A level of the OPA1 protein expressed from the processed mRNA in the cell contactedwith the agent can be increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of contacting with the agent.

[0261] In some cases, a level of the OPA1 protein expressed from the processed mRNA in the cell contacted with the agent is increased by at least about 1.5-fold compared to in the absence of contacting with the agent.

[0262] In some cases, the OP Al protein expressed from the processed mRNA that lacks exon 7 and exon 7x is a functional OPA1 protein. The OPA1 protein expressed from the processed mRNA that lacks exon 7 and exon 7x can be at least partially functional as compared to a wildtype OPA1 protein. The OPA1 protein expressed from the processed mRNA that lacks exon 7 and exon 7x can be at least partially functional as compared to a full-length wild-type OPA1 protein.Therapeutic Agents

[0263] In various embodiments of the present disclosure, compositions and methods comprising a therapeutic agent are provided to modulate protein expression level of OPA1. In some embodiments, provided herein are compositions and methods to modulate alternative splicing of OPA1 pre-mRNA. In some embodiments, provided herein are compositions and methods to induce exon skipping in the splicing of OPA1 pre-mRNA, e.g., to induce skipping of a pseudo-exon during splicing of OPA1 pre-mRNA. In other embodiments, therapeutic agents may be used to induce the inclusion of an exon in order to decrease the protein expression level.

[0264] A therapeutic agent disclosed herein can be a NIE repressor agent. A therapeutic agent may comprise a polynucleic acid polymer.

[0265] According to one aspect of the present disclosure, provided herein is a method of treatment or prevention of a condition or disease associated with a functional OPA1 protein deficiency, comprising administering a NIE repressor agent to a subject to increase levels of functional OPA1 protein, wherein the agent binds to a region of the pre-mRNA transcript to decrease inclusion of the NMD exon in the mature transcript. For example, provided herein is a method of treatment or prevention of a condition associated with a functional OPA1 protein deficiency, comprising administering a NIE repressor agent to a subject to increase levels of functional OPA1 protein, wherein the agent binds to a region of an intron containing an NMDexon e.g., exon 6x of OPA1, exon 7x of OPA1, or exon 28x of OPAP) of the pre-mRNA transcript or to a NMD exon-activating regulatory sequence in the same intron. For example, provided herein is a method of treatment or prevention of a condition associated with a functional 0PA1 protein deficiency, comprising administering a NIE repressor agent to a subject to increase levels of functional 0PA1 protein, wherein the agent binds to a region of an intron containing an NMD exon (e.g., exon (GRCh38 / hg38: chr3 193628509 193628616) of OPAP, or exon (GRCh38 / hg38: chr3 193603500 193603557) of OP Al) of the pre-mRNA transcript or to a NMD exon-activating regulatory sequence in the same intron. In some embodiments, the method comprises administering a NIE repressor agent to a subject to increase levels of functional OPA1 protein, wherein the agent binds to a region of an intron containing an NMD exon (e.g., exon of OPA1 other than exon 7x defined by (GRCh38 / hg38: chr3 193628509 193628616) or exon defined by (GROG 8 / hg38: chr3 193603500 193603557)) of the pre-mRNA transcript or to a NMD exonactivating regulatory sequence in the same intron. In some embodiments, the therapeutic agent promotes exclusion of an NMD exon of OPA1 pre-mRNA other than exon 7x defined by (GRCh38 / hg38: chr3 193628509 193628616) or exon defined by (GRO138 / hg38: chr3 193603500 193603557). In some embodiments, the composition disclosed herein includes an agent that promotes exclusion of an NMD exon of OPA1 pre-mRNA other than exon 7x defined by (GRCh38 / hg38: chr3 193628509 193628616) or exon defined by (GRCh38 / hg38: chr3 193603500 193603557).

[0266] Where reference is made to reducing NMD exon inclusion in the mature mRNA, the reduction may be complete, e.g., 100%, or may be partial. The reduction may be clinically significant. The reduction / correction may be relative to the level of NMD exon inclusion in the subject without treatment, or relative to the amount of NMD exon inclusion in a population of similar subjects. The reduction / correction may be at least 10% less NMD exon inclusion relative to the average subject, or the subject prior to treatment. The reduction may be at least 20% less NMD exon inclusion relative to an average subject, or the subject prior to treatment. The reduction may be at least 40% less NMD exon inclusion relative to an average subject, or the subject prior to treatment. The reduction may be at least 50% less NMD exon inclusion relative to an average subject, or the subject prior to treatment. The reduction may be at least 60% less NMD exon inclusion relative to an average subject, or the subject prior to treatment. The reduction may be at least 80% less NMD exon inclusion relative to an average subject, or the subject prior to treatment. The reduction may be at least 90% less NMD exon inclusion relative to an average subject, or the subject prior to treatment.

[0267] According to one aspect of the present disclosure, provided herein is a method of treatment or prevention of a condition or disease associated with a functional OPA1 proteindeficiency, comprising administering an agent to a subject to increase levels of functional 0PA1 protein, wherein the agent binds to a region of the pre-mRNA transcript to decrease inclusion of a coding exon (e.g., exon 7) in the mature transcript. For example, provided herein is a method of treatment or prevention of a condition associated with a functional 0PA1 protein deficiency, comprising administering an agent to a subject to increase levels of functional 0PA1 protein, wherein the agent binds to a region containing a coding exon (e.g., exon 7 of OPAP) of the pre- mRNA transcript. For example, provided herein is a method of treatment or prevention of a condition associated with a functional 0PA1 protein deficiency, comprising administering an agent to a subject to increase levels of functional 0PA1 protein, wherein the agent binds to a region containing a coding exon (e.g., exon (GRCh38 / hg38: chr3 193626092 to 193626202) of OPA L) of the pre-mRNA transcript. In some embodiments, the method comprises administering an agent to a subject to increase levels of functional OPA1 protein, wherein the agent binds to a region containing a coding exon (e.g., exon of OPA1 other than exon 7 defined by (GRCh38 / hg38: chr3 193626092 to 193626202)) of the pre-mRNA transcript. In some embodiments, the therapeutic agent promotes exclusion of a coding exon of OPA1 pre-mRNA other than exon 7 defined by (GROG 8 / hg38: chr3 193626092 to 193626202). In some embodiments, the composition disclosed herein includes an agent that promotes exclusion of a coding exon of OPA1 pre-mRNA other than exon 7 defined by (GROG 8 / hg38: chr3 193626092 to 193626202).

[0268] Where reference is made to increasing active OPA1 protein levels, the increase may be clinically significant. The increase may be relative to the level of active OPA1 protein in the subject without treatment, or relative to the amount of active OPA1 protein in a population of similar subjects. The increase may be at least 10% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 20% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 40% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 50% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 80% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 100% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 200% more active OPA1 protein relative to the average subject, or the subject prior to treatment. The increase may be at least 500% more active OPA1 protein relative to the average subject, or the subject prior to treatment.

[0269] In embodiments wherein the NIE repressor agent comprises a polynucleic acid polymer, the polynucleic acid polymer may be about 50 nucleotides in length. The polynucleic acid polymer may be about 45 nucleotides in length. The polynucleic acid polymer may be about40 nucleotides in length. The polynucleic acid polymer may be about 35 nucleotides in length. The polynucleic acid polymer may be about 30 nucleotides in length. The polynucleic acid polymer may be about 24 nucleotides in length. The polynucleic acid polymer may be about 25 nucleotides in length. The polynucleic acid polymer may be about 20 nucleotides in length. The polynucleic acid polymer may be about 19 nucleotides in length. The polynucleic acid polymer may be about18 nucleotides in length. The polynucleic acid polymer may be about 17 nucleotides in length. The polynucleic acid polymer may be about 16 nucleotides in length. The polynucleic acid polymer may be about 15 nucleotides in length. The polynucleic acid polymer may be about 14 nucleotides in length. The polynucleic acid polymer may be about 13 nucleotides in length. The polynucleic acid polymer may be about 12 nucleotides in length. The polynucleic acid polymer may be about11 nucleotides in length. The polynucleic acid polymer may be about 10 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 50 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 45 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 40 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 35 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 30 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 25 nucleotides in length. The polynucleic acid polymer may be between about 10 and about 20 nucleotides in length. The polynucleic acid polymer may be between about 15 and about 25 nucleotides in length. The polynucleic acid polymer may be between about 15 and about 30 nucleotides in length. The polynucleic acid polymer may be between about 12 and about 30 nucleotides in length.

[0270] The sequence of the polynucleic acid polymer may be at least 50%, 55%, 60%,65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% complementary to a target sequence of an mRNA transcript, e.g., a partially processed mRNA transcript. The sequence of the polynucleic acid polymer may be 100% complementary to a target sequence of a pre-mRNA transcript.

[0271] The sequence of the polynucleic acid polymer may have 4 or fewer mismatches to a target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 3 or fewer mismatches to a target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 2 or fewer mismatches to a target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have 1 or fewer mismatches to a target sequence of the pre-mRNA transcript. The sequence of the polynucleic acid polymer may have no mismatches to a target sequence of the pre-mRNA transcript.

[0272] The polynucleic acid polymer may specifically hybridize to a target sequence of the pre-mRNA transcript. For example, the polynucleic acid polymer may have 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 100% sequence complementarity to a target sequence of the pre-mRNA transcript. The hybridization may be under high stringent hybridization conditions.

[0273] The polynucleic acid polymer comprising a sequence with at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.5% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 2-5. The polynucleic acid polymer may comprise a sequence with 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 2-5.

[0274] Where reference is made to a polynucleic acid polymer sequence, the skilled person will understand that one or more substitutions may be tolerated, optionally two substitutions may be tolerated in the sequence, such that it maintains the ability to hybridize to the target sequence; or where the substitution is in a target sequence, the ability to be recognized as the target sequence. References to sequence identity may be determined by BLAST sequence alignment using standard / default parameters. For example, the sequence may have 99% identity and still function according to the present disclosure. In other embodiments, the sequence may have 98% identity and still function according to the present disclosure. In another embodiment, the sequence may have 95% identity and still function according to the present disclosure. In another embodiment, the sequence may have 90% identity and still function according to the present disclosure.Antisense Oligomers

[0275] Provided herein is a composition comprising an antisense oligomer that induces exon skipping by binding to a targeted portion of an OPA1 pre-mRNA, e.g., an OPA1 NMD exoncontaining pre-mRNA. As used herein, the terms “ASO” and “antisense oligomer” are used interchangeably and refer to an oligomer such as a polynucleotide, comprising nucleobases that hybridizes to a target nucleic acid (e.g., an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA) sequence by Watson-Crick base pairing or wobble base pairing (G-U). The ASO may have exact sequence complementary to the target sequence or near complementarity (e.g., sufficient complementarity to bind the target sequence and enhancing splicing at a splice site). ASOs are designed so that they bind (hybridize) to a target nucleic acid (e.g., a targeted portion of a pre-mRNA transcript) and remain hybridized under physiological conditions. Typically, if they hybridize to a site other than the intended (targeted) nucleic acid sequence, they hybridize to a limited number of sequences that are not a target nucleic acid (to a few sites other than a target nucleic acid). Design of an ASO can take into consideration the occurrence of the nucleic acid sequence of the targeted portion of the pre-mRNA transcript or a sufficiently similar nucleic acid sequence in other locations in the genome or cellular pre-mRNA or transcriptome, such that the likelihood the ASO will bind other sites and cause “off-target” effects is limited. Any antisenseoligomers known in the art (for example, in PCT Application No. PCT / US2014 / 054151, published as WO 2015 / 035091, titled “Reducing Nonsense-Mediated mRNA Decay,” incorporated by reference herein), can be used to practice the methods described herein.

[0276] In some embodiments, ASOs “specifically hybridize” to or are “specific” to a target nucleic acid or a targeted portion of an OPA1 pre-mRNA, e.g., a NMD exon-containing pre- mRNA. Typically, such hybridization occurs with a Tmsubstantially greater than 37 °C, preferably at least 50 °C, and typically between 60 °C to approximately 90 °C. Such hybridization preferably corresponds to stringent hybridization conditions. At a given ionic strength and pH, the Tmis the temperature at which 50% of a target sequence hybridizes to a complementary oligonucleotide.

[0277] Oligomers, such as oligonucleotides, are “complementary” to one another when hybridization occurs in an antiparallel configuration between two single-stranded polynucleotides. A double-stranded polynucleotide can be “complementary” to another polynucleotide, if hybridization can occur between one of the strands of the first polynucleotide and the second. Complementarity (the degree to which one polynucleotide is complementary with another) is quantifiable in terms of the proportion (e.g., the percentage) of bases in opposing strands that are expected to form hydrogen bonds with each other, according to generally accepted base-pairing rules. The sequence of an antisense oligomer (ASO) need not be 100% complementary to that of its target nucleic acid to hybridize. In certain embodiments, ASOs can comprise at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence complementarity to a target region within the target nucleic acid sequence to which they are targeted. For example, an ASO in which 18 of 20 nucleobases of the oligomeric compound are complementary to a target region, and would therefore specifically hybridize, would represent 90 percent complementarity. In this example, the remaining non- complementary nucleobases may be clustered together or interspersed with complementary nucleobases and need not be contiguous to each other or to complementary nucleobases. Percent complementarity of an ASO with a region of a target nucleic acid can be determined routinely using BLAST programs (basic local alignment search tools) and PowerBLAST programs known in the art (Altschul, et al., J. Mol. Biol., 1990, 215, 403-410; Zhang and Madden, Genome Res., 1997, 7, 649-656).

[0278] An ASO need not hybridize to all nucleobases in a target sequence and the nucleobases to which it does hybridize may be contiguous or noncontiguous. ASOs may hybridize over one or more segments of a pre-mRNA transcript, such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure or hairpin structure may be formed). In certain embodiments, an ASO hybridizes to noncontiguous nucleobases in a target pre-mRNA transcript. For example, an ASO can hybridize to nucleobases in a pre-mRNA transcript that are separated by one or more nucleobase(s) to which the ASO does not hybridize.

[0279] The ASOs described herein comprise nucleobases that are complementary to nucleobases present in a target portion of an OPA1 pre-mRNA, e.g., a NMD exon-containing pre- mRNA. The term ASO embodies oligonucleotides and any other oligomeric molecule that comprises nucleobases capable of hybridizing to a complementary nucleobase on a target mRNA but does not comprise a sugar moiety, such as a peptide nucleic acid (PNA). The ASOs may comprise naturally-occurring nucleotides, nucleotide analogs, modified nucleotides, or any combination of two or three of the preceding. The term “naturally occurring nucleotides” includes deoxyribonucleotides and ribonucleotides. The term “modified nucleotides” includes nucleotides with modified or substituted sugar groups and / or having a modified backbone. In some embodiments, all of the nucleotides of the ASO are modified nucleotides. Chemical modifications of ASOs or components of ASOs that are compatible with the methods and compositions described herein will be evident to one of skill in the art and can be found, for example, in U.S. Patent No. 8,258,109 B2, U.S. Patent No. 5,656,612, U.S. Patent Publication No. 2012 / 0190728, and Dias and Stein, Mol. Cancer Ther. 2002, 347-355, herein incorporated by reference in their entirety.

[0280] One or more nucleobases of an ASO may be any naturally occurring, unmodified nucleobase such as adenine, guanine, cytosine, thymine and uracil, or any synthetic or modified nucleobase that is sufficiently similar to an unmodified nucleobase such that it is capable of hydrogen bonding with a nucleobase present on a target pre-mRNA. Examples of modified nucleobases include, without limitation, hypoxanthine, xanthine, 7-methylguanine, 5, 6- dihydrouracil, 5-methylcytosine, and 5-hydroxymethoylcytosine.

[0281] The ASOs described herein also comprise a backbone structure that connects the components of an oligomer. The term “backbone structure” and “oligomer linkages” may be used interchangeably and refer to the connection between monomers of the ASO. In naturally occurring oligonucleotides, the backbone comprises a 3 ’-5’ phosphodiester linkage connecting sugar moieties of the oligomer. The backbone structure or oligomer linkages of the ASOs described herein may include, but are not limited to, phosphorothioate, phosphorodithioate, phosphoroselenoate, phosphorodiselenoate, phosphoroanilothioate, phosphoraniladate, phosphoramidate, and the like. See, e.g., LaPlanche, et al., Nucleic Acids Res. 14:9081 (1986); Stec, et al., J. Am. Chem. Soc. 106:6077 (1984), Stein, et al., Nucleic Acids Res. 16:3209 (1988), Zon, et al., Anti-Cancer Drug Design 6:539 (1991); Zon, et al., Oligonucleotides and Analogues: A Practical Approach, pp. 87-108 (F. Eckstein, Ed., Oxford University Press, Oxford England (1991)); Stec, et al., U.S. Pat. No. 5,151,510; Uhlmann and Peyman, Chemical Reviews 90:543 (1990). In some embodiments, the backbone structure of the ASO does not contain phosphorousbut rather contains peptide bonds, for example, in a peptide nucleic acid (PNA), or linking groups including carbamate, amides, and linear and cyclic hydrocarbon groups. In some embodiments, the backbone modification is a phosphorothioate linkage. In some embodiments, the backbone modification is a phosphoramidate linkage.

[0282] In some embodiments, the stereochemistry at each of the phosphorus internucleotide linkages of the ASO backbone is random. In some embodiments, the stereochemistry at each of the phosphorus intemucleotide linkages of the ASO backbone is controlled and is not random. For example, U.S. Pat. App. Pub. No. 2014 / 0194610, “Methods for the Synthesis of Functionalized Nucleic Acids,” incorporated herein by reference, describes methods for independently selecting the handedness of chirality at each phosphorous atom in a nucleic acid oligomer. In some embodiments, an ASO used in the methods of the disclosure, including, but not limited to, any of the ASOs set forth herein in Tables 5 and 6, comprises an ASO having phosphorus internucleotide linkages that are not random. In some embodiments, a composition used in the methods of the disclosure comprises a pure diastereomeric ASO. In some embodiments, a composition used in the methods of the disclosure comprises an ASO that has diastereomeric purity of at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, about 100%, about 90% to about 100%, about 91% to about 100%, about 92% to about 100%, about 93% to about 100%, about 94% to about 100%, about 95% to about 100%, about 96% to about 100%, about 97% to about 100%, about 98% to about 100%, or about 99% to about 100%.

[0283] In some embodiments, the ASO has a nonrandom mixture of Rp and Sp configurations at its phosphorus internucleotide linkages. For example, it has been suggested that a mix of Rp and Sp is required in antisense oligonucleotides to achieve a balance between good activity and nuclease stability (Wan, etal., 2014, “Synthesis, biophysical properties and biological activity of second-generation antisense oligonucleotides containing chiral phosphorothioate linkages,” Nucleic Acids Research 42(22): 13456-13468, incorporated herein by reference). In some embodiments, an ASO used in the methods of the disclosure, including, but not limited to, any of the ASOs set forth herein in SEQ ID NOs: 2-5, comprises about 5-100% Rp, at least about 5% Rp, at least about 10% Rp, at least about 15% Rp, at least about 20% Rp, at least about 25% Rp, at least about 30% Rp, at least about 35% Rp, at least about 40% Rp, at least about 45% Rp, at least about 50% Rp, at least about 55% Rp, at least about 60% Rp, at least about 65% Rp, at least about 70% Rp, at least about 75% Rp, at least about 80% Rp, at least about 85% Rp, at least about 90% Rp, or at least about 95% Rp, with the remainder Sp, or about 100% Rp. In some embodiments, an ASO used in the methods of the disclosure, including, but not limited to, any ofthe ASOs set forth herein comprise a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5, comprises about 10% to about 100% Rp, about 15% to about 100% Rp, about 20% to about 100% Rp, about 25% to about 100% Rp, about 30% to about 100% Rp, about 35% to about 100% Rp, about 40% to about 100% Rp, about 45% to about 100% Rp, about 50% to about 100% Rp, about 55% to about 100% Rp, about 60% to about 100% Rp, about 65% to about 100% Rp, about 70% to about 100% Rp, about 75% to about 100% Rp, about 80% to about 100% Rp, about 85% to about 100% Rp, about 90% to about 100% Rp, or about 95% to about 100% Rp, about 20% to about 80% Rp, about 25% to about 75% Rp, about 30% to about 70% Rp, about 40% to about 60% Rp, or about 45% to about 55% Rp, with the remainder Sp.

[0284] In some embodiments, an ASO used in the methods of the disclosure, including, but not limited to, any of the ASOs set forth herein comprise a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5, comprises about 5-100% Sp, at least about 5% Sp, at least about 10% Sp, at least about 15% Sp, at least about 20% Sp, at least about 25% Sp, at least about 30% Sp, at least about 35% Sp, at least about 40% Sp, at least about 45% Sp, at least about 50% Sp, at least about 55% Sp, at least about 60% Sp, at least about 65% Sp, at least about 70% Sp, at least about 75% Sp, at least about 80% Sp, at least about 85% Sp, at least about 90% Sp, or at least about 95% Sp, with the remainder Rp, or about 100% Sp. In embodiments, an ASO used in the methods of the disclosure, including, but not limited to, any of the ASOs set forth herein comprise a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of any one of SEQ ID NOs: 2-5, comprises about 10% to about 100% Sp, about 15% to about 100% Sp, about 20% to about 100% Sp, about 25% to about 100% Sp, about 30% to about 100% Sp, about 35% to about 100% Sp, about 40% to about 100% Sp, about 45% to about 100% Sp, about 50% to about 100% Sp, about 55% to about 100% Sp, about 60% to about 100% Sp, about 65% to about 100% Sp, about 70% to about 100% Sp, about 75% to about 100% Sp, about 80% to about 100% Sp, about 85% to about 100% Sp, about 90% to about 100% Sp, or about 95% to about 100% Sp, about 20% to about 80% Sp, about 25% to about 75% Sp, about 30% to about 70% Sp, about 40% to about 60% Sp, or about 45% to about 55% Sp, with the remainder Rp.

[0285] Any of the ASOs described herein may contain a sugar moiety that comprises ribose or deoxyribose, as present in naturally occurring nucleotides, or a modified sugar moiety or sugar analog, including a morpholine ring. Non-limiting examples of modified sugar moieties include 2’ substitutions such as 2’-O-methyl (2’-0-Me), 2’ -O-m ethoxy ethyl (2’MOE), 2’-O- aminoethyl, 2’F; N3’->P5’ phosphoramidate, 2’dimethylaminooxyethoxy,2’dimethylaminoethoxyethoxy, 2’-guanidinidium, 2’-0-guanidinium ethyl, carbamate modified sugars, and bicyclic modified sugars. In some embodiments, the sugar moiety modification is selected from 2’-0-Me, 2’F, and 2’MOE. In some embodiments, the sugar moiety modification is an extra bridge bond, such as in a locked nucleic acid (LNA). In some embodiments the sugar analog contains a morpholine ring, such as phosphorodiamidate morpholino (PMO). In some embodiments, the sugar moiety comprises a ribofuranosyl or 2’deoxyribofuransyl modification. In some embodiments, the sugar moiety comprises 2’4’ -constrained 2’0-methyloxyethyl (cMOE) modifications. In some embodiments, the sugar moiety comprises cEt 2’, 4’ constrained 2’-0 ethyl BNA modifications. In some embodiments, the sugar moiety comprises tricycloDNA (tcDNA) modifications. In some embodiments, the sugar moiety comprises ethylene nucleic acid (ENA) modifications. In some embodiments, the sugar moiety comprises MCE modifications. Modifications are known in the art and described in the literature, e.g., by Jarver, et al., 2014, “A Chemical View of Oligonucleotides for Exon Skipping and Related Drug Applications,” Nucleic Acid Therapeutics 24(1): 37-47, incorporated by reference for this purpose herein.

[0286] In some embodiments, each monomer of the ASO is modified in the same way, for example each linkage of the backbone of the ASO comprises a phosphorothioate linkage or each ribose sugar moiety comprises a 2’0-methyl modification. Such modifications that are present on each of the monomer components of an ASO are referred to as “uniform modifications.” In some examples, a combination of different modifications may be desired, for example, an ASO may comprise a combination of phosphorodiamidate linkages and sugar moieties comprising morpholine rings (morpholinos). Combinations of different modifications to an ASO are referred to as “mixed modifications” or “mixed chemistries.”

[0287] In some embodiments, the ASO comprises one or more backbone modifications. In some embodiments, the ASO comprises one or more sugar moiety modifications. In some embodiments, the ASO comprises one or more backbone modifications and one or more sugar moiety modifications. In some embodiments, the ASO comprises a 2’MOE modification and a phosphorothioate backbone. In some embodiments, the ASO comprises a phosphorodiamidate morpholino (PMO). In some embodiments, the ASO comprises a peptide nucleic acid (PNA). Any of the ASOs or any component of an ASO (e.g., a nucleobase, sugar moiety, backbone) described herein may be modified in order to achieve desired properties or activities of the ASO or reduce undesired properties or activities of the ASO. For example, an ASO or one or more components of any ASO may be modified to enhance binding affinity to a target sequence on a pre-mRNA transcript; reduce binding to any non-target sequence; reduce degradation by cellular nucleases (i.e., RNase H); improve uptake of the ASO into a cell and / or into the nucleus of a cell; alter the pharmacokinetics or pharmacodynamics of the ASO; and / or modulate the half-life of the ASO.

[0288] In some embodiments, the ASOs are comprised of 2'-O-(2 -methoxyethyl) (MOE) phosphorothioate-modified nucleotides. ASOs comprised of such nucleotides are especially well- suited to the methods disclosed herein; oligomers having such modifications have been shown to have significantly enhanced resistance to nuclease degradation and increased bioavailability, making them suitable, for example, for oral delivery in some embodiments described herein. See e.g., Geary, etal., J Pharmacol Exp Ther. 2001; 296(3):890-7; Geary, etal., JPharmacol Exp Ther. 2001; 296(3):898-904.

[0289] In some embodiments, the ASOs comprise 5 ’-methylcytosine (5’-MeC) nucleotides. In some embodiments, the ASOs comprise of at least one 5 ’-methylcytosine (5’-MeC) nucleotide. In some embodiments, each cytosine of the ASO is a 5 ’-methylcytosine (5’-MeC). In some embodiments, the ASOs comprise a 5 ’-methyluracil (5’-MeU). In some embodiments, the ASOs comprise at least one 5 ’-methyluracil (5’-MeU). In some embodiments, at least one cytosine or thymidine of the antisense oligomer is a 5 ’-methyluracil (5’-MeU). In some embodiments, each cytosine or thymidine of the antisense oligomer is a 5 ’-methyluracil (5’-MeU).

[0290] In any of the structural formulae (graphic representation of a chemical compound) presented herein, where two curved lines and a straight line in between them are used to connect a phosphorus atom (“P”) and an oxygen atom (“O”), the two curved line and the straight line therebetween should be seen as a single integral segment, representing the covalent bond between the phosphorus atom and the oxygen atom, which is part of a backbone linkage (e.g. , phosphodiester linkage or phosphorothioate linkage) between two neighboring nucleotides. Any of the vertices (corners) in any of those structural formulae where a curved line and a straight-line join does not represent carbon atom or presence of -CH2- at the relevant location in the compound the structural formula represents.

[0291] In some embodiments, the ASO has any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0292] In some embodiments, the ASO has the following structure:or a pharmaceutically acceptable salt thereof.

[0293] In some embodiments, the ASO has the following structure:or a pharmaceutically acceptable salt thereof.

[0294] In some embodiments, the ASO has the following structure:or a pharmaceutically acceptable salt thereof.

[0295] In some embodiments, the ASO has the following structure:or a pharmaceutically acceptable salt thereof.

[0296] In some embodiments, the ASO has the following structure:or a pharmaceutically acceptable salt thereof.

[0297] In some embodiments, the ASO has any one of the following structures:

[0298] In some embodiments, the ASO is in a salt form. In some embodiments, the salt form is sodium salt. In some embodiments, the sodium salt form of the ASO has the following structure:

[0299] In some embodiments, the ASO is in a salt form. In some embodiments, the salt form is sodium salt. In some embodiments, the sodium salt form of the ASO has the following structure:

[0300] In some embodiments, the ASO is in a salt form. In some embodiments, the salt form is sodium salt. In some embodiments, the sodium salt form of the ASO has the following structure:

[0301] In some embodiments, the ASO is in a salt form. In some embodiments, the salt form is sodium salt. In some embodiments, the sodium salt form of the ASO has the following structure:

[0302] In some embodiments, the ASO is in a salt form. In some embodiments, the salt form is sodium salt. In some embodiments, the sodium salt form of the ASO has the following structure:

[0303] Methods of synthesizing ASOs will be known to one of skill in the art. Alternatively or in addition, ASOs may be obtained from a commercial source.

[0304] Unless specified otherwise, the left-hand end of single-stranded nucleic acid (e.g., pre-mRNA transcript, oligonucleotide, ASO, etc.) sequences is the 5’ end and the left-hand direction of single or double-stranded nucleic acid sequences is referred to as the 5’ direction. Similarly, the right-hand end or direction of a nucleic acid sequence (single or double stranded) is the 3’ end or direction. Generally, a region or sequence that is 5’ to a reference point in a nucleic acid is referred to as “upstream,” and a region or sequence that is 3 ’ to a reference point in a nucleic acid is referred to as “downstream.” Generally, the 5’ direction or end of an mRNA is where the initiation or start codon is located, while the 3’ end or direction is where the termination codon is located. In some aspects, nucleotides that are upstream of a reference point in a nucleic acid may be designated by a negative number, while nucleotides that are downstream of a reference point may be designated by a positive number. For example, a reference point (e.g., an exon-exon junction in mRNA) may be designated as the “zero” site, and a nucleotide that is directly adjacent and upstream of the reference point is designated “minus one,” e.g., “-1,” while a nucleotide that is directly adjacent and downstream of the reference point is designated “plus one,” e.g., “+1 ”

[0305] In some embodiments, the ASOs are complementary to (and bind to) a targeted portion of an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is downstream (in the 3’ direction) of the 5’ splice site (or 3’ end of the NMD exon) of the included exon in an OP Al pre-mRNA (e.g., the direction designated by positive numbers relative to the 5’ splice site). In some embodiments, the ASOs are complementary to a targeted portion of the OPA1 pre-mRNA, e.g., the OPA1 NMD exon-containing pre-mRNA that is within the region about +1 to about +500 relative to the 5’ splice site (or 3’ end) of the included exon. In some embodiments, the ASOs may be complementary to a targeted portion of an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is within the region between nucleotides +6 and +40,000 relative to the 5’ splice site (or 3’ end) of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region about +1 to about +40,000, about +1 to about +30,000, about +1 to about +20,000, about +1 to about +15,000, about +1 to about +10,000, about +1 to about +5,000, about +1 to about +4,000, about +1 to about +3,000, about +1 to about +2,000, about +1 to about +1,000, about +1 to about +500, about +1 to about +490, about +1 to about +480, about +1 to about +470, about +1 to about +460, about +1 to about +450, about+1 to about +440, about +1 to about +430, about +1 to about +420, about +1 to about +410, about+1 to about +400, about +1 to about +390, about +1 to about +380, about +1 to about +370, about+1 to about +360, about +1 to about +350, about +1 to about +340, about +1 to about +330, about+1 to about +320, about +1 to about +310, about +1 to about +300, about +1 to about +290, about+1 to about +280, about +1 to about +270, about +1 to about +260, about +1 to about +250, about+1 to about +240, about +1 to about +230, about +1 to about +220, about +1 to about +210, about+1 to about +200, about +1 to about +190, about +1 to about +180, about +1 to about +170, about +1 to about +160, about +1 to about +150, about +1 to about +140, about +1 to about +130, about +1 to about +120, about +1 to about +110, about +1 to about +100, about +1 to about +90, about +1 to about +80, about +1 to about +70, about +1 to about +60, about +1 to about +50, about +1 to about +40, about +1 to about +30, or about +1 to about +20 relative to 5’ splice site (or 3’ end) of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region from about +1 to about +100, from about +100 to about +200, from about +200 to about +300, from about +300 to about +400, or from about +400 to about +500 relative to 5’ splice site (or 3’ end) of the included exon.

[0306] In some embodiments, the ASOs are complementary to (and bind to) a targeted portion of an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is upstream (in the 5’ direction) of the 5’ splice site (or 3’ end) of the included exon in an OP Al pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA (e.g., the direction designated by negative numbers relative to the 5’ splice site). In some embodiments, the ASOs are complementary to a targeted portion of the OPA1 pre-mRNA, e.g., the OPA1 NMD exon-containing pre-mRNA, that is within the region about -4 to about -270 relative to the 5’ splice site (or 3 ’end) of the included exon. In some embodiments, the ASOs may be complementary to a targeted portion of an OP Al pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is within the region between nucleotides -1 and -40,000 relative to the 5’ splice site (or 3’ end) of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region about -1 to about -40,000, about -1 to about -30,000, about -1 to about -20,000, about -1 to about -15,000, about -1 to about -10,000, about -1 to about -5,000, about -1 to about -4,000, about -1 to about - 3,000, about -1 to about -2,000, about -1 to about -1,000, about -1 to about -500, about -1 to about -490, about -1 to about -480, about -1 to about -470, about -1 to about -460, about -1 to about - 450, about -1 to about -440, about -1 to about -430, about -1 to about -420, about -1 to about -410, about -1 to about -400, about -1 to about -390, about -1 to about -380, about -1 to about -370, about -1 to about -360, about -1 to about -350, about -1 to about -340, about -1 to about -330, about -1 to about -320, about -1 to about -310, about -1 to about -300, about -1 to about -290, about -1 to about -280, about -1 to about -270, about -1 to about -260, about -1 to about -250, about -1 to about -240, about -1 to about -230, about -1 to about -220, about -1 to about -210, about -1 to about - 200, about -1 to about -190, about -1 to about -180, about -1 to about -170, about -1 to about -160, about -1 to about -150, about -1 to about -140, about -1 to about -130, about -1 to about -120, about -1 to about -110, about -1 to about -100, about -1 to about -90, about -1 to about -80, about -1 to about -70, about -1 to about -60, about -1 to about -50, about -1 to about -40, about -1 to about - 30, or about -1 to about -20 relative to 5’ splice site (or 3’ end) of the included exon.

[0307] In some embodiments, the ASOs are complementary to a targeted region of an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is upstream (in the 5’ direction) of the 3’ splice site (or 5’ end) of the included exon in an OP Al pre-mRNA (e.g., in the direction designated by negative numbers). In some embodiments, the ASOs are complementary to a targeted portion of the OPA1 pre-mRNA, e.g., the OPA1 NMD exon-containing pre-mRNA, that is within the region about -1 to about -500 relative to the 3’ splice site (or 5’ end) of the included exon. In some embodiments, the ASOs are complementary to a targeted portion of the OP Al pre-mRNA that is within the region -1 to -40,000 relative to the 3’ splice site of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region about -1 to about -40,000, about -1 to about -30,000, -1 to about -20,000, about -1 to about -15,000, about -1 to about -10,000, about -1 to about -5,000, about -1 to about -4,000, about -1 to about - 3,000, about -1 to about -2,000, about -1 to about -1,000, about -1 to about -500, about -1 to about -490, about -1 to about -480, about -1 to about -470, about -1 to about -460, about -1 to about - 450, about -1 to about -440, about -1 to about -430, about -1 to about -420, about -1 to about -410, about -1 to about -400, about -1 to about -390, about -1 to about -380, about -1 to about -370, about -1 to about -360, about -1 to about -350, about -1 to about -340, about -1 to about -330, about -1 to about -320, about -1 to about -310, about -1 to about -300, about -1 to about -290, about -1 to about -280, about -1 to about -270, about -1 to about -260, about -1 to about -250, about -1 to about -240, about -1 to about -230, about -1 to about -220, about -1 to about -210, about -1 to about - 200, about -1 to about -190, about -1 to about -180, about -1 to about -170, about -1 to about -160, about -1 to about -150, about -1 to about -140, about -1 to about -130, about -1 to about -120, about -1 to about -110, about -1 to about -100, about -1 to about -90, about -1 to about -80, about -1 to about -70, about -1 to about -60, about -1 to about -50, about -1 to about -40, about -1 to about - 30, or about -1 to about -20 relative to 3’ splice site of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region from about -1 to about - 100, from about -100 to about -200, from about -200 to about -300, from about -300 to about -400, or from about -400 to about -500 relative to 3’ splice site of the included exon.

[0308] In some embodiments, the ASOs are complementary to a targeted region of an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA, that is downstream (in the 3’ direction) of the 3’ splice site (5’ end) of the included exon in an OPA1 pre-mRNA, e.g., an OPA1 NMD exon-containing pre-mRNA (e.g., in the direction designated by positive numbers). In some embodiments, the ASOs are complementary to a targeted portion of the OPA1 pre-mRNA that is within the region of about +1 to about +40,000 relative to the 3’ splice site of the included exon. In some aspects, the ASOs are complementary to a targeted portion that is within the region about +1 to about +40,000, about +1 to about +30,000, about +1 to about +20,000, about +1 to about+15,000, about +1 to about +10,000, about +1 to about +5,000, about +1 to about +4,000, about+1 to about +3,000, about +1 to about +2,000, about +1 to about +1,000, about +1 to about +500, about +1 to about +490, about +1 to about +480, about +1 to about +470, about +1 to about +460, about +1 to about +450, about +1 to about +440, about +1 to about +430, about +1 to about +420, about +1 to about +410, about +1 to about +400, about +1 to about +390, about +1 to about +380, about +1 to about +370, about +1 to about +360, about +1 to about +350, about +1 to about +340, about +1 to about +330, about +1 to about +320, about +1 to about +310, about +1 to about +300, about +1 to about +290, about +1 to about +280, about +1 to about +270, about +1 to about +260, about +1 to about +250, about +1 to about +240, about +1 to about +230, about +1 to about +220, about +1 to about +210, about +1 to about +200, about +1 to about +190, about +1 to about +180, about +1 to about +170, about +1 to about +160, about +1 to about +150, about +1 to about +140, about +1 to about +130, about +1 to about +120, about +1 to about +110, about +1 to about +100, about +1 to about +90, about +1 to about +80, about +1 to about +70, about +1 to about +60, about+1 to about +50, about +1 to about +40, about +1 to about +30, or about +1 to about +20, or about +1 to about +10 relative to 3’ splice site of the included exon.

[0309] In some embodiments, the targeted portion of the OPA1 pre-mRNA, e.g., the OPA1 NMD exon-containing pre-mRNA, is within the region +100 relative to the 5’ splice site (3’ end) of the included exon to -100 relative to the 3’ splice site (5’ end) of the included exon. In some embodiments, the targeted portion of the OPA1 NMD exon-containing pre-mRNA is within the NMD exon. In some embodiments, the target portion of the OPA1 NMD exon-containing pre- mRNA comprises a pseudo-exon and intron boundary.

[0310] The ASOs may be of any length suitable for specific binding and effective enhancement of splicing. In some embodiments, the ASOs consist of 8 to 50 nucleobases. For example, the ASO may be 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 40, 45, or 50 nucleobases in length. In some embodiments, the ASOs consist of more than 50 nucleobases. In some embodiments, the ASO is from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases,10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, 12 to 15 nucleobases, 13 to 50 nucleobases, 13 to 40 nucleobases, 13 to 35 nucleobases, 13 to 30nucleobases, 13 to 25 nucleobases, 13 to 20 nucleobases, 14 to 50 nucleobases, 14 to 40 nucleobases, 14 to 35 nucleobases, 14 to 30 nucleobases, 14 to 25 nucleobases, 14 to 20 nucleobases, 15 to 50 nucleobases, 15 to 40 nucleobases, 15 to 35 nucleobases, 15 to 30 nucleobases, 15 to 25 nucleobases, 15 to 20 nucleobases, 20 to 50 nucleobases, 20 to 40 nucleobases, 20 to 35 nucleobases, 20 to 30 nucleobases, 20 to 25 nucleobases, 25 to 50 nucleobases, 25 to 40 nucleobases, 25 to 35 nucleobases, or 25 to 30 nucleobases in length. In some embodiments, the ASOs are 18 nucleotides in length. In some embodiments, the ASOs are 15 nucleotides in length. In some embodiments, the ASOs are 25 nucleotides in length.

[0311] In some embodiments, two or more ASOs with different chemistries but complementary to the same targeted portion of the pre-mRNA, e.g., NMD exon-containing pre- mRNA, are used. In some embodiments, two or more ASOs that are complementary to different targeted portions of the pre-mRNA, e.g., the NMD exon-containing pre-mRNA, are used.

[0312] In some embodiments, the antisense oligonucleotides of the disclosure are chemically linked to one or more moieties or conjugates, e.g., a targeting moiety or other conjugate that enhances the activity or cellular uptake of the oligonucleotide. Such moieties include, but are not limited to, a lipid moiety, e.g., as a cholesterol moiety, a cholesteryl moiety, an aliphatic chain, e.g., dodecandiol or undecyl residues, a polyamine or a polyethylene glycol chain, or adamantane acetic acid. Oligonucleotides comprising lipophilic moieties and preparation methods have been described in the published literature. In embodiments, the antisense oligonucleotide is conjugated with a moiety including, but not limited to, an abasic nucleotide, a polyether, a polyamine, a polyamide, a peptides, a carbohydrate, e.g., N-acetylgalactosamine (GalNAc), N-Ac-Glucosamine (GluNAc), or mannose (e.g., mannose-6-phosphate), a lipid, or a polyhydrocarbon compound. Conjugates can be linked to one or more of any nucleotides comprising the antisense oligonucleotide at any of several positions on the sugar, base or phosphate group, as understood in the art and described in the literature, e.g., using a linker. Linkers can include a bivalent or trivalent branched linker. In embodiments, the conjugate is attached to the 3’ end of the antisense oligonucleotide. Methods of preparing oligonucleotide conjugates are described, e.g., in U.S. Pat. No. 8,450,467, “Carbohydrate conjugates as delivery agents for oligonucleotides,” incorporated by reference herein.

[0313] In some embodiments, the nucleic acid to be targeted by an ASO is an OPA1 pre- mRNA, e.g., NMD exon-containing pre-mRNA expressed in a cell, such as a eukaryotic cell. In some embodiments, the term “cell” may refer to a population of cells. In some embodiments, the cell is in a subject. In some embodiments, the cell is isolated from a subject. In some embodiments, the cell is ex vivo. In some embodiments, the cell is a condition or disease-relevant cell or a cell line. In some embodiments, the cell is in vitro (e.g., in cell culture).Pharmaceutical Compositions

[0314] Pharmaceutical compositions or formulations comprising the agent, e.g., antisense oligonucleotide, of the described compositions and for use in any of the described methods can be prepared according to conventional techniques well known in the pharmaceutical industry and described in the published literature. In embodiments, a pharmaceutical composition or formulation for treating a subject comprises an effective amount of any antisense oligomer as described herein, or a pharmaceutically acceptable salt, solvate, hydrate or ester thereof. The pharmaceutical formulation comprising an antisense oligomer may further comprise a pharmaceutically acceptable excipient, diluent or carrier.

[0315] Pharmaceutically acceptable salts are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio. (See, e.g., S. M. Berge, etal., J. Pharmaceutical Sciences, 66: 1-19 (1977), incorporated herein by reference for this purpose. The salts can be prepared in situ during the final isolation and purification of the compounds, or separately by reacting the free base form with a suitable organic acid. Examples of pharmaceutically acceptable, nontoxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other documented methodologies such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, di gluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2- hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3 -phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and aryl sulfonate.

[0316] In some embodiments, the compositions are formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. In embodiments, the compositions are formulated as suspensions inaqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances that increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and / or dextran. The suspension may also contain stabilizers. In embodiments, a pharmaceutical formulation or composition of the present disclosure includes, but is not limited to, a solution, emulsion, microemulsion, foam or liposome-containing formulation (e.g., cationic or noncationic liposomes).

[0317] The pharmaceutical composition or formulation described herein may comprise one or more penetration enhancers, carriers, excipients or other active or inactive ingredients as appropriate and well known to those of skill in the art or described in the published literature. In embodiments, liposomes also include sterically stabilized liposomes, e.g., liposomes comprising one or more specialized lipids. These specialized lipids result in liposomes with enhanced circulation lifetimes. In embodiments, a sterically stabilized liposome comprises one or more glycolipids or is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety. In some embodiments, a surfactant is included in the pharmaceutical formulation or compositions. The use of surfactants in drug products, formulations and emulsions is well known in the art. In embodiments, the present disclosure employs a penetration enhancer to effect the efficient delivery of the antisense oligonucleotide, e.g., to aid diffusion across cell membranes and / or enhance the permeability of a lipophilic drug. In some embodiments, the penetration enhancers are a surfactant, fatty acid, bile salt, chelating agent, or non-chelating nonsurfactant.

[0318] In some embodiments, the pharmaceutical formulation comprises multiple antisense oligonucleotides. In embodiments, the antisense oligonucleotide is administered in combination with another drug or therapeutic agent.

[0319] In some embodiments, the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer.

[0320] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL toabout 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0321] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0322] In some embodiments, the concentrate is a phosphate-buffered solution.

[0323] In some embodiments, the pharmaceutical formulation comprises: (a) an antisense oligomer, wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and (b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution at a concentration of about 1 mg / mL to about 200 mg / mL.

[0324] In some embodiments, the pharmaceutical formulation comprises (a) an antisense oligomer, wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and (b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, and wherein the antisense oligomer is a compound according to any one of the following chemical structures:

[0325] In some embodiments, the pharmaceutical formulation comprises (a) an antisense oligomer, wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and (b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, and wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0326] In some embodiments, the antisense oligomer is present in the solution at a concentration of about 1 mg / mL to about 200 mg / mL.

[0327] In some embodiments, the antisense oligomer is present in the solution at a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

[0328] In some embodiments, the antisense oligomer is present in the solution at a concentration of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0329] In some embodiments, the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer. In some embodiments, the pharmaceutical composition is prepared by diluting a concentrate consisting of the antisense oligomer.

[0330] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0331] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0332] In some embodiments, the concentrate is phosphate buffered. In some embodiments, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection. In some embodiments, the antisense oligomer is solubilized or diluted in a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection. In some embodiments, the antisense oligomer is solubilized or diluted in an isotonic solution. In some embodiments, the antisense oligomer is solubilized or diluted in a phosphate-buffered solution with at least pH 5.8. In some embodiments, the antisense oligomer is solubilized or diluted in a phosphate-buffered (pH 6.6 - 7.6) solution. In some embodiments, the pharmaceutical formulation does not comprise a preservative. In some embodiments, the pharmaceutical formulation is suitable for an intravitreal injection. In some embodiments, the pharmaceutical formulation is packaged in a single use vial.

[0333] In some embodiments, the pharmaceutical compositions are assembled into a kit comprising: (i) a concentrate comprising an antisense oligomer (ASO), wherein the ASO comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and (ii) a diluent, wherein the concentrate is miscible with the diluent; and (iii) instructions for diluting the concentrate with the diluent.

[0334] In some embodiments, the pharmaceutical compositions are assembled into a kit consisting of: (i) a concentrate comprising an antisense oligomer (ASO), wherein the ASO consists of a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and (ii) a diluent, wherein the concentrate is miscible with the diluent; and (iii) instructions for diluting the concentrate with the diluent.

[0335] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

[0336] In some embodiments, the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

[0337] In some embodiments, the concentrate is phosphate buffered. In some embodiments, the diluent is a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection. In some embodiments, the diluent is a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection. In some embodiments, the diluent comprises an isotonic solution. In some embodiments, the diluent comprises a phosphate-buffered solution with at least pH 5.8. In some embodiments, the diluent comprises a phosphate-buffered (pH 6.6 - 7.6) solution. In some embodiments, the concentrate or the diluent does not comprise a preservative.

[0338] In some embodiments, the diluent is a solution consisting of one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection. In some embodiments, the diluent is a solution consisting of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection. In some embodiments, the diluent consists of an isotonic solution. In some embodiments, the diluent consists of a phosphate- buffered solution with at least pH 5.8. In some embodiments, the diluent consists of a phosphate- buffered (pH 6.6 - 7.6) solution. In some embodiments, the concentrate or the diluent does not consist of a preservative.

[0339] In some embodiments, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 2 mg / mL to 200 mg / mL in the diluent.

[0340] In some embodiments, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL in the diluent.

[0341] In some embodiments, the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the antisense oligomer to a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL in the diluent.

[0342] In some embodiments, the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299.

[0343] In some embodiments, the antisense oligomer consists of a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299.

[0344] In some embodiments, the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292.

[0345] In some embodiments, the antisense oligomer consists of a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292.

[0346] In some embodiments, the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage. In some embodiments, the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N-methyl-acetamide (2’- NMA), or a 2’-O-methoxyethyl moiety. In some embodiments, the antisense oligomer comprises at least one modified sugar moiety. In some embodiments, each sugar moiety is a modified sugar moiety. In some embodiments, the antisense oligomer comprises a 5 ’-methylcytosine (5’-MeC). In some embodiments, each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’-MeC). In some embodiments, the antisense oligomer comprises a 5 ’-methyluracil (5’-MeU).

[0347] In some embodiments, the antisense oligomer consists of a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage. In some embodiments, the antisense oligomer consists of a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N-methyl-acetamide (2’- NMA), or a 2’-O-methoxyethyl moiety. In some embodiments, the antisense oligomer consists of at least one modified sugar moiety. In some embodiments, each sugar moiety is a modified sugar moiety. In some embodiments, the antisense oligomer consists of a 5 ’-methylcytosine (5’-MeC). In some embodiments, each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’-MeC). In some embodiments, the antisense oligomer consists of a 5 ’-methyluracil (5’-MeU).

[0348] In some embodiments, each cytosine or thymidine of the antisense oligomer is a 5 ’-methyluracil (5’-MeU). In some embodiments, the antisense oligomer consists of a phosphorothioate linkage. In some embodiments, each internucleoside linkage of the ASO is a phosphorothioate linkage. In some embodiments, the antisense oligomer consists of a locked nucleic acid (LNA).

[0349] In some embodiments, the antisense oligomer comprises from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases,9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases.

[0350] In some embodiments, the antisense oligomer consists of from 8 to 50 nucleobases,8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20- I l l -nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases,9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases.

[0351] In some embodiments, the antisense oligomer has any one of the following chemical structures:

[0352] In some embodiments, the antisense oligomer has any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

[0353] Described herein, in some aspects, are uses of an antisense oligomer for the manufacture of a medicament for treating a disease or condition characterized by a reduced expression or function of OPA1 protein in a human subject in need thereof, or reducing likelihood of the subject developing the disease or condition, wherein the medicament is administered to one eye of the subject at a dose of about 0.005 mg to about 20 mg, and wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299.

[0354] Described herein, in some aspects, are uses of an antisense oligomer for the manufacture of a medicament for treating a disease or condition characterized by a reduced expression or function of OPA1 protein in a human subject in need thereof, or reducing likelihood of the subject developing the disease or condition, wherein the medicament is administered to one eye of the subject at a dose of about 0.005 mg to about 20 mg, and wherein the antisense oligomer consists of a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299.

[0355] In some embodiments, the medicament is administered to the one eye of the subject at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mg to about 15 mg, about 0.01 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

[0356] In some embodiments, the medicament is administered to the one eye of the subject at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

[0357] In some embodiments, the medicament is administered to the one eye of the subject at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about 1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

[0358] In some embodiments, the medicament is administered to the one eye of the subject at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer. In some cases, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg to about 1.2 mg.Combination Therapies

[0359] In some embodiments, the ASOs disclosed in the present disclosure can be used in combination with one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents can comprise a small molecule. For example, the one or more additional therapeutic agents can comprise a small molecule described in WO2016128343A1,WO2017053982A1, WO2016196386A1, WO201428459A1, WO201524876 A2,WO2013119916 A2, and WO2014209841A2, which are incorporated by reference herein in their entirety. In some embodiments, the one or more additional therapeutic agents comprise an ASO that can be used to correct intron retention.Treatment of Subjects

[0360] Any of the compositions provided herein may be administered to an individual. “Individual” may be used interchangeably with “subject” or “patient.” An individual may be a mammal, for example, a human or animal such as a non-human primate, a rodent, a rabbit, a rat, a mouse, a horse, a donkey, a goat, a cat, a dog, a cow, a pig, or a sheep. In embodiments, the individual is a human. In embodiments, the individual is a fetus, an embryo, or a child. In other embodiments, the individual may be another eukaryotic organism, such as a plant. In some embodiments, the compositions provided herein are administered to a cell ex vivo.

[0361] In some embodiments, the compositions provided herein are administered to an individual as a method of treating a disease or disorder. In some embodiments, the individual has a genetic disease, such as any of the diseases described herein. In some embodiments, the individual is at risk of having a disease, such as any of the diseases described herein. In some embodiments, the individual is at increased risk of having a disease or disorder caused by insufficient amount of a protein or insufficient activity of a protein. If an individual is “at an increased risk” of having a disease or disorder caused insufficient amount of a protein or insufficient activity of a protein, the method involves preventative or prophylactic treatment. For example, an individual may be at an increased risk of having such a disease or disorder because of family history of the disease. Typically, individuals at an increased risk of having such a disease or disorder benefit from prophylactic treatment (e.g., by preventing or delaying the onset or progression of the disease or disorder). In embodiments, a fetus is treated in utero, e.g., by administering the ASO composition to the fetus directly or indirectly (e.g., via the mother).

[0362] In some cases, the subject pharmaceutical composition and method are applicable for treatment of a condition or disease associated with OPA1 deficiency. In some cases, the subject pharmaceutical composition and method are applicable for treatment of an eye disease or condition. In some cases, the subject pharmaceutical composition and method are applicable for treatment of Optic atrophy type 1, autosomal dominant optic atrophy (ADO A), ADOA-plus syndrome; a mitochondrial disorder; glaucoma; normal tension glaucoma; Charcot-Mari e-Tooth disease; mitochondria dysfunction; diabetic retinopathy; age-related macular degeneration; retinal ganglion cell death; mitochondrial fission-mediated mitochondrial dysfunction; progressive external ophthalmoplegia; deafness; ataxia; motor neuropathy; sensory neuropathy; myopathy; Behr syndrome; brain dysfunction; encephalopathy; peripheral neuropathy; fatal infantilemitochondrial encephalomyopathy; hypertrophic cardiomyopathy; spastic ataxic syndrome; sensory motor peripheral neuropathy; hypotonia; gastrointestinal dysmotility and dysphagia; optic atrophy; optic atrophy plus syndrome; Mitochondrial DNA depletion syndrome 14; late-onset cardiomyopathy; diabetic cardiomyopathy; Alzheimer’s Disease; focal segmental glomerulosclerosis; kidney disease; Huntington’s Disease; cognitive function decline in healthy aging; Prion diseases; late onset dementia and parkinsonism; mitochondrial myopathy; Leigh syndrome; Friedreich’s ataxia; Parkinson’s disease; MELAS (Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes); pyruvate dehydrogenase complex deficiency; chronic kidney disease; Leber’s hereditary optic neuropathy; obesity; age-related systemic neurodegeneration; skeletal muscle atrophy; heart and brain ischemic damage; or massive liver apoptosis.

[0363] In some embodiments, the disease or condition comprises Optic atrophy type 1. In some embodiments, the disease or condition comprises autosomal dominant optic atrophy (ADOA).

[0364] In some embodiments, the subject is characterized by having: (i) a heterozygous OPA1 gene variant; (ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments; (iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity; (iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or (v) any combination of (i)-(iv).

[0365] In some embodiments, the subject is additionally characterized by: (1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OP Al gene; (2) not having only benign or likely benign variants in the OP Al gene; (3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus); (4) not having been diagnosed with Behr syndrome; (5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases; (6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy; (7) not having or having a history of any ocular condition in either eye; (8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering; (9) not having a history of retinal photocoagulation; (10) not having a history or presence of retinal vein occlusion; (11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non- infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering; (12) not having dry age-related macular degeneration in either eye; (13) not having high myopia (>6 diopters); (14) not having a history of cancer (excepta diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated); (15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy; (16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery; (17) any combination of (1 )-(l 6).

[0366] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition and comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

[0367] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition and comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 90% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

[0368] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition and comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 100% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

[0369] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mgto about 15 mg, about 0.01 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

[0370] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

[0371] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about 1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

[0372] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer. Insome cases, the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg to about 1.2 mg.

[0373] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL to about 250 pL, about 10 pL to about 250 pL, about 20 pL to about 250 pL, about 30 pL to about 250 pL, about 40 pL to about 250 pL, about 50 pL to about 250 pL, about 60 pL to about 250 pL, about 70 pL to about 250 pL, about 80 pL to about 250 pL, about 100 pL to about 250 pL, about 120 pL to about 250 pL, about 150 pL to about 250 pL, about 160 pL to about 250 pL, about 180 pL to about 500 pL, about 200 pL to about 250 pL, about 220 pL to about 250 pL, about 5 pL to about 220 pL, about 10 pL to about 220 pL, about 20 pL to about 220 pL, about 30 pL to about 220 pL, about 40 pL to about 220 pL, about 50 pL to about 220 pL, about 60 pL to about 220 pL, about 70 pL to about 220 pL, about 80 pL to about 220 pL, about 100 pL to about 220 pL, about 120 pL to about 220 pL, about 150 pL to about 220 pL, about 160 pL to about 220 pL, about 180 pL to about 220 pL, about 5 pL to about 200 pL, about 10 pL to about 200 pL, about 20 pL to about 200 pL, about 30 pL to about 200 pL, about 40 pL to about 200 pL, about 50 pL to about 200 pL, about 60 pL to about 200 pL, about 70 pL to about 200 pL, about 80 pL to about 200 pL, about 100 pL to about 200 pL, about 120 pL to about 200 pL, about 150 pL to about 200 pL, about 160 pL to about 200 pL, about 180 pL to about 200 pL, about 5 pL to about 180 pL, about 10 pL to about 180 pL, about 20 pL to about 180 pL, about 30 pL to about 180 pL, about 40 pL to about 180 pL, about 50 pL to about 180 pL, about 60 pL to about 180 pL, about 70 pL to about 180 pL, about 80 pL to about 180 pL, about 100 pL to about 180 pL, about 120 pL to about 180 pL, about 150 pL to about 180 pL, about 5 pL to about 150 pL, about 10 pL to about 150 pL, about 20 pL to about 150 pL, about 30 pL to about 150 pL, about 40 pL to about 150 pL, about 50 pL to about 150 pL, about 60 pL to about 150 pL, about 70 pL to about 150 pL, about 80 pL to about 150 pL, about 100 pL to about 150 pL, about 120 pL to about 150 pL, about 5 pL to about 150 pL, about 10 pL to about 120 pL, about 20 pL to about 120 pL, about 30 pL to about 120 pL, about 40 pL to about 120 pL, about 50 pL to about 120 pL, about 60 pL to about 120 pL, about 70 pL to about 120 pL, about 80 pL to about 120 pL, about 100 pL to about 120 pL, about 5 pL to about 100 pL, about 10 pL to about 100 pL, about 20 pL to about 100 pL, about 30 pL to about 100 pL, about 40 pL to about 100 pL, about 50 pL to about 100 pL, about 60 pL to about 100 pL, about 70 pL to about 100 pL, about 80 pL to about 100 pL, about 5 pL to about 80 pL, about 10 pL to about 80 pL, about 20 pL to about 80 pL, about 30 pL to about 80 pL, about 40 pL to about 80 pL, about 50 pL to about 80 pL, about 60 pL to about 80 pL, about 5 pL to about 60 pL, about 10 pL to about 60 pL, about 20 pL to about 60 pL, about 30 pL to about 60 pL, about 40 pL to about 60 pL, or about 50 pL to about 60 pL.

[0374] In some embodiments, the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL, about 8 pL, about 10 pL, about 12 pL, about 15 pL, about 18 pL, about 20 pL, about 25 pL, about 28 pL, about 30 pL, about 35 pL, about 40 pL, about 45 pL, about 48 pL, about 50 pL, about 55 pL, about 60 pL, about 65 pL, about 70 pL, about 75 pL, about 80 pL, about 90 pL, about 100 pL, about 120 pL, about 150 pL, about 160 pL, about 180 pL, about 200 pL, about 220 pL, or about 250 pL. In some cases, the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 50 pL.

[0375] In some embodiments, the method comprises administering the pharmaceutical composition to both left eye and right eye of the subject. In some embodiments, the method comprises administering the pharmaceutical composition at the same dose to both the left eye and the right eye of the subject. In some embodiments, the method comprises administering the pharmaceutical composition at different doses to the left eye and the right eye of the subject. In some embodiments, the method comprises administering the pharmaceutical composition to at least one eye of the subject.

[0376] In some embodiments, the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299. In some embodiments, the antisense oligomer consists of a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299.

[0377] In some embodiments, the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292. In some embodiments, the antisense oligomer consists of a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292.

[0378] In some embodiments, administering comprises administering multiple doses of the pharmaceutical composition to the human subject. In some embodiments, administering comprises administering a first dose of the pharmaceutical composition to the human subject and a subsequent dose of the pharmaceutical composition to the human subject. In some embodiments, the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is not tolerated. In some embodiments, the subsequent dose is the same as the previous dose following an indication that administration of the previous dose is tolerated. In some embodiments, the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is tolerated. In some embodiments, the subsequent dose is thesame as the previous dose following an indication that administration of the previous dose is effective. In some embodiments, the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is effective. In some embodiments, the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is not effective.

[0379] In some embodiments, the pharmaceutical composition is a liquid composition. In some embodiments, the method comprises administering the pharmaceutical composition as a bolus injection over 1 to 60 minutes, 1 to 50 minutes, 1 to 40 minutes, 1 to 30 minutes, 1 to 20 minutes, 1 to 10 minutes, 1 to 5 minutes, or 1 to 3 minutes.

[0380] In some embodiments, the method comprises administering the pharmaceutical composition as a bolus injection. In some embodiments, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection. In some embodiments, the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection. In some embodiments, the antisense oligomer is solubilized or diluted in an isotonic solution. In some embodiments, the antisense oligomer is solubilized or diluted in a phosphate-buffered solution with at least pH 5.8. In some embodiments, the antisense oligomer is solubilized or diluted in a phosphate-buffered (pH 6.6 - 7.6) solution. In some embodiments, the pharmaceutical formulation does not comprise a preservative.

[0381] In some embodiments, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL.

[0382] In some embodiments, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL, about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mLto about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

[0383] In some embodiments, the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL. Autosomal dominant optic atrophy (ADOA) is the most common inherited optic nerve disorder and is characterized by retinal ganglion cell loss. In some cases, 65-90% of ADOA cases are caused by mutations in one allele of the OPA1 gene. OPA1 gene encodes an OPA1 protein that is a mitochondrial GTPase, which can have a critical maintenance role in mitochondria structure and function. Most OPA1 mutations can lead to a haploinsufficiency, resulting in about a 50% decrease of normal OPA1 protein levels. Approximately 1 out of 30,000 people are affected globally with a higher incidence of ~1 out of 10,000 in Denmark due to a founder effect. ADOA can present within the first decade of life. 80% of ADOA patients are symptomatic before 10 years of age. The disease can cause progressive and irreversible vision loss and up to 46% of patients are registered as legally blind.

[0384] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequenceidentity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by having: (i) a heterozygous OPA1 gene variant; (ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments; (iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity; (iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or (v) any combination of (i)-(iv).

[0385] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 90% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by having: (i) a heterozygous OPA1 gene variant; (ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments; (iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity; (iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or (v) any combination of (i)-(iv).

[0386] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 100% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by having: (i) a heterozygous OPA1 gene variant; (ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments; (iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity; (iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or (v) any combination of (i)-(iv).

[0387] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject ischaracterized by: (1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene; (2) not having only benign or likely benign variants in the OPA1 gene; (3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus); (4) not having been diagnosed with Behr syndrome; (5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases; (6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy; (7) not having or having a history of any ocular condition in either eye; (8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering; (9) not having a history of retinal photocoagulation; (10) not having a history or presence of retinal vein occlusion; (11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering; (12) not having dry age-related macular degeneration in either eye; (13) not having high myopia (>6 diopters); (14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated); (15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy; (16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery; or (17) any combination of (l)-(l 6).

[0388] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 90% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by: (1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene; (2) not having only benign or likely benign variants in the OPA1 gene; (3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus); (4) not having been diagnosed with Behr syndrome; (5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases; (6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy; (7) not having or having a history of any ocular condition in either eye; (8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to theadministering; (9) not having a history of retinal photocoagulation; (10) not having a history or presence of retinal vein occlusion; (11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering; (12) not having dry age-related macular degeneration in either eye; (13) not having high myopia (>6 diopters); (14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated); (15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy; (16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery; or (17) any combination of (l)-(l 6).

[0389] In some embodiments, the method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition comprises administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 100% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by: (1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene; (2) not having only benign or likely benign variants in the OPA1 gene; (3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus); (4) not having been diagnosed with Behr syndrome; (5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases; (6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy; (7) not having or having a history of any ocular condition in either eye; (8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering; (9) not having a history of retinal photocoagulation; (10) not having a history or presence of retinal vein occlusion; (11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering; (12) not having dry age-related macular degeneration in either eye; (13) not having high myopia (>6 diopters); (14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated); (15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy; (16) not having any history of nutritional deficiency (including B12 and / orfolate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery; or (17) any combination of (l)-(l 6).

[0390] In some cases, a therapeutic agent comprises an oligonucleotide. In some cases, a therapeutic agent comprises a vector, e.g., a viral vector, expressing an oligonucleotide that binds to the targeted region of a pre-mRNA the encodes the target peptide sequence. The methods provided herein can be adapted to contacting a vector that encodes an agent, e.g., an oligonucleotide, to a cell, so that the agent binds to a pre-mRNA in the cell and modulates the processing of the pre-mRNA. In some cases, the viral vector comprises an adenoviral vector, adeno-associated viral (AAV) vector, lentiviral vector, Herpes Simplex Virus (HSV) viral vector, retroviral vector, or any applicable viral vector. In some cases, a therapeutic agent comprises a gene editing tool that is configured to modify a gene encoding the target peptide sequence such that a gene region that encodes the inefficient translation region is deleted. In some cases, a gene editing tool comprises vector, e.g., viral vector, for gene editing based on CRISPR-Cas9, TALEN, Zinc Finger, or other applicable technologies.

[0391] Suitable routes for administration of ASOs of the present disclosure may vary depending on cell type to which delivery of the ASOs is desired. Multiple tissues and organs are affected by ADOA, with the eye being the most significantly affected tissue. The ASOs of the present disclosure may be administered to patients parenterally, for example, by intravitreal injection, intrathecal injection, intracerebroventricular injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection.

[0392] In some embodiments, the pharmaceutical composition is administered via intravitreal injection. In some embodiments, the method further comprises administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent comprises a small molecule. In some embodiments, the additional therapeutic agent comprises an antisense oligomer. In some embodiments, the additional therapeutic agent comprises an ophthalmologic drug. In some embodiments, the additional therapeutic agent consists of an antisense oligomer. In some embodiments, the additional therapeutic agent consists of an ophthalmologic drug.

[0393] In embodiments, the antisense oligonucleotide is administered with one or more agents capable of promoting penetration of the subject antisense oligonucleotide across the bloodbrain barrier by any method known in the art. For example, delivery of agents by administration of an adenovirus vector to motor neurons in muscle tissue is described in U.S. Pat. No. 6,632,427, “Adenoviral-vector-mediated gene transfer into medullary motor neurons,” incorporated herein by reference. Delivery of vectors directly to the brain, e.g., the striatum, the thalamus, the hippocampus, or the substantia nigra, is described, e.g., in U.S. Pat. No. 6,756,523, “Adenovirusvectors for the transfer of foreign genes into cells of the central nervous system particularly in brain,” incorporated herein by reference.

[0394] In some embodiments, the antisense oligonucleotides are linked or conjugated with agents that provide desirable pharmaceutical or pharmacodynamic properties. In embodiments, the antisense oligonucleotide is coupled to a substance, known in the art to promote penetration or transport across the blood-brain barrier, e.g., an antibody to the transferrin receptor. In embodiments, the antisense oligonucleotide is linked with a viral vector, e.g., to render the antisense compound more effective or increase transport across the blood-brain barrier. In embodiments, osmotic blood brain barrier disruption is assisted by infusion of sugars, e.g., meso erythritol, xylitol, D(+) galactose, D(+) lactose, D(+) xylose, dulcitol, myo-inositol, L(-) fructose, D(-) mannitol, D(+) glucose, D(+) arabinose, D(-) arabinose, cellobiose, D(+) maltose, D(+) raffinose, L(+) rhamnose, D(+) melibiose, D(-) ribose, adonitol, D(+) arabitol, L(-) arabitol, D(+) fucose, L(-) fucose, D(-) lyxose, L(+) lyxose, and L(-) lyxose, or amino acids, e.g., glutamine, lysine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glycine, histidine, leucine, methionine, phenylalanine, proline, serine, threonine, tyrosine, valine, and taurine. Methods and materials for enhancing blood brain barrier penetration are described, e.g., in U.S. Pat. No. 9,193,969, “Compositions and methods for selective delivery of oligonucleotide molecules to specific neuron types,” U.S. Pat. No. 4,866,042, “Method for the delivery of genetic material across the blood brain barrier,” U.S. Pat. No. 6,294,520, “Material for passage through the bloodbrain barrier,” and U.S. Pat. No. 6,936,589, “Parenteral delivery systems,” each incorporated herein by reference.

[0395] In some embodiments, subjects treated using the methods and compositions are evaluated for improvement in condition using any methods known and described in the art.Methods of Identifying Additional ASOs that Induce Exon Skipping

[0396] Also within the scope of the present disclosure are methods for identifying or determining ASOs that induce exon skipping of an OPA1 NMD exon-containing pre-mRNA. For example, a method can comprise identifying or determining ASOs that induce pseudo-exon skipping of an OPA1 NMD exon-containing pre-mRNA. ASOs that specifically hybridize to different nucleotides within the target region of the pre-mRNA may be screened to identify or determine ASOs that improve the rate and / or extent of splicing of the target intron. In some embodiments, the ASO may block or interfere with the binding site(s) of a splicing repressor(s) / silencer. Any method known in the art may be used to identify (determine) an ASO that when hybridized to the target region of the exon results in the desired effect (e.g., pseudoexon skipping, protein or functional RNA production). These methods also can be used for identifying ASOs that induce exon skipping of the included exon by binding to a targeted regionin an intron flanking the included exon, or in a non-included exon. An example of a method that may be used is provided below.

[0397] A round of screening, referred to as an ASO “walk” may be performed using ASOs that have been designed to hybridize to a target region of a pre-mRNA. For example, the ASOs used in the ASO walk can be tiled every 5 nucleotides from approximately 100 nucleotides upstream of the 3’ splice site of the included exon (e.g., a portion of sequence of the exon located upstream of the target / included exon) to approximately 100 nucleotides downstream of the 3’ splice site of the target / included exon and / or from approximately 100 nucleotides upstream of the 5’ splice site of the included exon to approximately 100 nucleotides downstream of the 5’ splice site of the target / included exon (e.g., a portion of sequence of the exon located downstream of the target / included exon). For example, a first ASO of 15 nucleotides in length may be designed to specifically hybridize to nucleotides +6 to +20 relative to the 3’ splice site of the target / included exon. A second ASO may be designed to specifically hybridize to nucleotides +11 to +25 relative to the 3’ splice site of the target / included exon. ASOs are designed as such spanning the target region of the pre-mRNA. In embodiments, the ASOs can be tiled more closely, e.g., every 1, 2, 3, or 4 nucleotides. Further, the ASOs can be tiled from 100 nucleotides downstream of the 5’ splice site, to 100 nucleotides upstream of the 3’ splice site. In some embodiments, the ASOs can be tiled from about 1,160 nucleotides upstream of the 3’ splice site, to about 500 nucleotides downstream of the 5’ splice site. In some embodiments, the ASOs can be tiled from about 500 nucleotides upstream of the 3’ splice site, to about 1,920 nucleotides downstream of the 3’ splice site.

[0398] One or more ASOs, or a control ASO (an ASO with a scrambled sequence, sequence that is not expected to hybridize to the target region) are delivered, for example by transfection, into a disease-relevant cell line that expresses the target pre-mRNA (e.g., a NMD exon-containing pre-mRNA described herein). The exon skipping effects of each of the ASOs may be assessed by any method known in the art, for example, by reverse transcriptase (RT)-PCR using primers that span the splice junction, as described in Example 4. A reduction or absence of a longer RT-PCR product produced using the primers spanning the region containing the included exon (e.g., including the flanking exons of the NMD exon) in ASO-treated cells as compared to in control ASO-treated cells indicates that splicing of the target NMD exon has been enhanced. In some embodiments, the exon skipping efficiency (or the splicing efficiency to splice the intron containing the NMD exon), the ratio of spliced to unspliced pre-mRNA, the rate of splicing, or the extent of splicing may be improved using the ASOs described herein. The amount of protein or functional RNA that is encoded by the target pre-mRNA can also be assessed to determine whether each ASO achieved the desired effect (e.g., enhanced functional protein production). Any methodknown in the art for assessing and / or quantifying protein production, such as Western Blotting, flow cytometry, immunofluorescence microscopy, and ELISA, can be used.

[0399] A second round of screening, referred to as an ASO “micro-walk” may be performed using ASOs that have been designed to hybridize to a target region of a pre-mRNA. The ASOs used in the ASO micro-walk are tiled every 1 nucleotide to further refine the nucleotide acid sequence of the pre-mRNA that when hybridized with an ASO results in exon skipping (or enhanced splicing of NMD exon).

[0400] Regions defined by ASOs that promote splicing of the target intron are explored in greater detail by means of an ASO “micro-walk”, involving ASOs spaced in 1-nt steps, as well as longer ASOs, typically 18-25 nt.

[0401] As described for the ASO walk above, the ASO micro-walk is performed by delivering one or more ASOs, or a control ASO (an ASO with a scrambled sequence, sequence that is not expected to hybridize to the target region), for example, by transfection, into a diseaserelevant cell line that expresses the target pre-mRNA. The splicing-inducing effects of each of the ASOs may be assessed by any method known in the art, for example, by reverse transcriptase (RT)-PCR using primers that span the NMD exon, as described herein (see, e.g., Example 4). A reduction or absence of a longer RT-PCR product produced using the primers spanning the NMD exon in ASO-treated cells as compared to in control ASO-treated cells indicates that exon skipping (or splicing of the target intron containing an NMD exon) has been enhanced. In some embodiments, the exon skipping efficiency (or the splicing efficiency to splice the intron containing the NMD exon), the ratio of spliced to unspliced pre-mRNA, the rate of splicing, or the extent of splicing may be improved using the ASOs described herein. The amount of protein or functional RNA that is encoded by the target pre-mRNA can also be assessed to determine whether each ASO achieved the desired effect (e.g., enhanced functional protein production). Any method known in the art for assessing and / or quantifying protein production, such as Western Blotting, flow cytometry, immunofluorescence microscopy, and ELISA, can be used.

[0402] ASOs that when hybridized to a region of a pre-mRNA result in exon skipping (or enhanced splicing of the intron containing a NMD exon) and increased protein production may be tested in vivo using animal models, for example, transgenic mouse models in which the full-length human gene has been knocked-in or in humanized mouse models of disease. Suitable routes for administration of ASOs may vary depending on the disease and / or the cell types to which delivery of the ASOs is desired. ASOs may be administered, for example, by intravitreal injection, intrathecal injection, intracerebroventricular injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection. Following administration, the cells, tissues, and / or organs of the model animals may be assessed to determine the effect of the ASOtreatment by, for example, evaluating splicing (e.g., efficiency, rate, extent) and protein production by methods known in the art and described herein. The animal models may also be any phenotypic or behavioral indication of the disease or disease severity.

[0403] Also within the scope of the present disclosure is a method to identify or validate an NMD-inducing exon in the presence of an NMD inhibitor, for example, cycloheximide. An exemplary method is provided in Example 2.SPECIFIC EMBODIMENTS

[0404] Embodiment Al. A method of treating Optic atrophy type 1 in a subject in need thereof, by increasing the expression of a target protein or functional RNA by a cell of the subject, wherein the cell has an mRNA that contains a nonsense-mediated RNA decay-inducing exon (NMD exon mRNA), and wherein the NMD exon mRNA encodes the target protein or functional RNA, the method comprising contacting the cell of the subject with a therapeutic agent that binds to a targeted portion of the NMD exon mRNA encoding the target protein or functional RNA, whereby the nonsense-mediated RNA decay-inducing exon is excluded from the NMD exon mRNA encoding the target protein or functional RNA, thereby increasing the level of mRNA encoding the target protein or functional RNA, and increasing the expression of the target protein or functional RNA in the cell of the subject.

[0405] Embodiment A2. The method of embodiment Al, wherein the target protein is OPA1.

[0406] Embodiment A3. A method of increasing expression of OPA1 protein by a cell having an mRNA that contains a nonsense-mediated RNA decay-inducing exon (NMD exon mRNA) and encodes OP Al protein, the method comprising contacting the cell with an agent that binds to a targeted portion of the NMD exon mRNA encoding OPA1 protein, whereby the nonsense-mediated RNA decay-inducing exon is excluded from the NMD exon mRNA encoding OPA1 protein, thereby increasing the level of mRNA encoding OPA1 protein, and increasing the expression of OPA1 protein in the cell.

[0407] Embodiment A4. The method of any one of embodiments Al to A3, wherein the nonsense-mediated RNA decay -inducing exon is spliced out from the NMD exon mRNA encoding the target protein or functional RNA.

[0408] Embodiment A5. The method of any one of embodiments Al to A4, wherein the target protein does not comprise an amino acid sequence encoded by the nonsense-mediated RNA decay-inducing exon.

[0409] Embodiment A6. The method of any one of embodiments Al to A5, wherein the target protein is a full-length target protein.

[0410] Embodiment A7. The method of any one of embodiments Al to A6, wherein the agent is an antisense oligomer (ASO) complementary to the targeted portion of the NMD exon mRNA.

[0411] Embodiment A8. The method of any one of embodiments Al to A7, wherein the mRNA is pre-mRNA.

[0412] Embodiment A9. The method of any one of embodiments Al to A8, wherein the contacting comprises contacting the therapeutic agent to the mRNA, wherein the mRNA is in a nucleus of the cell.

[0413] Embodiment A10. The method of any one of embodiments Al to A9, wherein the target protein or the functional RNA corrects a deficiency in the target protein or functional RNA in the subject.

[0414] Embodiment Al 1. The method of any one of embodiments Al to A10, wherein the cells are in or from a subject with a condition caused by a deficient amount or activity of an OPA1 protein.

[0415] Embodiment A12. The method of any one of embodiments Al to Al l, wherein the deficient amount of the target protein is caused by haploinsufficiency of the target protein, wherein the subject has a first allele encoding a functional target protein, and a second allele from which the target protein is not produced, or a second allele encoding a nonfunctional target protein, and wherein the antisense oligomer binds to a targeted portion of a NMD exon mRNA transcribed from the first allele.

[0416] Embodiment Al 3. The method of any one of embodiments Al to Al 1, wherein the subject has a condition caused by a disorder resulting from a deficiency in the amount or function of the target protein, wherein the subj ect has a first mutant allele from which(i) the target protein is produced at a reduced level compared to production from a wildtype allele,(ii) the target protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the target protein is not produced, and(b) a second mutant allele from which(i) the target protein is produced at a reduced level compared to production from a wildtype allele,(ii) the target protein is produced in a form having reduced function compared to an equivalent wild-type protein, or(iii) the target protein is not produced, andwherein when the subject has a first mutant allele (a)(iii). , the second mutant allele is (b)(i) or (b)(ii) and wherein when the subject has a second mutant allele (b)(iii), the first mutant allele is (a)(i) or (a)(ii), and wherein the NMD exon mRNA is transcribed from either the first mutant allele that is (a)(i) or (a)(ii), and / or the second allele that is (b)(i) or (b)(ii).

[0417] Embodiment A14. The method of embodiment A13, wherein the target protein is produced in a form having reduced function compared to the equivalent wild-type protein.

[0418] Embodiment A15. The method of embodiment A13, wherein the target protein is produced in a form that is fully-functional compared to the equivalent wild-type protein.

[0419] Embodiment Al 6. The method of any one of embodiments Al to Al 5, wherein the targeted portion of the NMD exon mRNA is within the nonsense-mediated RNA decay-inducing exon.

[0420] Embodiment Al 7. The method of any one of embodiments Al to Al 5, wherein the targeted portion of the NMD exon mRNA is either upstream or downstream of the nonsense- mediated RNA decay-inducing exon.

[0421] Embodiment A18. The method of any one of embodiments Al to A17, wherein the NMD exon mRNA comprises a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 2 or 3.

[0422] Embodiment Al 9. The method of any one of embodiments Al to Al 7, wherein theNMD exon mRNA is encoded by a genetic sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 1.

[0423] Embodiment A20. The method of any one of embodiments Al to Al 7, wherein the targeted portion of the NMD exon mRNA comprises a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3.

[0424] Embodiment A21. The method of any one of embodiments Al to A20, wherein the agent is an antisense oligomer (ASO) and wherein the ASO comprises a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complementary to at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3.

[0425] Embodiment A22. The method of any one of embodiments Al to Al 5, wherein the targeted portion of the NMD exon mRNA is within the nonsense-mediated RNA decay-inducing exon 6x of OP Al, exon 7x of OP Al, or exon 28x of OP Al.

[0426] Embodiment A23. The method of any one of embodiments Al to Al 5, wherein the targeted portion of the NMD exon mRNA is upstream or downstream of the nonsense-mediated RNA decay-inducing exon 6x of OPA1, exon 7x of OPA1, or exon 28x of OPAl.

[0427] Embodiment A24. The method of any one of embodiments Al to Al 5, wherein the targeted portion of the NMD exon mRNA comprises an exon-intron junction exon 6x of OPA L exon 7x of OP Al, or exon 28x of OPAL

[0428] Embodiment A25. The method of any one of embodiments Al to A24, wherein the target protein produced is full-length protein, or wild-type protein.

[0429] Embodiment A26. The method of any one of embodiments Al to A25, wherein the total amount of the mRNA encoding the target protein or functional RNA produced in the cell contacted with the antisense oligomer is increased about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to the total amount of the mRNA encoding the target protein or functional RNA produced in a control cell.

[0430] Embodiment A27. The method of any one of embodiments Al to A25, wherein the total amount of the mRNA encoding the target protein or functional RNA produced in the cell contacted with the antisense oligomer is increased about 20% to about 300%, about 50% to about 300%, about 100% to about 300%, about 150% to about 300%, about 20% to about 50%, about 20% to about 100%, about 20% to about 150%, about 20% to about 200%, about 20% to about 250%, about 50% to about 100%, about 50% to about 150%, about 50% to about 200%, about 50% to about 250%, about 100% to about 150%, about 100% to about 200%, about 100% to about 250%, about 150% to about 200%, about 150% to about 250%, about 200% to about 250%, at least about 10%, at least about 20%, at least about 50%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, or at least about 300%, compared to the total amount of the mRNA encoding the target protein or functional RNA produced in a control cell.

[0431] Embodiment A28. The method of one any of embodiments Al to A25, wherein the total amount of target protein produced by the cell contacted with the antisense oligomer is increased about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5- fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9- fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1- fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to the total amount of target protein produced by a control cell.

[0432] Embodiment A29. The method of one any of embodiments Al to A25, wherein the total amount of target protein produced by the cell contacted with the antisense oligomer is increased about 20% to about 300%, about 50% to about 300%, about 100% to about 300%, about 150% to about 300%, about 20% to about 50%, about 20% to about 100%, about 20% to about 150%, about 20% to about 200%, about 20% to about 250%, about 50% to about 100%, about50% to about 150%, about 50% to about 200%, about 50% to about 250%, about 100% to about 150%, about 100% to about 200%, about 100% to about 250%, about 150% to about 200%, about 150% to about 250%, about 200% to about 250%, at least about 10%, at least about 20%, at least about 50%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, or at least about 300%, compared to the total amount of target protein produced by a control cell.

[0433] Embodiment A30. The method of any one of embodiments Al to A29, wherein the agent is an antisense oligomer (ASO) and wherein the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

[0434] Embodiment A31. The method of any one of embodiments Al to A30, wherein the agent is an antisense oligomer (ASO) and wherein the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl, a 2’-Fluoro, or a 2’ -O-m ethoxy ethyl moiety.

[0435] Embodiment A32. The method of any one of embodiments Al to A31, wherein the agent is an antisense oligomer (ASO) and wherein the antisense oligomer comprises at least one modified sugar moiety.

[0436] Embodiment A33. The method of embodiment A32, wherein each sugar moiety is a modified sugar moiety.

[0437] Embodiment A34. The method of any one of embodiments Al to A33, wherein the agent is an antisense oligomer (ASO) and wherein the antisense oligomer consists of from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases,8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases,10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases.

[0438] Embodiment A35. The method of any one of embodiments Al to A34, wherein the agent is an antisense oligomer (ASO) and wherein the antisense oligomer is at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, complementary to the targeted portion of the NMD exon mRNA encoding the protein.

[0439] Embodiment A36. The method of any one of embodiments Al to A35, wherein the method further comprises assessing OPA1 mRNA or protein expression.

[0440] Embodiment A37. The method of any one of embodiments Al to A36, wherein Optic atrophy type 1 is treated and wherein the antisense oligomer binds to a targeted portion of an OPA1 NMD exon mRNA, wherein the targeted portion is within SEQ ID NO: 2 or 3.

[0441] Embodiment A38. The method of any one of embodiments Al to A37, wherein the subject is a human.

[0442] Embodiment A39. The method of any one of embodiments Al to A38, wherein the subject is a non-human animal.

[0443] Embodiment A40. The method of any one of embodiments Al to A39, wherein the subject is a fetus, an embryo, or a child.

[0444] Embodiment A41. The method of any one of embodiments Al to A40, wherein the cells are ex vivo.

[0445] Embodiment A42. The method of any one of embodiments Al to A41, wherein the therapeutic agent is administered by intrathecal injection, intracerebroventricular injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection of the subj ect.

[0446] Embodiment A43. The method of any of embodiments Al to A42, wherein the method further comprises administering a second therapeutic agent to the subject.

[0447] Embodiment A44. The method of embodiment A43, wherein the second therapeutic agent is a small molecule.

[0448] Embodiment A45. The method of embodiment A43, wherein the second therapeutic agent is an ASO.

[0449] Embodiment A46. The method of any one of embodiments A43 to A45, wherein the second therapeutic agent corrects intron retention.

[0450] Embodiment A47. An antisense oligomer as used in a method of any of embodiments Al to A46.

[0451] Embodiment A48. An antisense oligomer comprising a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3.

[0452] Embodiment A49. A pharmaceutical composition comprising the antisense oligomer of embodiment A47 or A48 and an excipient.

[0453] Embodiment A50. A method of treating a subject in need thereof, comprising administering the pharmaceutical composition of embodiment A49 to the subject, wherein the administering is by intravitreal injection, intrathecal injection, intracerebroventricular injection, intraperitoneal injection, intramuscular injection, subcutaneous injection, or intravenous injection.

[0454] Embodiment A51. A composition comprising a therapeutic agent for use in a method of increasing expression of a target protein or a functional RNA by cells to treat Optic atrophy type 1 in a subject in need thereof, associated with a deficient protein or deficient functional RNA, wherein the deficient protein or deficient functional RNA is deficient in amount or activity in the subject, wherein the target protein is:(a) the deficient protein; or(b) a compensating protein which functionally augments or replaces the deficient protein or in the subj ect; and wherein the functional RNA is:(c) the deficient RNA; or(d) a compensating functional RNA which functionally augments or replaces the deficient functional RNA in the subject; wherein the therapeutic agent enhances exclusion of the nonsense-mediated RNA decay -inducing exon from the NMD exon mRNA encoding the target protein or functional RNA, thereby increasing production or activity of the target protein or the functional RNA in the subject.

[0455] Embodiment A52. A composition comprising a therapeutic agent for use in a method of treating a condition associated with OPA1 protein in a subject in need thereof, the method comprising the step of increasing expression of OPA1 protein by cells of the subject, wherein the cells have an mRNA that contains a nonsense-mediated RNA decay-inducing exon (NMD exon mRNA) and encodes OPA1 protein, the method comprising contacting the cells with the therapeutic agent, whereby the nonsense-mediated RNA decay-inducing exon is excluded from the NMD exon mRNA that encodes OPA1 protein, thereby increasing the level of mRNA encoding OPA1 protein, and increasing the expression of OPA1 protein in the cells of the subject.

[0456] Embodiment A53. The composition of embodiment A52, wherein the condition is a disease or disorder.

[0457] Embodiment A54. The composition of embodiment A53, wherein the disease or disorder is Optic atrophy type 1.

[0458] Embodiment A55. The composition of any one of embodiments A52 to 54, wherein the OPA1 protein and NMD exon mRNA are encoded by the OPA1 gene.

[0459] Embodiment A56. The composition of any one of embodiments A51 to A55, wherein the nonsense-mediated RNA decay -inducing exon is spliced out from the NMD exon mRNA encoding the OPA1 protein.

[0460] Embodiment A57. The composition of any one of embodiments A51 to A56, wherein the OPA1 protein does not comprise an amino acid sequence encoded by the nonsense- mediated RNA decay-inducing exon.

[0461] Embodiment A58. The composition of any one of embodiments A51 to A57, wherein the OPA1 protein is a full-length OPA1 protein.

[0462] Embodiment A59. The composition of any one of embodiments A51 to A58, wherein the therapeutic agent is an antisense oligomer (ASO) complementary to the targeted portion of the NMD exon mRNA.

[0463] Embodiment A60. The composition of any of embodiments A51 to A59, wherein the therapeutic agent is an antisense oligomer (ASO) and wherein the antisense oligomer targets a portion of the NMD exon mRNA that is within the nonsense-mediated RNA decay-inducing exon.

[0464] Embodiment A61. The composition of any of embodiments A51 to A59, wherein the therapeutic agent is an antisense oligomer (ASO) and wherein the antisense oligomer targets a portion of the NMD exon mRNA that is upstream or downstream of the nonsense-mediated RNA decay-inducing exon.

[0465] Embodiment A62. The composition of any one of embodiments A51 to A61, wherein the target protein is OPA1.

[0466] Embodiment A63. The composition of embodiment A62, wherein the NMD exon mRNA comprises a sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 2 or 3.

[0467] Embodiment A64. The composition of embodiment A62, wherein the NMD exon mRNA is encoded by a genetic sequence with at least about 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to SEQ ID NO: 1.

[0468] Embodiment A65. The composition of embodiment A62, wherein the targeted portion of the NMD exon mRNA comprises a sequence with at least 80%, 85%, 90%, 95%, 97%, or 100% sequence identity to a region comprising at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3.

[0469] Embodiment A66. The composition of any one of embodiments A62 to A65, wherein the targeted portion of the NMD exon mRNA is within the nonsense-mediated RNA decay-inducing exon 6x of OPA1, exon 7x of OPA1, or exon 28x of OPAL

[0470] Embodiment A67. The composition of any one of embodiments A62 to A65, wherein the targeted portion of the NMD exon mRNA is upstream or downstream of the nonsense- mediated RNA decay-inducing exon 6x of OPA1, exon 7x of OPA1, or exon 28x of OPAl.

[0471] Embodiment A68. The composition of any one of embodiments A62 to A65, wherein the targeted portion of the NMD exon mRNA comprises an exon-intron junction of exon 6x of OPAl, exon 7x of OPAl, or exon 28x of OPAl.

[0472] Embodiment A69. The composition of any one of embodiments A62 to A68, wherein the therapeutic agent is an antisense oligomer (ASO) and wherein the ASO comprises a sequence that is at least about 80%, 85%, 90%, 95%, 97%, or 100% complementary to a region comprising at least 8 contiguous nucleic acids of SEQ ID NO: 2 or 3.

[0473] Embodiment A70. The composition of any one of embodiments A51 to A69, wherein the mRNA encoding the target protein or functional RNA is a full-length mature mRNA, or a wild-type mature mRNA.

[0474] Embodiment A71. The composition of any one of embodiments A51 to A70, wherein the target protein produced is full-length protein, or wild-type protein.

[0475] Embodiment A72. The composition of any one of embodiments A51 to A71, wherein the therapeutic agent is an antisense oligomer (ASO) and wherein the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

[0476] Embodiment A73. The composition of any of embodiments A51 to A72, wherein the therapeutic agent is an antisense oligomer (ASO) and wherein said antisens...

Claims

CLAIMSWhat is claimed is:

1. A method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, and wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

2. A method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by having:(i) a heterozygous OPA1 gene variant;(ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments;(iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity;(iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or(v) any combination of (i)-(iv).

3. A method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer comprises a nucleotide sequence having at least 80% sequence identity to the sequence set forth in any one of SEQ ID NO: 6-275 or 280-299, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA), and wherein the subject is characterized by:(1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene;(2) not having only benign or likely benign variants in the OPA1 gene;(3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus);(4) not having been diagnosed with Behr syndrome;(5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases;(6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy;(7) not having or having a history of any ocular condition in either eye;(8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering;(9) not having a history of retinal photocoagulation;(10) not having a history or presence of retinal vein occlusion;(11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering;(12) not having dry age-related macular degeneration in either eye;(13) not having high myopia (>6 diopters);(14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated);(15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy;(16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B 12 or folate, not having had bariatric surgery; or(17) any combination of (1)-(16).

4. A method of treating a subject having a disease or condition or reducing likelihood of the subject developing the disease or condition, the method comprising administering to the subject a pharmaceutical composition comprising an antisense oligomer, wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

5. The method of any one of claims 2-4, wherein the method comprises administering to one eye of the subject the pharmaceutical composition at a dose of about 0.005 to about 20 mg of the antisense oligomer.

6. The method of any one of claims 1-5, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mg to about 15 mg, about 0.01 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

7. The method of any one of claims 1-5, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

8. The method of any one of claims 1-5, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about 1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

9. The method of any one of claims 1-5, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer.

10. The method of any one of claims 1-9, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL to about 250 pL, about 10 pL to about 250 pL, about 20 pL to about 250 pL, about 30 pL to about 250 pL, about 40 pL to about 250 pL, about 50 pL to about 250 pL, about 60 pL to about 250 pL, about 70 pL to about 250 pL, about 80 pL to about 250 pL, about 100 pL to about 250 pL, about 120 pL to about 250 pL, about 150 pL to about 250 pL, about 160 pL to about 250 pL, about 180 pL to about 500 pL, about 200 pL to about 250 pL, about 220 pL to about 250 pL, about 5 pL to about 220 pL, about 10 pL to about 220 pL, about 20 pL to about 220 pL, about 30 pL to about 220 pL, about 40 pL to about 220 pL, about 50 pL to about 220 pL, about 60 pL to about 220 pL, about 70 pL to about 220 pL, about 80 pL to about 220 pL, about 100 pL to about 220 pL, about 120 pL to about 220 pL, about 150 pL to about 220 pL, about 160 pL to about 220 pL, about 180 pL to about 220 pL, about 5 pL to about 200 pL, about 10 pL to about 200 pL, about 20 pL to about 200 pL, about 30 pL to about 200 pL, about 40 pL to about 200 pL, about 50 pL to about 200 pL, about 60 pL to about 200 pL, about 70 pL to about 200 pL, about 80 pL to about 200 pL, about 100 pL to about 200 pL, about 120 pL to about 200 pL, about 150 pL to about 200 pL, about 160pL to about 200 pL, about 180 pL to about 200 pL, about 5 pL to about 180 pL, about 10 pL to about 180 pL, about 20 pL to about 180 pL, about 30 pL to about 180 pL, about 40 pL to about 180 pL, about 50 pL to about 180 pL, about 60 pL to about 180 pL, about 70 pL to about 180 pL, about 80 pL to about 180 pL, about 100 pL to about 180 pL, about 120 pL to about 180 pL, about 150 pL to about 180 pL, about 5 pL to about 150 pL, about 10 pL to about 150 pL, about 20 pL to about 150 pL, about 30 pL to about 150 pL, about 40 pL to about 150 pL, about 50 pL to about 150 pL, about 60 pL to about 150 pL, about 70 pL to about 150 pL, about 80 pL to about 150 pL, about 100 pL to about 150 pL, about 120 pL to about 150 pL, about 5 pL to about 150 pL, about 10 pL to about 120 pL, about 20 pL to about 120 pL, about 30 pL to about 120 pL, about 40 pL to about 120 pL, about 50 pL to about 120 pL, about 60 pL to about 120 pL, about 70 pL to about 120 pL, about 80 pL to about 120 pL, about 100 pL to about 120 pL, about 5 pL to about 100 pL, about 10 pL to about 100 pL, about 20 pL to about 100 pL, about 30 pL to about 100 pL, about 40 pL to about 100 pL, about 50 pL to about 100 pL, about 60 pL to about 100 pL, about 70 pL to about 100 pL, about 80 pL to about 100 pL, about 5 pL to about 80 pL, about 10 pL to about 80 pL, about 20 pL to about 80 pL, about 30 pL to about 80 pL, about 40 pL to about 80 pL, about 50 pL to about 80 pL, about 60 pL to about 80 pL, about 5 pL to about 60 pL, about 10 JJ.L to about 60 pL, about 20 pL to about 60 pL, about 30 pL to about 60 pL, about 40 pL to about 60 pL, or about 50 pL to about 60 pL.

11. The method of any one of claims 1-9, wherein the method comprises administering to the one eye of the subject the pharmaceutical composition in a volume of about 5 pL, about 8 pL, about 10 pL, about 12 pL, about 15 pL, about 18 pL, about 20 pL, about 25 pL, about 28 pL, about 30 pL, about 35 pL, about 40 pL, about 45 pL, about 48 pL, about 50 pL, about 55 pL, about 60 pL, about 65 pL, about 70 pL, about 75 pL, about 80 pL, about 90 pL, about 100 pL, about 120 pL, about 150 pL, about 160 pL, about 180 pL, about 200 pL, about 220 pL, or about 250 pL.

12. The method of any one of claims 1-11, wherein the method comprises administering the pharmaceutical composition to both left eye and right eye of the subject.

13. The method of claim 12, wherein the method comprises administering the pharmaceutical composition at the same dose to both the left eye and the right eye of the subject.

14. The method of claim 12, wherein the method comprises administering the pharmaceutical composition at different doses to the left eye and the right eye of the subject.

15. The method of any one of claims 1-3 or 5-14, wherein the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to the sequence set forth in any one of SEQ ID NOs: 6-275 or 280-299.

16. The method of any one of claims 1-3 or 5-14, wherein the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to the sequence set forth in any one of SEQ ID NO: 36, 236, 242, 250, 280-283, 288, or 290-292.

17. The method of any one of claims 1-3 or 5-14, wherein the antisense oligomer modulates splicing of a non-sense mediated RNA decay-inducing exon (NMD exon) from a pre-mRNA in a cell of the subject, wherein the pre-mRNA encodes the OPA1 protein and comprises the NMD exon, thereby modulating a level of processed mRNA that is processed from the pre-mRNA, and modulating expression of the OPA1 protein in the cell.

18. The method of claim 17, wherein the antisense oligomer:(a) binds to a targeted portion of the pre-mRNA;(b) modulates binding of a factor involved in splicing of the NMD exon; or(c) a combination of (a) and (b).

19. The method of claim 18, wherein the targeted portion of the pre-mRNA is proximal to the NMD exon.

20. The method of claim 18, wherein the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of 5’ end of the NMD exon.

21. The method of claim 18, wherein the targeted portion of the pre-mRNA is at least about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides, about 40 nucleotides, about 30 nucleotides, about 20 nucleotides, about 10 nucleotides, about 5 nucleotides, about 4 nucleotides, about 2 nucleotides, about 1 nucleotides upstream of 5’ end of the NMD exon.

22. The method of claim 18, wherein the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of 3’ end of the NMD exon.

23. The method of claim 18, wherein the targeted portion of the pre-mRNA is at least about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60nucleotides, about 50 nucleotides, about 40 nucleotides, about 30 nucleotides, about 20 nucleotides, about 10 nucleotides, about 5 nucleotides, about 4 nucleotides, about 2 nucleotides, about 1 nucleotides downstream of 3’ end of the NMD exon.

24. The method of claim 18, wherein the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193628509.

25. The method of claim 18, wherein the targeted portion of the pre-mRNA is about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides upstream of genomic site GRCh38 / hg38: chr3 193628509.

26. The method of claim 18, wherein the targeted portion of the pre-mRNA is at most about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193628616.

27. The method of claim 18, wherein the targeted portion of the pre-mRNA is about 1500 nucleotides, about 1000 nucleotides, about 800 nucleotides, about 700 nucleotides, about 600 nucleotides, about 500 nucleotides, about 400 nucleotides, about 300 nucleotides, about 200 nucleotides, about 100 nucleotides, about 80 nucleotides, about 70 nucleotides, about 60 nucleotides, about 50 nucleotides downstream of genomic site GRCh38 / hg38: chr3 193628616.

28. The method of claim 18, wherein the targeted portion of the pre-mRNA is located in an intronic region between two canonical exonic regions of the pre-mRNA, and wherein the intronic region contains the NMD exon.

29. The method of claim 18, wherein the targeted portion of the pre-mRNA at least partially overlaps with the NMD exon.

30. The method of claim 18, wherein the targeted portion of the pre-mRNA at least partially overlaps with an intron upstream or downstream of the NMD exon.

31. The method of claim 18, wherein the targeted portion of the pre-mRNA comprises 5 ’ NMD exon-intron junction or 3’ NMD exon-intron junction.

32. The method of claim 18, wherein the targeted portion of the pre-mRNA is within the NMD exon.

33. The method of claim 18, wherein the targeted portion of the pre-mRNA comprises about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more consecutive nucleotides of the NMD exon.

34. The method of any one of claims 17-33, wherein the NMD exon comprises a sequence with at least 80%, at least 90%, or 100% sequence identity to SEQ ID NO: 279.

35. The method of any one of claims 17-33, wherein the NMD exon comprises a sequence of SEQ ID NO: 279.

36. The method of claim 18, wherein the targeted portion of the pre-mRNA is within the nonsense mediated RNA decay-inducing exon GRCh38 / hg38: chr3 193628509 to 193628616.

37. The method of claim 18, wherein the targeted portion of the pre-mRNA is upstream or downstream of the non-sense mediated RNA decay -inducing exon GRCh38 / hg38: chr3 193628509 to 193628616.

38. The method of claim 18, wherein the targeted portion of the pre-mRNA comprises an exonintronjunction of exon GRCh38 / hg38: chr3 193628509 to 193628616.

39. The method of any one of claims 17-38, wherein the OPA1 protein expressed from the processed mRNA is a full-length OPA1 protein or a wild-type OPA1 protein.

40. The method of any one of claims 17-38, wherein the OPA1 protein expressed from the processed mRNA is a functional OPA1 protein.

41. The method of any one of claims 17-38, wherein the OPA1 protein expressed from the processed mRNA is at least partially functional as compared to a wild-type OPA1 protein.

42. The method of any one of claims 17-38, wherein the OPA1 protein expressed from the processed mRNA is at least partially functional as compared to a full-length wild-type OPA1 protein.

43. The method of any one of claims 17-42, wherein the antisense oligomer promotes exclusion of the NMD exon from the pre-mRNA.

44. The method of claim 43, wherein the exclusion of the NMD exon from the pre-mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5- fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

45. The method of any one of claims 17-44, wherein the method results in an increase in the level of the processed mRNA in the cell.

46. The method of claim 45, wherein the level of the processed mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10-fold, about 1.1 to about 5- fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1-fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5- fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

47. The method of any one of claims 17-46, wherein the method results in an increase in the expression of the 0PA1 protein in the cell.

48. The method of claim 47, wherein a level of the 0PA1 protein expressed from the processed mRNA in the cell contacted with the antisense oligomer is increased by about 1.1 to about 10-fold, about 1.5 to about 10-fold, about 2 to about 10-fold, about 3 to about 10-fold, about 4 to about 10- fold, about 1.1 to about 5-fold, about 1.1 to about 6-fold, about 1.1 to about 7-fold, about 1.1 to about 8-fold, about 1.1 to about 9-fold, about 2 to about 5-fold, about 2 to about 6-fold, about 2 to about 7-fold, about 2 to about 8-fold, about 2 to about 9-fold, about 3 to about 6-fold, about 3 to about 7-fold, about 3 to about 8-fold, about 3 to about 9-fold, about 4 to about 7-fold, about 4 to about 8-fold, about 4 to about 9-fold, at least about 1.1 -fold, at least about 1.5-fold, at least about 2-fold, at least about 2.5-fold, at least about 3-fold, at least about 3.5-fold, at least about 4-fold, at least about 5-fold, or at least about 10-fold, compared to in the absence of the antisense oligomer.

49. The method of any one of claims 1-3 or 5-48, wherein the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

50. The method of any one of claims 1-3 or 5-48, wherein the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N-methyl-acetamide (2’-NMA), or a 2’-O-methoxyethyl moiety.

51. The method of any one of claims 1-3 or 5-48, wherein the antisense oligomer comprises at least one modified sugar moiety.

52. The method of claim 51, wherein each sugar moiety is a modified sugar moiety.

53. The method of any one of claims 1-3 or 5-52, wherein the antisense oligomer comprises a 5 ’-methyl cytosine (5’-MeC).

54. The method of any one of claims 1-3 or 5-53, wherein each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’-MeC).

55. The method of any one of claims 1-3 or 5-54, wherein the antisense oligomer comprises a 5 ’-methyluracil (5’-MeU).

56. The method of any one of claims 1-3 or 5-55, wherein the antisense oligomer comprises a phosphorothioate linkage.

57. The method of any one of claims 1-3 or 5-56, wherein each internucleoside linkage of the ASO is a phosphorothioate linkage.

58. The method of any one of claims 1-3 or 5-57, wherein the antisense oligomer comprises a locked nucleic acid (LNA).

59. The method of any one of claims 1-3 or 5-58, wherein the antisense oligomer is from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases, 9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases,10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases in length.

60. The method of any one of claims 1-3 or 5-59, wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

61. The method of claim 4 or 60, wherein the antisense oligomer is a compound according to any of the following structures:

62. The method of any one of claims 1-61, wherein the pharmaceutical composition is a liquid composition.

63. The method of any one of claims 1-62, wherein the method comprises administering the pharmaceutical composition as a bolus injection.

64. The method of any one of claims 1-63, wherein the method comprises administering the pharmaceutical composition as a bolus injection over 1 to 60 minutes, 1 to 50 minutes, 1 to 40 minutes, 1 to 30 minutes, 1 to 20 minutes, 1 to 10 minutes, 1 to 5 minutes, or 1 to 3 minutes.

65. The method of any one of claims 1-64, wherein the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection.

66. The method of any one of claims 1-64, wherein the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection.

67. The method of any one of claims 1-66, wherein the antisense oligomer is solubilized or diluted in an isotonic solution.

68. The method of any one of claims 1-67, wherein the antisense oligomer is solubilized or diluted in a phosphate-buffered solution with at least pH 5.8.

69. The method of any one of claims 1-67, wherein the antisense oligomer is solubilized or diluted in a phosphate-buffered (pH 6.6 - 7.6) solution.

70. The method of any one of claims 1-69, wherein the pharmaceutical formulation does not comprise a preservative.

71. The method of any one of claims 1-70, wherein the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL.

72. The method of any one of claims 1-70, wherein the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL to about 200 mg / mL, about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

73. The method of any one of claims 1-70, wherein the antisense oligomer is present in the pharmaceutical composition at a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

74. The method of any one of claims 1-73, wherein the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer.

75. The method of claim 74, wherein the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

76. The method of claim 74, wherein the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

77. The method of any one of claims 74-76, wherein the concentrate is a phosphate-buffered solution.

78. The method of any one of claims 1-77, wherein the subject is a human subject.

79. The method of any one of claims 1 or 4-78, wherein the disease or condition is associated with a deficient amount or activity of the OPA1 protein.

80. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises an eye disease or condition.

81. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises a cardiovascular disease or condition.

82. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises a neurological disease or condition.

83. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises ADOA-plus syndrome; a mitochondrial disorder; glaucoma; normal tension glaucoma; Charcot- Marie-Tooth disease; mitochondria dysfunction; diabetic retinopathy; age-related macular degeneration; retinal ganglion cell death; mitochondrial fission-mediated mitochondrial dysfunction; progressive external ophthalmoplegia; deafness; ataxia; motor neuropathy; sensoryneuropathy; myopathy; Behr syndrome; brain dysfunction; encephalopathy; peripheral neuropathy; fatal infantile mitochondrial encephalomyopathy; hypertrophic cardiomyopathy; spastic ataxic syndrome; sensory motor peripheral neuropathy; hypotonia; gastrointestinal dysmotility and dysphagia; optic atrophy; optic atrophy plus syndrome; Mitochondrial DNA depletion syndrome 14; late-onset cardiomyopathy; diabetic cardiomyopathy; Alzheimer’s Disease; focal segmental glomerulosclerosis; kidney disease; Huntington’s Disease; cognitive function decline in healthy aging; Prion diseases; late onset dementia and parkinsonism; mitochondrial myopathy; Leigh syndrome; Friedreich’s ataxia; Parkinson’s disease; MELAS (Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes); pyruvate dehydrogenase complex deficiency; chronic kidney disease; Leber’s hereditary optic neuropathy; obesity; age-related systemic neurodegeneration; skeletal muscle atrophy; heart and brain ischemic damage; or massive liver apoptosis.

84. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises Optic atrophy type 1.

85. The method of any one of claims 1 or 4-79, wherein the disease or condition comprises autosomal dominant optic atrophy (ADOA).

86. The method of claim 85, wherein the subject is characterized by having:(i) a heterozygous 0PA1 gene variant;(ii) clear ocular media to allow for adequate visualization of the vitreous and fundus and to achieve appropriate quality of all ophthalmic assessments;(iii) an AREDS Clinical Lens Standard of <1 for posterior subcapsular (PSC) opacity;(iv) a BCVA EDTRS letter score of >35 and <70 with each eye individually that is administered with the pharmaceutical composition; or(v) any combination of (i)-(iv).

87. The method of claim 86, wherein the subject is additionally characterized by:(1) not having a gain-of-function variant, or compound heterozygous or homozygous pathogenic or likely pathogenic variant in OPA1 gene;(2) not having only benign or likely benign variants in the OPA1 gene;(3) not having extraocular phenotypic manifestations of (syndromic) ADOA (ADOA-plus);(4) not having been diagnosed with Behr syndrome;(5) not having a known pathogenic mutation in another gene implicated in optic atrophy or retinal diseases;(6) not having diabetic retinopathy with potential for development of proliferative diabetic retinopathy, diabetic macular edema, or optic neuropathy;(7) not having or having a history of any ocular condition in either eye;(8) not having a history of intraocular surgery or corneal surgery including refractive surgery in either eye within 12 weeks prior to the administering;(9) not having a history of retinal photocoagulation;(10) not having a history or presence of retinal vein occlusion;(11) not being considered to be at risk for uveitis or ocular infection because of having any of the following: an active flare of non-infectious uveitis or an episode of infectious uveitis or other ocular infection in either eye within 12 months prior to the administering;(12) not having dry age-related macular degeneration in either eye;(13) not having high myopia (>6 diopters);(14) not having a history of cancer (except a diagnosis of basal cell, squamous cell skin cancer, or carcinoma in situ of the cervix that has been successfully treated);(15) not taking, or having taken at any time, any medication or treatment that can or might cause an optic neuropathy;(16) not having any history of nutritional deficiency (including B12 and / or folate deficiencies), or not having a known deficiency in serum B12 or folate, not having had bariatric surgery;(17) any combination of (l)-(l 6).

88. The method of any one of claims 1-87, wherein administering comprises administering multiple doses of the pharmaceutical composition to the human subject.

89. The method of claim 88, wherein administering comprises administering a first dose of the pharmaceutical composition to the human subject and a subsequent dose of the pharmaceutical composition to the human subject.

90. The method of claim 89, wherein the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is not tolerated.

91. The method of claim 89, wherein the subsequent dose is the same as the previous dose following an indication that administration of the previous dose is tolerated.

92. The method of claim 89, wherein the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is tolerated.

93. The method of claim 89, wherein the subsequent dose is the same as the previous dose following an indication that administration of the previous dose is effective.

94. The method of claim 89, wherein the subsequent dose is lower than the previous dose following an indication that administration of the previous dose is effective.

95. The method of claim 89, wherein the subsequent dose is higher than the previous dose following an indication that administration of the previous dose is not effective.

96. The method of any one of claims 1-95, wherein the pharmaceutical composition is administered via intracerebroventricular injection, intraperitoneal injection, intramuscular injection, intrathecal injection, subcutaneous injection, oral administration, synovial injection, intravitreal administration, subretinal injection, topical application, implantation, or intravenous injection.

97. The method of any one of claims 1-87, wherein the pharmaceutical composition is administered via intravitreal injection.

98. The method of any one of claims 1-97, wherein the method further comprises administering an additional therapeutic agent.

99. The method of claim 98, wherein the additional therapeutic agent comprises a small molecule.

100. The method of claim 98, wherein the additional therapeutic agent comprises an antisense oligomer.

101. The method of claim 98, wherein the additional therapeutic agent comprises an ophthalmologic drug.

102. An antisense oligomer that is a compound according to any one of the following structures:or a pharmaceutically acceptable salt thereof.

103. An antisense oligomer having any of the following structures:

104. A pharmaceutical formulation comprising:(a) an antisense oligomer, wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution at a concentration of about 1 mg / mL to about 200 mg / mL.

105. A pharmaceutical formulation comprising:(a) an antisense oligomer; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, and wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

106. A pharmaceutical formulation comprising:(a) an antisense oligomer; and(b) a pharmaceutically acceptable diluent; wherein the antisense oligomer is dissolved or suspended in a solution, and wherein the antisense oligomer has any one of the following chemical structures:

107. The pharmaceutical formulation of claim 105 or 106, wherein the antisense oligomer is present in the solution at a concentration of about 1 mg / mL to about 200 mg / mL.

108. The pharmaceutical formulation of claim 104 or 107, wherein the antisense oligomer is present in the solution at a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL toabout 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL.

109. The pharmaceutical formulation of claim 104, wherein the antisense oligomer is present in the solution at a concentration of about 1 mg / mL, about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14 mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

110. The pharmaceutical formulation of any one of claims 104-109, wherein the pharmaceutical composition is prepared by diluting a concentrate comprising the antisense oligomer.

111. The pharmaceutical formulation of claim 110, wherein the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

112. The pharmaceutical composition of claim 110, wherein the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

113. The pharmaceutical formulation of any one of claims 104-112, wherein the concentrate is phosphate buffered.

114. The pharmaceutical formulation of any one of claims 104-113, wherein the antisense oligomer is solubilized or diluted in a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection.

115. The pharmaceutical formulation of any one of claims 104-113, wherein the antisense oligomer is solubilized or diluted in a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection.

116. The pharmaceutical formulation of any one of claims 104-113, wherein the antisense oligomer is solubilized or diluted in an isotonic solution.

117. The pharmaceutical formulation of any one of claims 104-116, wherein the antisense oligomer is solubilized or diluted in a phosphate-buffered solution with at least pH 5.8.

118. The pharmaceutical formulation of any one of claims 104-116, wherein the antisense oligomer is solubilized or diluted in a phosphate-buffered (pH 6.6 - 7.6) solution.

119. The pharmaceutical formulation of any one of claims 104- 118, wherein the pharmaceutical formulation does not comprise a preservative.

120. The pharmaceutical formulation of any one of claims 104-119, wherein the pharmaceutical formulation is suitable for an intravitreal injection.

121. The pharmaceutical formulation of any one of claims 104-120, wherein the pharmaceutical formulation is packaged in a single-use vial.

122. A kit compri sing :(i) a concentrate comprising an antisense oligomer (ASO), wherein the ASO comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299; and(ii) a diluent, wherein the concentrate is miscible with the diluent; and(iii) instructions for diluting the concentrate with the diluent.

123. The kit of claim 122, wherein the antisense oligomer is present in the concentrate at a concentration of about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, or about 40 mg / mL to about 60 mg / mL.

124. The kit of claim 122, wherein the antisense oligomer is present in the concentrate at a concentration of about 40 mg / mL, about 45 mg / mL, about 50 mg / mL, about 55 mg / mL, about 60 mg / mL, about 65 mg / mL, about 70 mg / mL, about 80 mg / mL, about 90 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL.

125. The kit of any one of claims 122-124, wherein the concentrate is phosphate buffered.

126. The kit of any one of claims 122-125, wherein the diluent is a solution comprising one or more of sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, or water for injection.

127. The kit of any one of claims 122-125, wherein the diluent is a solution comprising sodium chloride, sodium phosphate dibasic, potassium phosphate monobasic, and water for injection.

128. The kit of any one of claims 122-127, wherein the diluent comprises an isotonic solution.

129. The kit of any one of claims 122-128, wherein the diluent comprises a phosphate-buffered solution with at least pH 5.8.

130. The kit of any one of claims 122-128, wherein the diluent comprises a phosphate-buffered (pH 6.6 - 7.6) solution.

131. The kit of any one of claims 122-130, wherein the concentrate or the diluent does not comprise a preservative.

132. The kit of any one of claims 122-131, wherein the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 2 mg / mL to 200 mg / mL in the diluent.

133. The kit of any one of claims 122-132, wherein the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the ASO to a concentration of about 5 mg / mL to about 200 mg / mL, about 10 mg / mL to about 200 mg / mL, about 15 mg / mL to about 200 mg / mL, about 20 mg / mL to about 200 mg / mL, about 25 mg / mL to about 200 mg / mL, about 30 mg / mL to about 200 mg / mL, about 35 mg / mL to about 200 mg / mL, about 40 mg / mL to about 200 mg / mL, about 50 mg / mL to about 200 mg / mL, about 60 mg / mL to about 200 mg / mL, about 80 mg / mL to about 200 mg / mL, about 100 mg / mL to about 200 mg / mL, about 150 mg / mL to about 200 mg / mL, about 180 mg / mL to about 200 mg / mL, 2 mg / mL to about 150 mg / mL, about 5 mg / mL to about 150 mg / mL, about 10 mg / mL to about 150 mg / mL, about 15 mg / mL to about 150 mg / mL, about 20 mg / mL to about 150 mg / mL, about 25 mg / mL to about 150 mg / mL, about 30 mg / mL to about 150 mg / mL, about 35 mg / mL to about 150 mg / mL, about 40 mg / mL to about 150 mg / mL, about 50 mg / mL to about 150 mg / mL, about 60 mg / mL to about 150 mg / mL, about 80 mg / mL to about 150 mg / mL, about 100 mg / mL to about 150 mg / mL, 2 mg / mL to about 100 mg / mL, about 5 mg / mL to about 100 mg / mL, about 10 mg / mL to about 100 mg / mL, about 15 mg / mL to about 100 mg / mL, about 20 mg / mL to about 100 mg / mL, about 25 mg / mL to about 100 mg / mL, about 30 mg / mL to about 100 mg / mL, about 35 mg / mL to about 100 mg / mL, about 40 mg / mL to about 100 mg / mL, about 50 mg / mL to about 100 mg / mL, about 60 mg / mL to about 100 mg / mL, about 80 mg / mL to about 100 mg / mL, 2 mg / mL to about 80 mg / mL, about 5 mg / mL to about 80 mg / mL, about 10 mg / mL to about 80 mg / mL, about 15 mg / mL to about 80 mg / mL, about 20 mg / mL to about 80 mg / mL, about 25 mg / mL to about 80 mg / mL, about 30 mg / mL to about 80 mg / mL, about 35 mg / mL to about 80 mg / mL, about 40 mg / mL to about 80 mg / mL, about 60 mg / mL to about 80 mg / mL, 2 mg / mL to about 60 mg / mL, about 5 mg / mL to about 60 mg / mL, about 10 mg / mL to about 60 mg / mL, about 15 mg / mL to about 60 mg / mL, about 20 mg / mL to about 60 mg / mL, about 25 mg / mL to about 60 mg / mL, about 30 mg / mL to about 60 mg / mL, about 35 mg / mL to about 60 mg / mL, about 40 mg / mL to about 60 mg / mL, 2 mg / mL to about 40 mg / mL, about 5 mg / mL to about 40 mg / mL, about 10 mg / mL to about 40 mg / mL, about 15 mg / mL to about 40 mg / mL, about 20 mg / mL to about 40 mg / mL, or about 25 mg / mL to about 40 mg / mL in the diluent.

134. The kit of any one of claims 122-132, wherein the instructions for diluting the concentrate with the diluent comprise instructions for diluting or solubilizing the antisense oligomer to a concentration of about 2 mg / mL, about 3 mg / mL, about 4 mg / mL, about 5 mg / mL, about 6 mg / mL, about 7 mg / mL, about 8 mg / mL, about 9 mg / mL, about 10 mg / mL, about 12 mg / mL, about 14mg / mL, about 15 mg / mL, about 16 mg / mL, about 18 mg / mL, about 20 mg / mL, about 22 mg / mL, about 24 mg / mL, about 26 mg / mL, about 28 mg / mL, about 30 mg / mL, about 35 mg / mL, about 40 mg / mL, about 50 mg / mL, about 60 mg / mL, about 80 mg / mL, about 100 mg / mL, about 120 mg / mL, about 140 mg / mL, about 160 mg / mL, about 180 mg / mL, or about 200 mg / mL in the diluent.

135. The pharmaceutical composition or kit of any one of claims 104 or 108-134, wherein the antisense oligomer comprises a nucleotide sequence having at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 6-275 or 280-299.

136. The pharmaceutical composition or kit of any one of claims 104 or 108-134, wherein the antisense oligomer comprises a nucleotide sequence having at least 80%, at least 90%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 36, 236, 242, 250, 280-283, 288, and 290-292.

137. The pharmaceutical composition or kit of any one of claims 104 or 108-136, wherein the antisense oligomer comprises a backbone modification comprising a phosphorothioate linkage or a phosphorodiamidate linkage.

138. The pharmaceutical composition or kit of any one of claims 104 or 108-137, wherein the antisense oligomer comprises a phosphorodiamidate morpholino, a locked nucleic acid, a peptide nucleic acid, a 2’-O-methyl moiety, a 2’-Fluoro moiety, a 2’-O-N-methyl-acetamide (2’-NMA), or a 2’-O-methoxyethyl moiety.

139. The pharmaceutical composition or kit of any one of claims 104 or 108-138, wherein the antisense oligomer comprises at least one modified sugar moiety.

140. The pharmaceutical composition or kit of claim 139, wherein each sugar moiety is a modified sugar moiety.

141. The pharmaceutical composition or kit of any one of claims 104 or 108-140, wherein the antisense oligomer comprises a 5 ’-methylcytosine (5’-MeC).

142. The pharmaceutical composition or kit of any one of claims 104 or 108-141, wherein each cytosine of the antisense oligomer is a 5 ’-methylcytosine (5’-MeC).

143. The pharmaceutical composition or kit of any one of claims 104 or 108-142, wherein the antisense oligomer comprises a 5 ’-methyluracil (5’-MeU).

144. The pharmaceutical composition or kit of any one of claims 104 or 108-143, wherein the antisense oligomer comprises a phosphorothioate linkage.

145. The pharmaceutical composition or kit of any one of claims 104 or 108-144, wherein each internucleoside linkage of the ASO is a phosphorothioate linkage.

146. The pharmaceutical composition or kit of any one of claims 104 or 108-145, wherein the antisense oligomer comprises a locked nucleic acid (LNA).

147. The pharmaceutical composition or kit of any one of claims 104 or 108-146, wherein the antisense oligomer is from 8 to 50 nucleobases, 8 to 40 nucleobases, 8 to 35 nucleobases, 8 to 30 nucleobases, 8 to 25 nucleobases, 8 to 20 nucleobases, 8 to 15 nucleobases, 9 to 50 nucleobases,9 to 40 nucleobases, 9 to 35 nucleobases, 9 to 30 nucleobases, 9 to 25 nucleobases, 9 to 20 nucleobases, 9 to 15 nucleobases, 10 to 50 nucleobases, 10 to 40 nucleobases, 10 to 35 nucleobases, 10 to 30 nucleobases, 10 to 25 nucleobases, 10 to 20 nucleobases, 10 to 15 nucleobases, 11 to 50 nucleobases, 11 to 40 nucleobases, 11 to 35 nucleobases, 11 to 30 nucleobases, 11 to 25 nucleobases, 11 to 20 nucleobases, 11 to 15 nucleobases, 12 to 50 nucleobases, 12 to 40 nucleobases, 12 to 35 nucleobases, 12 to 30 nucleobases, 12 to 25 nucleobases, 12 to 20 nucleobases, or 12 to 15 nucleobases in length.

148. The pharmaceutical composition or kit of any one of claims 104 or 108-134, wherein the antisense oligomer is a compound according to any one of the following chemical structures:or a pharmaceutically acceptable salt thereof.

149. The pharmaceutical composition or kit of any one of claims 104 or 108-134, wherein the antisense oligomer has any one of the following chemical structures:

150. Use of an antisense oligomer for the manufacture of a medicament for treating a disease or condition characterized by a reduced expression or function of OPA1 protein in a human subject in need thereof, or reducing likelihood of the subject developing the disease or condition, wherein the medicament is administered to one eye of the subject at a dose of about 0.005 mg to about 20 mg, and wherein the antisense oligomer comprises a sequence with at least 80% sequence identity to any one of SEQ ID NOs: 6-275 or 280-299.

151. The use of claim 150, wherein the medicament is administered to the one eye of the subj ect at a dose of about 0.005 mg to about 15 mg, about 0.005 mg to about 10 mg, about 0.005 mg to about 5 mg, about 0.005 mg to about 1 mg, about 0.01 mg to about 15 mg, about 0.01 mg to about 10 mg, about 0.01 mg to about 5 mg, about 0.01 mg to about 2.5 mg, about 0.01 mg to about 1.0 mg, about 0.01 mg to about 0.5 mg, about 0.01 mg to about 0.25 mg, about 0.01 mg to about 0.1 mg, about 0.01 mg to about 0.05 mg, about 0.05 mg to about 10 mg, about 0.05 mg to about 5 mg, about 0.05 mg to about 2.5 mg, about 0.05 mg to about 1.0 mg, about 0.05 mg to about 0.5 mg, about 0.05 mg to about 0.25 mg, about 0.05 mg to about 0.1 mg, about 0.1 mg to about 5 mg, about 0.1 mg to about 2.5 mg, about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.5 mg, or about 0.1 mg to about 0.25 mg of the antisense oligomer.

152. The use of claim 150, wherein the medicament is administered to the one eye of the subject at a dose of about 0.005 mg, about 0.01 mg, about 0.05 mg, about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, about 1.5 mg, about 1.75 mg, about 2.0 mg, about 2.25 mg, about 2.5 mg, about 2.75 mg, about 3 mg, about 3.5 mg, about 4.0 mg, about 4.5 mg, about 5.0 mg, about 5.5 mg, about 6.0 mg, about 7.0 mg, about 8.0 mg, about 9.0 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of the antisense oligomer.

153. The use of claim 150, wherein the medicament is administered to the one eye of the subj ect at a dose of about 0.1 mg to about 1.5 mg, about 0.1 mg to about 1.4 mg, about 0.1 mg to about 1.2 mg, about 0.1 mg to about 1.0 mg, about 0.1 mg to about 0.8 mg, about 0.1 mg to about 0.7 mg, about 0.1 mg to about 0.5 mg, about 0.1 mg to about 0.3 mg, about 0.2 mg to about 1.5 mg, about 0.2 mg to about 1.4 mg, about 0.2 mg to about 1.2 mg, about 0.2 mg to about 1.0 mg, about 0.2 mg to about 0.8 mg, about 0.2 mg to about 0.7 mg, about 0.2 mg to about 0.5 mg, about 0.3 mg to about 1.5 mg, about 0.3 mg to about 1.4 mg, about 0.3 mg to about 1.2 mg, about 0.3 mg to about 1.0 mg, about 0.3 mg to about 0.8 mg, about 0.3 mg to about 0.7 mg, about 0.3 mg to about 0.5 mg, about 0.5 mg to about 1.5 mg, about 0.5 mg to about 1.4 mg, about 0.5 mg to about 1.2 mg, about 0.5 mg to about 1.0 mg, about 0.5 mg to about 0.8 mg, about 0.5 mg to about 0.7 mg, about 0.7 mg to about 1.5 mg, about 0.7 mg to about 1.4 mg, about 0.7 mg to about 1.2 mg, about 0.7 mg to about 1.0 mg, about 0.8 mg to about 1.5 mg, about 0.8 mg to about 1.4 mg, about 0.8 mg to about 1.2 mg, about 0.8 mg to about 1.0 mg, about 1.0 mg to about 1.5 mg, about 1.0 mg to about 1.4 mg, about 1.0 mg to about 1.2 mg, about 1.2 mg to about 1.5 mg, or about 1.2 mg to about 1.4 mg of the antisense oligomer.

154. The use of claim 150, wherein the medicament is administered to the one eye of the subject at a dose of about 0.1 mg, about 0.2 mg, about 0.3 mg, 0.4 mg, about 0.5 mg, about 0.6 mg, about 0.7 mg, about 0.8 mg, about 0.9 mg, about 1.0 mg, about 1.1 mg, about 1.2 mg, about 1.3 mg, about 1.4 mg, or about 1.5 mg of the antisense oligomer.

Images