Sirna compositions and methods for usp10 modulation

Abstract

Nucleic acid complexes that induce RNA interference and related therapeutics are described.

Description

[0001] SIRNA COMPOSITIONS AND METHODS FOR USP10 MODULATION

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 63 / 561 ,859, filed on March 6, 2024, the entire contents of which are incorporated herein.

[0004] FIELD OF THE DISCLOSURE

[0005] Nucleic acid complexes that induce RNA interference and related therapeutics are described.

[0006] GOVERNMENT INTEREST

[0007] This invention was made with government support under grant NIH EY024942, and grant NIH R44 EY035188, awarded by the National Institutes of Health. The government has certain rights in the invention.

[0008] DESCRIPTION OF THE TEXT FILE SUBMITTED ELECTRONICALLY

[0009] The instant application contains a sequence listing, which has been submitted in XML format via EFS-Web. The contents of the XML copy named “137434-5001 _Sequence_Listing”, was created on March 5, 2025 and is 42,202 bytes in size, the contents of which are incorporated herein by reference in their entirety.

[0010] BACKGROUND

[0011] Oligonucleotide-based gene silencing modalities, such as RNA interference (RNAi) or antisense oligonucleotides (ASO), are viewed as an exciting therapeutic opportunity. However, RNA-based therapeutics face several challenges. For example, RNAi modalities are difficult to deliver in high enough amounts to the target tissue due to clearance by the real system and degradation by nucleases in tissues and in the blood stream. In addition, toxicity due to off-target effects and activation of the immune system are also hurdles for RNA-based therapeutics. Thus, there is a need for novel RNA-based compositions and related therapeutics that are stable and efficient at target delivery.

[0012] SUMMARY Accordingly the present disclosure describes a composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, and the sense strand is about 16 to about 18 nucleotides in length and has sequence complementarity to the antisense strand. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. In embodiments, the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10. In embodiments, the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

[0013] In embodiments, the sense strand is about 16 nucleotides in length, or is about 17 nucleotides in length, or is about 18 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 16 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 17 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 18 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, at least 16, 17, 18, 19, or 20 nucleotides of the antisense strand are complementary to the target.

[0014] In embodiments, the sense strand comprises the base sequence of SEQ ID NOs : 1 -2, or a variant thereof. In embodiments, the antisense strand comprises the base sequence of SEQ ID NO: 3 or a variant thereof. In embodiments, the variant comprises about 1, or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0015] In embodiments, the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 6 , or a variant thereof. In embodiments, the antisense strand consists of the base sequence of SEQ ID NO: 7 , or a variant thereof. In embodiments, the nucleic acid complex comprises one or more chemical modifications. In embodiments, the nucleic acid complex comprises one or more chemical modifications selected from Table 2.

[0016] In embodiments, the chemical modification is a 2'-O-methylated nucleoside, a phosphorothioate bond, or a hydrophobic moiety. In embodiments, the chemical modification is selected from a locked nucleic acid (LNA), phosphorothioate, 2'-O-Methyl, 2'-O-Methoxyethyl, 2'-O-alkyl-RNA unit, 2'-0Me-RNA unit, 2 -amino-DNA unit, 2'-fluoro-DNA unit, peptide nucleic acid (PNA) unit, hexitol nucleic acids (HNA) unit, INA unit, and a 2'- O-(2-Methoxyethyl)-RNA (2' MOE RNA) unit. In embodiments, the chemical modification is a 2' fluoro group. In embodiments, the chemical modification is 2'-O-methyl (2'0Me) group. In embodiments, the chemical modification is a phosphorothioate linker. In embodiments, the chemical modification is a 5'- vinylphosphonate 2'0-mU group. In embodiments, the chemical modification is a cholesterol conjugate, or the chemical modification comprises docosanoic acid (DCA).

[0017] In embodiments, the composition comprises a modified sequence selected from: (a) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 9) 5'

[0018] [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 10) 5'

[0019] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][P s][mC][Ps][fG] 3'; (b) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 11) 5' [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 12) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][Ps][mC][ Ps][fG] 3'; (c) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 13) 5' [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fll][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 14) 5'

[0020] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][P s][mC][Ps][fG] 3'; and (d) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 15) 5' [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 16) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][Ps][mC][ Ps][fG] 3'; wherein f represents 2' Fluoro, m represents 2' O-methyl, Ps represents phosphorothioate linker, 5Phos represents 5' phosphate, 3'CholTEG represents Cholesterol conjugate, and vP-mll represents 5'- Vinylphosphonate 2'0-mll.

[0021] In embodiments, disclosed herein is a composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, the sense strand is about 15 nucleotides in length and having sequence complementarity to the antisense strand; and the antisense strand and the sense strand comprise one or more chemical modifications, selected from P, #, f, 2’ Fluor, m, 2’0-methyl, Choi 3’, cholesterol and docosanoic acid (DCA), wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0022] In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. In embodiments, the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

[0023] In embodiments, the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10. In embodiments, the nucleic acid complex has: a sense strand consisting of 15 nucleotides in length and an antisense strand consisting of 20 nucleotides in length.

[0024] In embodiments, the sense strand comprises the base sequence of SEQ ID NO: 5 or a variant thereof. In embodiments, the antisense strand comprises the base sequence of SEQ ID NO: 3 , or a variant thereof. In embodiments, the variant comprises about 1, or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0025] In embodiments, the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof. In embodiments, the antisense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof. In embodiments, the nucleic acid complex comprises one or more chemical modifications. In embodiments, the nucleic acid complex comprises one or more chemical modifications selected from Table 3.

[0026] In embodiments, the composition comprises a modified sequence selected from a nucleic acid complex comprising: a sense strand selected from

[0027] 5' mC#mA#mG.mA.mC.mC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 17);

[0028] 5' mC#mA#mG.mG.mA.mA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 18);

[0029] 5' mT#mA#mT.mT.mC.mT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 19);

[0030] 5' mC#mA#mG.mA.fC.fC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 20);

[0031] 5' mC#mA#mG.mG.fA.fA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 21);

[0032] 5' mT#mA#mT.mT.fC.fT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 22);

[0033] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 23);

[0034] 5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 24);

[0035] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 25);

[0036] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.fA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 26);

[0037] 5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.fT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 27);

[0038] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.fT.mA.mA#mT#mA-Chol 31(SEQ ID NO: 28);

[0039] 5' mC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#mA-Chol 3' (SEQ ID NO: 29);

[0040] 5' mC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#mA-Chol 3' (SEQ ID NO: 30);

[0041] 5' mT#mA#fT.mT.fC.mT.fT.mG.fG.mT.fT.mA.fA#mT#mA-Chol 3' (SEQ ID NO: 31);

[0042] 5' fC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#fA-Chol 3' (SEQ ID NO: 32);

[0043] 5' fC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#fA-Chol 3' (SEQ ID NO: 33);

[0044] 5' fT#mA#fT.mT.fC.mT.fT.mG.fG.mT.fT.mA.fA#mT#fA-Chol 3' (SEQ ID NO: 34); and an antisense strand of:

[0045] 5' PmT#fC#mT.mT.mT.mC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 35);

[0046] 5' mT#fA#mT.mC.mA.mG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 36);

[0047] 5'PmT#fA#mT.mT.mA.mA.mC.mC.mA.mA.mG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 37); 5' PmT#fC#mT.mT.mTJC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 38);

[0048] 5' PmT#fA#mT.mC.mA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 39);

[0049] 5'PmT#fA#mT.mT.mA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 40);

[0050] 5PmT#fC#mT.mT.mT.mC.mT.mT.mT.fG.fG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 41);

[0051] 5' PmT#fA#mT.mC.mA.mG.mT.mC.mT.fT.fr.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 42);

[0052] 5' PmT#fA#mT.mT.mA.mA.mC.mC.mA.fA.fG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0053] 43);

[0054] 5' PmT#fC#mTK.fT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#niC 3' (SEQ ID NO:

[0055] 44);

[0056] 5' PmT#fA#mT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fr#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:45);

[0057] 5' PmT#fA#mT.1T.fA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0058] 46);

[0059] 5' PmT#fC#r.fT.1T.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:

[0060] 47);

[0061] 5' PmT#fA#fT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:

[0062] 48);

[0063] 5' PmT#fA#fr.fT.fA.fA.mC.mC.mA.mA.mG.mA.mA.fr#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0064] 49);

[0065] 5' PmT#fC#mT.fT.fT.fC.fT.fr.mT.fG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 31(SEQ ID NO: 50);

[0066] 5' PmT#fA#mT.fC.fA.fG.fT.fC.mT.fr.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 51);

[0067] 5' PmT#fA#mT.1T.fA.fA.fC.fC.mA.fA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 52);

[0068] 5' PmT#fC#mT.fT.mT.fC.mT.fr.mT.fG.mG.fr.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 53);

[0069] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.1T.mT.fC.mC.1T#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:54); 5' PmT#fA#mT.fT.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 55);

[0070] 5' PmT#fC#mT.fT.mT.fC.mT.fT.mT.fG.mG.fT.mC.fT#mG#fC#mA#fT#mT#fC 3' (SEQ ID NO: 56);

[0071] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fT.mT.fC.mC.fT#mG#fG#mT#fG#mA#fG 3' (SEQ ID NO: 57);

[0072] 5' PmT#fA#mT.fr.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#fC#mT#fG#mA#fA 3' (SEQ ID NO: 58); wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0073] In embodiments, disclosed herein is a pharmaceutical composition comprising the composition described herein and a pharmaceutically acceptable carrier. In embodiments, the pharmaceutical composition is suitable for self-delivery.

[0074] In embodiments, the pharmaceutical composition further comprises a vehicle. In embodiments, the vehicle is one or more of calcium phosphate, a cationic lipid, a cationic polymer, polyethyleneimine and a proteinbased transfection reagent.

[0075] In embodiments, the composition is formulated for topical, pulmonary, or parenteral delivery.

[0076] In embodiments, disclosed herein is a cell comprising the composition or pharmaceutical composition described in any of the embodiments disclosed herein.

[0077] In aspects, disclosed herein is a method of promoting or enhancing degradation of an mRNA encoding USP10, comprising: administering to a subject in need thereof an effective amount of the composition of any one of the embodiments disclosed herein, or pharmaceutical composition of any one of the embodiments disclosed herein, or contacting a cell with the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, and administering to a subject in need thereof an effective amount of the composition, wherein the promoting or enhancing is as compared to a state before or without composition.

[0078] In embodiments, the method is performed in vitro. In embodiments, the method is performed ex vivo.

[0079] In embodiments, the method is performed in vivo.

[0080] In aspects, disclosed herein is a method of treating a disease or disorder, comprising: administering to a subject in need thereof an effective amount of the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, or contacting a cell with the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein and administering to a subject in need thereof an effective amount of the composition.

[0081] In embodiments, the disease or disorder is characterized by scarring.

[0082] In embodiments, the disease or disorder is selected from ocular scarring (optionally selected from scarring of the cornea and / or retina), integumentary scarring, internal organ scarring, and internal organ (optionally selected from lung, liver, and kidney) fibrosis.

[0083] In aspects, disclosed herein is a method of reducing or eliminating ocular scarring in an eye of a subject after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0084] In aspects, disclosed herein is a method for accelerating wound closure in an eye of a subject after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the accelerating is compared to upregulation without treatment or pre-treatment.

[0085] In aspects, disclosed herein is a method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of a subject after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the suppressing is compared to upregulation without treatment or pre-treatment.

[0086] In aspects, disclosed herein is a method of eliminating or reducing fibrosis of a subject after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment.

[0087] In aspects, disclosed herein is a method of eliminating or reducing scarring in a skin of a subject after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment.

[0088] In aspects, disclosed herein is a method of eliminating or reducing scarring in an eye of a subject as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0089] In embodiments, the composition is administered at a frequency of no more than about once monthly, no more than about once every two months.

[0090] In embodiments, the method further comprises administering one or more of an antibiotic and a steroid agent.

[0091] In embodiments, the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0092] In embodiments, the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment.

[0093] In embodiments, the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0094] In embodiments, the composition of any one of the embodiments disclosed herein, or the pharmaceutical composition of any one of the embodiments disclosed herein, or the cell of any one of the embodiments disclosed herein, or the method of any one of the embodiments disclosed herein, wherein the USP10 mRNA or protein is from a mammal.

[0095] In embodiments, the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0096] In aspects, disclosed herein is a method of reducing or eliminating ocular scarring in an eye of an animal after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human. In aspects, disclosed herein is a method for accelerating wound closure in an eye of an animal after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the accelerating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0097] In aspects, disclosed herein is a method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of an animal after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the suppressing is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0098] In aspects, disclosed herein is a method of eliminating or reducing fibrosis of an animal after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0099] In aspects, disclosed herein is a method of eliminating or reducing scarring in a skin of an animal after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0100] In aspects, disclosed herein is a method of eliminating or reducing scarring in an eye of an animal as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0101] In embodiments, the composition is administered at a frequency of no more than about once monthly, no more than about once every two months. In embodiments, the method further comprises administering one or more of an antibiotic and a steroid agent. In embodiments, the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0102] In embodiments, the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment. In embodiments, the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0103] BRIEF DESCRIPTION OF THE DRAWINGS

[0104] FIG. 1A and FIG. 1 B show the results of a reporter screening assay of USP10 sdRNA compounds. The results are shown graphically (FIG. 1A) and statistically (FIG. 1B). For each sdRNA compound listed on the x-axis of FIG. 1A, the left bar is 1 pM and the right bar is 0.1 pM. In FIG. 1B, the black arrow points to sdRNA compounds that performed as well or better than the “original.” In FIG. 1 A, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 / SEQ ID NO: 2); (2) s17_ori / as_2 (SEQ ID NO: 8 / SEQ ID NO: 15); (3) s17_ori I as_3 (SEQ ID NO: 8 1 SEQ ID NO: 16); (4) s17_ori / as_4 (SEQ ID NO: 8 1 SEQ ID NO: 17); (5) s17_ori I as_5 (SEQ ID NO: 8 / SEQ ID NO: 18); (6) s17_ori / as_P3 (SEQ ID NO: 8 / SEQ ID NO: 19; (7) s17_ori I as_P5 (SEQ ID NO: 8 1 SEQ ID NO: 20); (8) s17_allM I as_ori (SEQ ID NO: 12 / SEQ ID NO: 2); (9) s17_allM / as_2 (SEQ ID NO: 12 / SEQ ID NO: 15); (10) s17_allM I as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (11 ) s17_all M / as_4 (SEQ I D NO: 12 / SEQ ID NO: 17); (12) s17_all M / as_5 (SEQ ID NO: 12 / SEQ I D NO: 18); (13) s17_allM / as_P3 (SEQ ID NO: 12 / SEQ I D NO: 19); (14) s17_allM / as_P5 (SEQ ID NO: 12 / SEQ ID NO: 20); (15) s17_P3 / as_ori (SEQ ID NO: 13 / SEQ ID NO: 2); (16) s17_P3 / as_2 (SEQ ID NO: 13 / SEQ ID NO: 15); (17) s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (18) s17_P3 / as_4 (SEQ ID NO: 13 / SEQ ID NO: 17); (19) s17_P31 as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (20) s17_P3 / as_P3 (SEQ ID NO: 13 / SEQ ID NO: 19); (21) s17_P3 / as_P5 (SEQ ID NO: 13 / SEQ ID NO: 20); (22) s17_P5 1 as_ori (SEQ ID NO: 14 / SEQ ID NO: 2); (23) s17_P5 / as_2 (SEQ ID NO: 14 / SEQ ID NO: 15); (24) s17_P51 as_3 (SEQ ID NO: 14 1 SEQ ID NO: 16); (25) s17_P5 / as_4 (SEQ ID NO: 14 / SEQ ID NO: 17); (26) s17_P5 I as_5 (SEQ ID NO: 141 SEQ ID NO: 18); (27) s17_P5 1 as_P3 (SEQ ID NO: 14 / SEQ ID NO: 19); and (28) s17_P5 1 as_P5 (SEQ ID NO: 14 / SEQ ID NO: 20). In FIG. 1A, “VO” stands for “vector only.”

[0105] In FIG. 1B, the references in each column, starting from the left side of FIG. 1 B, refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 / SEQ ID NO: 2); (2) s17_ori / as_2 (SEQ ID NO: 8 / SEQ ID NO: 15); (3) s17_ori / as_3 (SEQ ID NO: 8 1 SEQ ID NO: 16); (4) s17_ori / as_4 (SEQ ID NO: 8 / SEQ ID NO: 17); (5) s17_ori / as_5 (SEQ ID NO: 8 1 SEQ ID NO: 18); (6) s17_ori I as_P3 (SEQ ID NO: 8 1 SEQ ID NO: 19; (7) s17_ori I as_P5 (SEQ ID NO: 8 / SEQ ID NO: 20); (8) s17_allM / as_ori (SEQ ID NO: 12 / SEQ ID NO: 2); (9) s17_allM / as_2 (SEQ ID NO: 12 / SEQ ID NO: 15); (10) s17_allM / as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (11) s17_allM / as_4 (SEQ ID NO: 12 / SEQ ID NO: 17); (12) s17_allM / as_5 (SEQ ID NO: 12 / SEQ ID NO: 18); (13) s17_allM / as_P3 (SEQ ID NO: 12 / SEQ ID NO: 19); (14) s17_allM I as_P5 (SEQ ID NO: 12 / SEQ ID NO: 20); (15) s17_P3 / as_ori (SEQ ID NO: 13 / SEQ ID NO: 2); (16) s17_P3 / as_2 (SEQ ID NO: 13 / SEQ ID NO: 15); (17) s17_P3 I as_3 (SEQ I D NO: 13 / SEQ I D NO: 16); (18) s17_P3 / as_4 (SEQ I D NO: 13 / SEQ I D NO: 17); (19) s17_P3 I as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (20) s17_P3 / as_P3 (SEQ ID NO: 13 / SEQ ID NO: 19); (21) s17_P3 1 as_P5 (SEQ ID NO: 13 / SEQ ID NO: 20); (22) s17_P5 / as_ori (SEQ ID NO: 14 / SEQ ID NO: 2); (23) s17_P5 1 as_2 (SEQ ID NO: 14 / SEQ ID NO: 15); (24) s17_P5 1 as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (25) s17_P5 / as_4 (SEQ ID NO: 14 / SEQ ID NO: 17); (26) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (27) s17_P5 / as_P3 (SEQ ID NO: 14 / SEQ ID NO: 19); and (28) s17_P5 / as_P5 (SEQ ID NO: 14 / SEQ ID NO: 20).

[0106] FIG. 2A, FIG. 2B, and FIG. 2C show the results of reporter dose curves of USP10 sdRNA compounds. The results are shown graphically (FIG. 2A) and statistically (FIG. 2B). In FIG. 2A, the references across the X- axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 / SEQ ID NO: 2); (2) s17_ori / as_3 (SEQ ID NO: 8 / SEQ ID NO: 16); (3) s17_al IM / as_3 (SEQ I D NO: 12 / SEQ I D NO: 16); (4) s17_P 31 as_3 (SEQ I D NO: 13 / SEQ I D NO: 16); (5) s17_P5 / as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (6) s17_ori / as_P5 (SEQ ID NO: 8 / SEQ ID NO: 20); (7) s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (8) s17_P5 1 as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (9) US36_s_15 1 US36_as (SEQ ID NO: 1 1 SEQ ID NO: 2); and (10) US36_s_17 / US36_as (SEQ ID NO: 8 / SEQ ID NO: 2). In FIG. 2A, “NTC” stands for “Non-Template Control” (NTC_s / NTC.as (SEQ ID NO: 21 I SEQ ID NO: 22). In FIG. 2B, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 I SEQ ID NO: 2); (2) s17_ori / as_3 (SEQ ID NO: 8 / SEQ ID NO: 16); (3) s17_allM I as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (4) s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (5) s17_P5 1 as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (6) s17_ori / as_P5 (SEQ ID NO: 8 1 SEQ ID NO: 20); (7) s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (8) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (9) US36_s_15 / US36_as (SEQ ID NO: 1 I SEQ ID NO: 2); and (10) US36_s_17 1 US36_as (SEQ ID NO: 8 / SEQ ID NO: 2). In FIG. 2C, the references across the top of FIG. 2C ( / .e., sense strand sequences) are the following: (1) s17_ori (SEQ ID NO: 8);(2) s17_allM (SEQ ID NO: 12); (3) s17_P3 (SEQ ID NO: 13); and (4) s17_P5 (SEQ ID NO: 14). In FIG. 2C, the references across the left side of FIG. 2C ( / .e., anti-sense strand sequences) are the following: (1) as_ori (SEQ ID NO: 2); (2) as_2 (SEQ ID NO: 15); (3) as_3 (SEQ ID NO: 16); (4) as_4 (SEQ ID NO: 17); (5) as_5 (SEQ ID NO: 18); (6) as_P3 (SEQ ID NO: 19) and (7) as_P5 (SEQ ID NO: 20). For each compound listed on the x-axis of FIG. 2A, the order of bars on the graph is as follows from left to right: 1 pM, 0.2 pM, 0.04 pM, 0.008 pM, 0.0016 pM, 0.00032 pM, and 0.000064 pM. FIG. 2C shows the best candidates from the screening assay in green (e.g., see circles); ( / .e., s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); and s17_P5 1 as_5 (SEQ ID NO: 14 / SEQ ID NO: 18).

[0107] FIG. 3A and FIG. 3B show the results of a reporter screening assay of USP10 sdRNA compounds. The results are shown graphically (FIG. 3A) and statistically (FIG. 3B). In FIG. 3A, the references across the X- axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_12 1 US36_as (SEQ ID NO: 4 / SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 / SEQ ID NO: 2); (3) US36_s_141 US36_as (SEQ ID NO: 61 SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 I SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 I SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 1 SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 I SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36 old”) (SEQ ID NO: 1 1 SEQ ID NO: 2). In FIG. 3A, “NTC” stands for “Non-Template Control” (NTC_s / NTC.as (SEQ ID NO: 21 / SEQ ID NO: 22). For each sdRNA compound listed on the x-axis of FIG. 3A, the order of bars on the graph is as follows from left to right: 1 pM, 0.2 pM, 0.04 pM, 0.008 pM, 0.0016 pM, 0.00032 pM, and 0.000064 pM. In FIG. 3B, the references across the top of FIG. 3B refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_121 US36_as (SEQ ID NO: 4 / SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 I SEQ ID NO: 2); (3) US36_s_14 / US36_as (SEQ ID NO: 6 / SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 I SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 1 SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 1 SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36 old”) (SEQ ID NO: 1 / SEQ ID NO: 2).

[0108] FIG. 4 is a graph showing the results of a reporter screening assay of USP10 sdRNA compounds. For each sdRNA compound listed on the x-axis of FIG. 4, the left bar is 1 pM and the right bar is 0.1 pM. In FIG. 4, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_12 I US36_as (SEQ ID NO: 4 I SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 / SEQ ID NO: 2); (3) US36_s_14 / US36_as (SEQ ID NO: 6 1 SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 I SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 1 SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 / SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36” is same as“US36 old” in FIG. 3A and FIG. 3B) (SEQ ID NO: 1 1 SEQ ID NO: 2). In FIG. 4, “NTC” stands for “Non-Template Control” (NTC-S I NTC.as (SEQ ID NO: 21 / SEQ ID NO: 22), GAPDH is the following: GAPDH.s I GAPDH.as (SEQ ID NO: 251 SEQ ID NO: 26); and MAP4K4 is the following: MAP4K4_s I MAP4K4_as (SEQ ID NO: 23 / SEQ ID NO: 24).

[0109] FIG. 5A and FIG. 5B show the results of qPCR analysis performed in primary human corneal fibroblast cells. The results are shown graphically (FIG. 5A) and statistically (FIG. 5B). In FIG. 5A, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_15_Alt (SEQ ID NO: 1 / SEQ ID NO: 2); (2) US36_17_Alt (SEQ ID NO: 8 / SEQ ID NO: 2); (3) US36_1 (SEQ ID NO: 8 / SEQ ID NO: 16); (4) US36_2 (SEQ ID NO: 14 / SEQ ID NO: 16); (5) US36_3 (SEQ ID NO: 8 / SEQ ID NO: 18); and (6) US36.4 (SEQ ID NO: 12 / SEQ ID NO: 16). In FIG. 5A, “NTC” stands for “Non-Template Control” (NTC_s / NTC.as (SEQ ID NO: 21 / SEQ ID NO: 22). For each sdRNA compound listed on the x-axis of FIG. 5A, the order of bars on the graph is as follows from left to right: 2.0 pM, 1.0 pM, 0.5 pM, 0.25 pM, 0.125 pM, 0.0625 pM, and 0.0313 pM. In FIG. 5B, the references in the first column on the left refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): 1) US36_15_Alt (SEQ ID NO: 1 1 SEQ ID NO: 2); (2) US36_17_Alt (SEQ ID NO: 8 / SEQ ID NO: 2); (3) US36_1 (SEQ ID NO: 8 / SEQ ID NO: 16); (4) US36_2 (SEQ ID NO: 14 / SEQ ID NO: 16); (5) US36_3 (SEQ ID NO: 8 1 SEQ ID NO: 18); and (6) US36_4 (SEQ ID NO: 12 / SEQ ID NO: 16).

[0110] FIG. 6 is a non-limiting image showing an in vivo corneal scrape. FIG. 7A, FIG. 7B, and FIG. 7C are graphs and images showing how US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) prevent wound re-opening on Day 28 in scrape PCED mouse model in male mice. FIG. 7A shows a non-limiting schematic workflow of experiments in scrape- induced PCED mouse model. FIG. 7B shows representative fluorescein-stained corneas and demonstrate that US16 (SEQ ID NO: 57 I SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatments effectively prevent epithelial wound re-opening at 4 weeks post-PCED epithelial scrape. FIG. 7C shows quantification of fluorescein-stained corneas using FIJI demonstrates a significant reduction in the open wound area in the US16 (SEQ ID NO: 57 / SEQ ID NO: 58) treatment group compared to the controls (PBS and Non-template control (“NTC”; (SEQ ID NO: 69 / SEQ ID NO: 70)) N=14 eyes / condition.

[0111] FIG. 8A, FIG. 8B, FIG. 8C, FIG. 8D, FIG. 8E, and FIG. 8F are graphs and images showing how US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) reduce corneal scarring in male mice. FIG. 8A shows optical coherence tomography (OCT) imaging reveals the preservation of corneal structure with sdRNA treatment. FIG. 8B and FIG. 8C show how quantification of central corneal thickness measured with FIJI confirms restoration to near-normal levels in drug-treated eyes (N=14 eyes / PBS and US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60); N=6 eyes / NTC (SEQ ID NO: 69 / SEQ ID NO: 70) and Unwounded. FIG. 8D shows central corneal thickness. FIG. 8E shows how tonometry measurements indicate no significant changes in intraocular pressure (IOP) among treatment and control groups at Day 28, however there is a trend towards lower IOP with US16 (SEQ ID NO: 571 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60). N=8 eyes / condition. FIG. 8F shows normalized fibrotic gene expression analysis by RT-qPCR demonstrates decreasing trend in US16 (SEQ ID NO: 57 I SEQ ID NO: 58) -treated group compared to PBS control, which is highly variable N=8 eyes / condition; 2 eyes pooled per sample for RNA isolation.

[0112] FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, and FIG. 9E are graphs and images showing how US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) reduce corneal scarring, in female mice. In FIG. 9A, similar to male mice, the corneal epithelium at Day 28, exhibited recurrent wound re-opening and corneal abrasions in control groups (PBS and NTC (SEQ ID NO: 69 / SEQ ID NO: 70)) while the drug-treated groups (US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) did not reopen. In total, 9 / 10 corneas in the drug-treated groups demonstrated no reopening compared to only 1 / 10 in the control groups. FIG. 9B shows the average normalized open wound area was significantly reduced in the US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60)-treated corneas by 91.71 % (p<0.001) and 97.26% (p<0.0001), respectively, compared to PBS-treated corneas. FIG. 9C shows OCT imaging at Day 28 showed similar corneal morphology in drug-treated female mice as seen in the male mice, with intact epithelium and endothelium and improved morphology with sdRN A treatment. FIG. 9D and FIG. 9E show pixel intensity analysis further corroborated these findings, showing a 30.8% (p<0.0001) reduction in scarring in the US16 (SEQ ID NO: 57 1 SEQ ID NO: 58)-treated group and a 31.77% reduction (p<0.0001 ) in the US16.1 -treated group compared to PBS but did not reach the pixel intensity of unwounded. Both controls, PBS and NTC (SEQ ID NO: 69 1 SEQ ID NO: 70) showed comparable mean pixel intensities (p=0.1810) and were not significantly different. N=6 eyes / condition.

[0113] FIG. 10A and FIG. 10B is a graph and image showing corneal cell morphology and endothelial cell density is not affected by US16 (SEQ ID NO: 57 / SEQ ID NO: 58) or US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatment: US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) are nontoxic in male mice. FIG. 10A shows in vivo confocal images that have no differences in endothelial cell size, shape, or morphology across treated and control groups. Stromal keratocytes maintain their usual organized patterns in all groups. FIG. 10B is a graph of endothelial cell density analysis on HEYEX software reveals no significant changes in US16 (SEQ ID NO: 571 SEQ ID NO: 58)-treated groups compared to PBS control and unwounded N=4 eyes / condition.

[0114] FIG. 11 A, FIG. 11 B, and FIG. 11C are H&E images after wounding and treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) in male mice. FIG. 11 A shows an H&E image for NTC (SEQ ID NO: 69 / SEQ ID NO: 70), FIG. 11 B shows an H&E image for US16 (SEQ ID NO: 57 / SEQ ID NO: 58), and FIG. 11 C shows an H&E image for US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60).

[0115] FIG. 12A, FIG. 12B, and FIG. 12C are H&E images after wounding and treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) in female mice. FIG. 12A shows an H&E image for PBS, FIG. 12B shows an H&E image for US16 (SEQ ID NO: 57 / SEQ ID NO: 58), and FIG. 12C shows an H&E image for US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60).

[0116] FIG. 13 shows immunohistochemistry images of hemidesmosomes after wounding and treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) in male mice.

[0117] FIG. 14A, FIG. 14B, FIG. 14C, FIG. 14D, and FIG. 14E are graphs and images showing the safety of USP10 knockdown with USP10-targeting siRNA (US09 (SEQ ID NO: 61 1 SEQ ID NO: 62)) after epithelial scratch in female and male New Zealand rabbits were wounded with a light scratch wound to the central cornea without disruption of the basement membrane. FIG. 14A shows a graph of central corneal thickness. FIG. 14B shows a graph of IOP. FIG. 14C shows a graph of schirmer tear strips, and FIG. 14D shows a graph when weight was not significantly different. FIG. 14E shows images of eye health showed no signs of ocular inflammation, corneal haze, neovascularization, chemosis, or other ocular anomaly of the cornea, conjunctiva, or sclera after sdRNA treatment.

[0118] FIG. 15A, FIG. 15B, and FIG. 15C are images showing the safety of USP10 knockdown with USP10-targeting siRNA (US09 (SEQ ID NO: 61 / SEQ ID NO: 62)) after epithelial scratch continued in female and male New Zealand Rabbits. FIG. 15A is an image of how OCT showed no differences in the treated groups compared to unwounded rabbit corneas. FIG. 15B shows representative confocal images (HRT3), there was no noticeable variation in the morphology in each layer of rabbit cornea. Stromal layers showed an organized keratocyte pattern with basal nerve plexuses. Examples shown are female rabbits. No sex dependent differences were observed. FIG. 15C are images showing the endothelial layer of all groups up to 3.5 months demonstrated normal hexagonal pattern with no toxicity with US09 (SEQ ID NO: 61 I SEQ ID NO: 62) or US09.1 (SEQ ID NO: 63 1 SEQ ID NO: 64).

[0119] FIG. 16 is an image showing USP10 knockdown with US36.1 (SEQ ID NO: 67 / SEQ ID NO: 68) in ex vivo human corneas.

[0120] DETAILED DESCRIPTION

[0121] The present disclosure demonstrates, inter alia, that incorporation of at least one modification in at least one position of a sense strand of a self-deliverable RNA (sdRNA), or altering the length of a sense strand of a sdRNA, inhibits the expression of a ubiquitin specific peptidase 10 (USP10) gene. The use of these modifications and / or sense strand length alterations, e.g. in the context of a chemically modified sdRNA (e.g., one or more of a locked nucleic acid (LNA), a phosphorothioate, and a 2'-O-Methyl modifications), is suitable for causing a reduction of expression and / or activity of a USP10 mRNA or protein in vitro and in vivo, and is useful in treating a variety of diseases and disorders.

[0122] In aspects, disclosed herein is a composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, and the sense strand is about 16 to about 18 nucleotides in length and has sequence complementarity to the antisense strand. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. In embodiments, the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10. In embodiments, the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

[0123] In embodiments, the sense strand is about 16 nucleotides in length, or is about 17 nucleotides in length, or is about 18 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 16 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 17 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, the nucleic acid complex has: a sense strand consisting of 18 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length. In embodiments, at least 16, 17, 18, 19, or 20 nucleotides of the antisense strand are complementary to the target.

[0124] In embodiments, the nucleic acid complex is an interfering nucleic acids and generally includes a sequence of cyclic subunits, each bearing a base-pairing moiety, linked by intersubunit linkages that allow the basepairing moieties to hybridize to a target sequence in a nucleic acid (typically RNA) by Watson-Crick base pairing, to form a nucleic acid: oligomer heteroduplex within the target sequence. Interfering RNA molecules include, but are not limited to, antisense molecules, siRNA molecules, single-stranded siRNA molecules, miRNA molecules and shRNA molecules. Such an interfering nucleic acids can be designed to block or inhibit translation of mRNA or to inhibit natural pre-mRNA splice processing, or induce degradation of targeted mRNAs, and may be said to be directed to or targeted against a target sequence with which it hybridizes. Interfering nucleic acids may include, for example, peptide nucleic acids (PNAs), locked nucleic acids (LNAs), 2'-fluoro, 2'-O-Methyl oligonucleotides and RNA interference agents (siRNA agents). RNAi molecules generally act by forming a heteroduplex with the target molecule, which is selectively degraded or knocked down, hence inactivating the target RNA. Under some conditions, an interfering RNA molecule can also inactivate a target transcript by repressing transcript translation and / or inhibiting transcription of the transcript. An interfering nucleic acid is more generally said to be targeted against a biologically relevant target, such as a protein, when it is targeted against the nucleic acid of the target in the manner described above.

[0125] In embodiments, the nucleic acid complex specifically hybridizes to a target polynucleotide if the oligonucleotide hybridizes to the target under physiological conditions, with a Tm substantially greater than about 45°C, or at least about 50°C, or at least about 60°C to about 80°C or higher. Such hybridization corresponds to stringent hybridization conditions. At a given ionic strength and pH, the Tm is the temperature at which about 50% of a target sequence hybridizes to a complementary polynucleotide. Such hybridization may occur with near or substantial complementarity of the antisense oligomer to the target sequence, as well as with exact complementarity.

[0126] In embodiments, the target is or comprises mRNA (messenger RNA), microRNA, piRNA (piwi-interacting RNA), a coding DNA sequence or a noncoding DNA sequence.

[0127] In embodiments, the target is or comprises mammalian mRNA or viral mRNA. In embodiments, the target is an intronic region of the mRNA.

[0128] In embodiments, the target is ubiquitin specific peptidase (USP10), an enzyme encoded by the USP10 gene. The gene encodes a cysteine protease, an enzyme that specifically cleaves ubiquitin-conjugated protein substrates. Further, the protein is a deubiquitinase that can remove conjugated ubiquitin from target proteins such as p53 / TP53, BECN1 , SNX3 and CFTR. In response to DNA damage, USP10 is translocated to the nucleus where it is involved in the deubiquitination of p53. In embodiments, the USP10 mRNA or protein is from a mammal. In embodiments, the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit guinea pig, and human.

[0129] The following table shows a set of the nucleic acid complex sequences disclosed herein:

[0130] Table 1 : Unmodified siRNA and Target Site Sequences

[0131] In embodiments, the sense strand comprises the base sequence of SEQ ID NOs : 1 -2, or a variant thereof. In embodiments, the antisense strand comprises the base sequence of SEQ ID NO: 3 or a variant thereof. In embodiments, the variant comprises about 1, or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0132] In embodiments, the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 6 , or a variant thereof. In embodiments, the antisense strand consists of the base sequence of SEQ ID NO: 7 , or a variant thereof.

[0133] In embodiments, the nucleic acid complex disclosed herein can employ a variety of oligonucleotide chemistries. Examples of oligonucleotide chemistries include, without limitation, peptide nucleic acid (PNA), linked nucleic acid (LNA), phosphorothioate, 2'0-Me-modified oligonucleotides, and morpholino chemistries, including combinations of any of the foregoing. Phosphorothioate and 2'-0-Me- and 2'-F modified chemistries are often combined to generate 2'-O-Me / 2'-F-modified oligonucleotides having a phosphorothioate backbone. See, e.g., PCT Publication Nos. WO / 2021 / 174171 , WQ / 2013 / 112053 and WQ / 2009 / 008725, each of which is hereby incorporated by reference in its entirety.

[0134] In embodiments, the nucleic acid complex comprises at least one further modification selected from locked nucleic acid (LNA), phosphorothioate, 2'-O-Methyl, 2'-O-Methoxyethyl, 2'-O-alkyl-RNA unit, 2'-OMe-RNA unit, 2'-amino-DNA unit, 2'-fluoro-DNA unit, peptide nucleic acid (PNA) unit, hexitol nucleic acids (HNA) unit, INA unit, and 2'-O-(2-Methoxyethyl)-RNA (2' MOE RNA) unit modifications.

[0135] In embodiments, the phosphate backbone of at least one nucleotide in the oligonucleotide is substituted with a phosphorothioate linker (Ps).

[0136] In embodiments, the oligonucleotide comprises at least one of each locked nucleic acid (LNA), phosphorothioate, 2'-O-Methyl, 2'-O-Methoxyethyl, 2'-O-alkyl-RNA unit, 2'-OMe-RNA unit, 2 -amino-DNA unit, 2'-fluoro-DNA unit, peptide nucleic acid (PNA) unit, hexitol nucleic acids (HNA) unit, INA unit, and 2'-O- (2-Methoxyethyl)-RNA (2' MOE RNA) unit modifications.

[0137] 2'-O-methylated nucleosides carry a methyl group at the 2'-OH residue of the ribose molecule. 2'-O-Me-RNAs show the same (or similar) behavior as RNA, but are protected against nuclease degradation. 2'-O-Me-RNAs can also be combined with phosphothioate oligonucleotides (PTOs) for further stabilization. 2'-0-Me-RNAs (phosphodiester or phosphothioate) can be synthesized according to routine techniques in the art (see, e.g., Yoo et al., Nucleic Acids Res. 32:2008-16, 2004, which is hereby incorporated by reference).

[0138] In embodiments, the 2'-O-methyl nucleoside is positioned at or near the 3' terminus of the sense strand. In some embodiments, 3' terminal region of the sense strand comprises a plurality of 2'-O-methylated nucleosides (e.g., 2, 3, 4, 5 or 6 2'-O-methylated nucleosides within 6 nucleosides of the 3' terminus). In some embodiments, the 2'-O-methyl nucleoside is positioned at or near the 3' terminus of the antisense strand. In some embodiments, 3' terminal region of the antisense strand comprises a plurality of 2'-O- methylated nucleosides (e.g., 2, 3, 4, 5 or 6 2'-O-methylated nucleosides within 6 nucleosides of the 3' terminus). In some embodiments, both the 3' terminal region of the sense strand and the 3' terminal region of the antisense strand comprise a plurality of 2'-O-methylated nucleosides. In some embodiments, the sense strand comprises 2'-O-methylated nucleosides that alternate with unmodified nucleosides. In some embodiments, the sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 2 -O-methylated nucleosides that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises 2'-O-methylated nucleosides that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 2'-O-methylated nucleosides that alternate with unmodified nucleosides.

[0139] In embodiments, the 2'-F nucleoside (f) is positioned at the 3' terminus of the sense strand. In some embodiments, 3' terminal region of the sense strand comprises a plurality of 2'-F nucleosides (e.g., 2, 3, 4, 5 or 6 2'-F nucleosides within 6 nucleosides of the 3' terminus). In some embodiments, the 2'-F nucleoside is positioned at the 3' terminus of the antisense strand. In some embodiments, 3' terminal region of the antisense strand comprises a plurality of 2'-F nucleosides (e.g., 2, 3, 4, 5 or 6 2'-F nucleosides within 6 nucleosides of the 3' terminus). In some embodiments, both the 3' terminal region of the sense strand and the 3' terminal region of the antisense strand comprise a plurality of 2'-F nucleosides. In some embodiments, the sense strand comprises 2'-F nucleosides that alternate with unmodified nucleosides. In some embodiments, the sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 2'-F nucleosides that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises 2'-F nucleosides that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 2'-F nucleosides that alternate with unmodified nucleosides. In embodiments, the phosphorothioate linker (ps) is positioned at the 3' terminus of the sense strand. In some embodiments, 3' terminal region of the sense strand comprises a plurality of phosphorothioate linkers (e.g., 2, 3, 4, 5 or 6 phosphorothioate linkers within 6 nucleosides of the 3' terminus). In some embodiments, the phosphorothioate linker is positioned at the 3' terminus of the antisense strand. In some embodiments, 3' terminal region of the antisense strand comprises a plurality of phosphorothioate linkers (e.g., 2, 3, 4, 5 or 6 phosphorothioate linker within 6 nucleosides of the 3' terminus). In some embodiments, both the 3' terminal region of the sense strand and the 3' terminal region of the antisense strand comprise a plurality of phosphorothioate linkers. In some embodiments, the sense strand comprises phosphorothioate linkers that alternate with unmodified nucleosides. In some embodiments, the sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 phosphorothioate linkers that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises phosphorothioate linkers that alternate with unmodified nucleosides. In some embodiments, the anti-sense strand comprises a contiguous sequence of 2, 3, 4, 5, 6, 7 or 8 phosphorothioate linker that alternate with unmodified nucleosides.

[0140] In embodiments, the nucleic acid complex comprises one or more chemical modifications. In embodiments, the nucleic acid complex comprises one or more chemical modifications selected from Table 2.

[0141] Table 2 shows a set of the modified nucleic acid complex sequences disclosed herein:

[0142] Attorney Docket No. 137434-5001 -PC

[0143] SUNY RF Ref.: 110-2275

[0144] Table 2: Modified siRNA Sequences f represents 2’ Fluoro m represents 2’ O-methyl

[0145] Ps represents phosphorothioate linker

[0146] 5Phos represents 5’ phosphate

[0147] 3’CholTEG represents Cholesterol conjugate vP-mll represents 5'-Vinylphosphonate 2'0-mll

[0148] In embodiments, the chemical modification is a 2'-O-methylated nucleoside, a phosphorothioate bond, or a hydrophobic moiety. In embodiments, the chemical modification is selected from a locked nucleic acid (LNA), phosphorothioate, 2'-O-Methyl, 2'-O-Methoxyethyl, 2'-O-alkyl-RNA unit, 2'-0Me-RNA unit, 2 -amino-DNA unit, 2'-fluoro-DNA unit, peptide nucleic acid (PNA) unit, hexitol nucleic acids (HNA) unit, INA unit, and a 2'- O-(2-Methoxyethyl)-RNA (2' MOE RNA) unit. In embodiments, the chemical modification is a 2' fluoro group. In embodiments, the chemical modification is 2'-O-methyl (2'0Me) group. In embodiments, the chemical modification is a phosphorothioate linker. In embodiments, the chemical modification is a 5'- vinylphosphonate 2'0-mU group. In embodiments, the chemical modification is a cholesterol conjugate, or the chemical modification comprises docosanoic acid (DCA).

[0149] In embodiments, the composition comprises a modified sequence selected from: (a) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 9) 5'

[0150] [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 10) 5'

[0151] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][P s][mC][Ps][fG] 3'; (b) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 11) 5' [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 12) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fll][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][Ps][mC][ Ps][fG] 3'; (c) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 13) 5' [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 14) 5'

[0152] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][P s][mC][Ps][fG] 3'; and (d) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 15) 5' [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fll][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 16) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fll][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][Ps][fC][Ps][mC][ Ps][fG] 3'; wherein f represents 2' Fluoro, m represents 2' O-methyl, Ps represents phosphorothioate linker, 5Phos represents 5' phosphate, 3'CholTEG represents Cholesterol conjugate, and vP-mll represents 5'- Vinylphosphonate 2'0-mU. In embodiments, disclosed herein is a composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, the sense strand is about 15 nucleotides in length and having sequence complementarity to the antisense strand; and the antisense strand and the sense strand comprise one or more chemical modifications, selected from P, #, f, 2’ Fluor, m, 2’0-methyl, Choi 3’, cholesterol and docosanoic acid (DCA), wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0153] In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition. In embodiments, the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. In embodiments, the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

[0154] In embodiments, the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10. In embodiments, the nucleic acid complex has: a sense strand consisting of 15 nucleotides in length and an antisense strand consisting of 20 nucleotides in length.

[0155] In embodiments, the sense strand comprises the base sequence of SEQ ID NO: 5 or a variant thereof. In embodiments, the antisense strand comprises the base sequence of SEQ ID NO: 3 , or a variant thereof. In embodiments, the variant comprises about 1, or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0156] In embodiments, the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof. In embodiments, the antisense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof.

[0157] In embodiments, the nucleic acid complex comprises one or more chemical modifications. In embodiments, the nucleic acid complex comprises one or more chemical modifications selected from Table 3.

[0158] Table 3 shows a set of the modified nucleic acid complex sequences disclosed herein: Table 3: Modified siRNA Sequences

[0159]

[0160] P represents 5’ phosphate

[0161] # represents a phosphorothioate linker

[0162] 5 f represents 2' Fluoro m represents 2' O-methyl

[0163] Choi 3' represents Cholesterol conjugate

[0164] In embodiments, the composition comprises a modified sequence selected from a nucleic acid complex comprising: a sense strand selected from

[0165] 5' mC#mA#mG.mA.mC.mC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 17);

[0166] 5' mC#mA#mG.mG.mA.mA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 18);

[0167] 5' mT#mA#mT.mT.mC.mT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 19);

[0168] 5' mC#mA#mG.mA.fC.fC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 20);

[0169] 5' mC#mA#mG.mG.fA.fA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 21);

[0170] 5' mT#mA#mT.mT.fC.fT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 22);

[0171] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 23);

[0172] 5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 24);

[0173] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 25);

[0174] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.fA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 26);

[0175] 5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.fT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 27);

[0176] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.fT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 28);

[0177] 5' mC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#mA-Chol 3' (SEQ ID NO: 29);

[0178] 5' mC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#mA-Chol 3' (SEQ ID NO: 30);

[0179] 5' mT#mA#fr.mT.fC.mT.fT.mG.fG.mT.fT.mA.fA#mT#mA-Chol 3' (SEQ ID NO: 31);

[0180] 5' fC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#fA-Chol 3' (SEQ ID NO: 32);

[0181] 5' fC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#fA-Chol 3' (SEQ ID NO: 33);

[0182] 5' fT#mA#fT.mT.fC.mT.1T.mG.fG.mT.1T.mA.fA#mT#fA-Chol 3' (SEQ ID NO: 34); and an antisense strand of:

[0183] 5' PmT#fC#mT.mT.mT.mC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 35);

[0184] 5' mT#fA#mT.mC.mA.mG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 36);

[0185] 5'PmT#fA#mT.mT.mA.mA.mC.mC.mA.mA.mG.mA.mA.fr#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 37);

[0186] 5' PmT#fC#mT.mT.mT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 38);

[0187] 5' PmT#fA#mT.mC.mA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 39); 5'PmT#fA#mT.mT.mA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 40);

[0188] 5'PmT#fC#mT.mT.mT.mC.mT.mT.mT.fG.fG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 41);

[0189] 5' PmT#fA#mT.mC.mA.mG.mT.mC.mT.fT.1T.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 42);

[0190] 5' PmT#fA#mT.mT.mA.mA.mC.mC.mA.fA.fG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0191] 43);

[0192] 5' PmT#fC#mT.fT.fT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:

[0193] 44);

[0194] 5' PmT#fA#mT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.1T#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:45);

[0195] 5' PmT#fA#mT.fr.fA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0196] 46);

[0197] 5' PmT#fC#IT.fT.fr.fC.mT.mT.mT.mG.mG.mT.mC.1T#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:

[0198] 47);

[0199] 5' PmT#fA#IT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:

[0200] 48);

[0201] 5' PmT#fA#fT.fr.fA.fA.mC.mC.mA.mA.mG.mA.mA.1T#niA#niC#mT#mG#mA#mA 3' (SEQ ID NO:

[0202] 49);

[0203] 5' PmT#fC#mT.fr.fT.fC.fr.fT.mT.fG.mG.mT.mC.fr#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 50);

[0204] 5' PmT#fA#mT.fC.fA.fG.fT.fC.mT.1T.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 51);

[0205] 5' PmT#fA#mT.1T.fA.fA.fC.fC.mA.fA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 52);

[0206] 5' PmT#fC#mTK.mTJC.mT.fr.mTJG.mG.fT.mC.r#mG#mC#mA#niT#mT#mC 3' (SEQ ID NO: 53);

[0207] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fr.mT.fC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:54);

[0208] 5' PmT#fA#mT.fT.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 55);

[0209] 5' PmT#fC#mT.fT.mT.fC.mT.1T.mT.fG.mG.fT.mC.fT#mG#fC#mA#fT#mT#fC 3' (SEQ ID NO: 56);

[0210] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fT.mT.fC.mC.fr#mG#fG#mT#fG#mA#fG 3' (SEQ ID NO: 57); 5' PmT#fA#mT.fr.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#fC#mT#fG#mA#fA 3' (SEQ ID NO: 58); wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0211] In embodiments, the nucleic acid complex comprises a phosphorothioate bond. Phosphorothioates (or S- oligos) are a variant of normal DNA in which one of the non-bridging oxygens is replaced by a sulfur. The sulfurization of the internucleotide bond reduces the action of endo-and exonucleases including 5' to 3' and 3' to 5' DNA POL 1 exonuclease, nucleases S1 and P1, RNases, serum nucleases and snake venom phosphodiesterase. Phosphorothioates are made by two principal routes: by the action of a solution of elemental sulfur in carbon disulfide on a hydrogen phosphonate, or by the method of sulfurizing phosphite triesters with either tetraethylthiuram disulfide (TETD) or 3H-1 , 2-ben zodithiol-3-one 1 , 1 -dioxide (BDTD) (see, e.g., Iyer et al., J. Org. Chem. 55, 4693-4699, 1990) or DDTT (Guzaev, Tetrahedron Letters 52, 434-437, 2011). The latter methods avoid the problem of elemental sulfur’s insolubility in most organic solvents and the toxicity of carbon disulfide. The TETD, BDTD, and DDTT methods also yield higher purity phosphorothioates.

[0212] In embodiments, disclosed herein is a pharmaceutical composition comprising the composition described herein and a pharmaceutically acceptable carrier. In embodiments, the pharmaceutical composition is suitable for self-delivery.

[0213] In embodiments, the pharmaceutical composition further comprises a vehicle. In embodiments, the vehicle is one or more of calcium phosphate, a cationic lipid, a cationic polymer, polyethyleneimine and a proteinbased transfection reagent.

[0214] In embodiments, the composition is formulated for topical, pulmonary, or parenteral delivery.

[0215] A pharmaceutically-acceptable carrier, in embodiments, is a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material. A pharmaceutically-acceptable carrier, in embodiments, may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate and mineral oils. In embodiments, the composition described herein is complexed with or associated with a lipid, the lipid optionally being selected from cholesterol, tocopherol, and a long-chain fatty acid having 10 or more carbon atoms.

[0216] In embodiments, the composition described herein is complexed with or associated with N- acetylgalactosamine (GalNac).

[0217] The pharmaceutical composition may additionally contain fillers, anti-aggregating agents, lubricants, wetting agents, perfumes, emulsifiers and preservatives. Also, the pharmaceutical composition of the present invention may be formulated using a method well known in the art, such that it can provide the rapid, sustained or delayed release of the active ingredient after administration to mammals. The formulation may be in the form of sterile injection solutions, etc.

[0218] In aspects, there is provided a method of treating or preventing a disease or disorder comprising administering to a subject in need thereof a therapeutically effective amount of the pharmaceutical composition described herein.

[0219] In embodiments, disclosed herein is a cell comprising the composition or pharmaceutical composition described in any of the embodiments disclosed herein.

[0220] In aspects, disclosed herein is a method of promoting or enhancing degradation of an mRNA encoding USP10, comprising: administering to a subject in need thereof an effective amount of the composition of any one of the embodiments disclosed herein, or pharmaceutical composition of any one of the embodiments disclosed herein, or contacting a cell with the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, and administering to a subject in need thereof an effective amount of the composition, wherein the promoting or enhancing is as compared to a state before or without composition.

[0221] In embodiments, the method is performed in vitro. In embodiments, the method is performed ex vivo.

[0222] In embodiments, the method is performed in vivo.

[0223] In aspects, disclosed herein is a method of treating a disease or disorder, comprising: administering to a subject in need thereof an effective amount of the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, or contacting a cell with the composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein and administering to a subject in need thereof an effective amount of the composition.

[0224] In embodiments, the disease or disorder is characterized by scarring.

[0225] In embodiments, the disease or disorder is selected from ocular scarring (optionally selected from scarring of the cornea and / or retina), integumentary scarring, internal organ scarring, and internal organ (optionally selected from lung, liver, and kidney) fibrosis.

[0226] In aspects, disclosed herein is a method of reducing or eliminating ocular scarring in an eye of a subject after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0227] In aspects, disclosed herein is a method for accelerating wound closure in an eye of a subject after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the accelerating is compared to upregulation without treatment or pre-treatment.

[0228] In aspects, disclosed herein is a method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of a subject after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the suppressing is compared to upregulation without treatment or pre-treatment.

[0229] In aspects, disclosed herein is a method of eliminating or reducing fibrosis of a subject after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment.

[0230] In aspects, disclosed herein is a method of eliminating or reducing scarring in a skin of a subject after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment.

[0231] In aspects, disclosed herein is a method of eliminating or reducing scarring in an eye of a subject as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0232] In embodiments, the composition is administered at a frequency of no more than about once monthly, no more than about once every two months.

[0233] In embodiments, the method further comprises administering one or more of an antibiotic and a steroid agent.

[0234] In embodiments, the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0235] In embodiments, the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment.

[0236] In embodiments, the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0237] In embodiments, the composition of any one of the embodiments disclosed herein, or the pharmaceutical composition of any one of the embodiments disclosed herein, or the cell of any one of the embodiments disclosed herein, or the method of any one of the embodiments disclosed herein, wherein the USP10 mRNA or protein is from a mammal.

[0238] In embodiments, the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0239] In aspects, disclosed herein is a method of reducing or eliminating ocular scarring in an eye of an animal after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human. In aspects, disclosed herein is a method for accelerating wound closure in an eye of an animal after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the accelerating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0240] In aspects, disclosed herein is a method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of an animal after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the suppressing is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0241] In aspects, disclosed herein is a method of eliminating or reducing fibrosis of an animal after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0242] In aspects, disclosed herein is a method of eliminating or reducing scarring in a skin of an animal after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pretreatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0243] In aspects, disclosed herein is a method of eliminating or reducing scarring in an eye of an animal as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of the embodiments disclosed herein or pharmaceutical composition of any one of the embodiments disclosed herein, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0244] In embodiments, the composition is administered at a frequency of no more than about once monthly, no more than about once every two months. In embodiments, the method further comprises administering one or more of an antibiotic and a steroid agent. In embodiments, the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0245] In embodiments, the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment. In embodiments, the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0246] This disclosure is further illustrated by the following non-limiting examples.

[0247] EXAMPLES

[0248] Example 1: Screening of USP 10 sdRNA Compounds with Alternative Modifications

[0249] In the experiments of this example, a dual dose screen of different sdRNA compound modifications was tested to determine the effect of potential patterns for the US36 target site. In these experiments, about 10,000 HeLa cells were seeded per well on a 12 well plate. Four sense strands and seven antisense strands were arranged in a matched matrix, as shown in Table 4 below. In Table 4 the references across the top right part of Table 4 (i.e., sense strand sequences) are the following: (1 ) s17_ori (SEQ ID NO: 8);(2) s17_allM (SEQ ID NO: 12); (3) s17_P3 (SEQ ID NO: 13); and (4) s17_P5 (SEQ ID NO: 14). In Table 4, the references across the left side of Table 4 {i.e., anti-sense strand sequences) are the following: (1) as_ori (SEQ ID NO: 2); (2) as_2 (SEQ ID NO: 15); (3) as_3 (SEQ ID NO: 16); (4) as_4 (SEQ ID NO: 17); (5) as_5 (SEQ ID NO: 18); (6) as_P3 (SEQ ID NO: 19) and (7) as_P5 (SEQ ID NO: 20). The references to the “shorthand” sequences on the left side of Table 4 refer to the following: (1) s17_ori (SEQ ID NO: 8);(2) s17_allM (SEQ ID NO: 12); (3) s17_P3 (SEQ ID NO: 13); (4) s17_P5 (SEQ ID NO: 14); (5) as_ori (SEQ ID NO: 2); (6) as_2 (SEQ ID NO: 15); (7) as_3 (SEQ ID NO: 16); (8) as_4 (SEQ ID NO: 17); (9) as_5 (SEQ ID NO: 18); (10) as_P3 (SEQ ID NO: 19) and (11) as_P5 (SEQ ID NO: 20. Each combination was passively transfected (e.g., 1 pM / 0.1 pM compounds, antibiotic-free EMEM medium 5% FBS, 48 hour incubation) into HeLA cells expressing a gene encoding a luciferase reporter. In these experiments, the level of knockdown was measured by Renilla luciferase expression, and normalized to constant Firefly luciferase expression. Data is expressed in FIG. 1A as the percentage of gene expression of non-transfected cells (NTC). In FIG. 1A, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 1 SEQ ID NO: 2); (2) s17_ori / as_2 (SEQ ID NO: 8 1 SEQ ID NO: 15); (3) s17_ori / as_3 (SEQ ID NO: 8 / SEQ ID NO: 16); (4) s17_ori / as_4 (SEQ ID NO: 8 / SEQ ID NO: 17); (5) s17_ori I as_5 (SEQ ID NO: 8 1 SEQ ID NO: 18); (6) s17_ori / as_P3 (SEQ ID NO: 8 / SEQ ID NO: 19; (7) s17_ori / as_P5 (SEQ ID NO: 81 SEQ ID NO: 20); (8) s17_allM I as_ori (SEQ ID NO: 12 / SEQ ID NO: 2); (9) s17_allM I as_2 (SEQ ID NO: 12 / SEQ ID NO: 15); (10) s17_allM I as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (11) s17_allM I as_4 (SEQ ID NO: 12 I SEQ ID NO: 17); (12) s17_allM I as_5 (SEQ ID NO: 12 / SEQ ID NO: 18); (13) s17_allM / as_P3 (SEQ ID NO: 12 / SEQ ID NO: 19); (14) s17_allM / as_P5 (SEQ ID NO: 12 / SEQ ID NO: 20); (15) s17_P3 1 as.ori (SEQ ID NO: 13 / SEQ ID NO: 2); (16) s17_P3 1 as_2 (SEQ ID NO: 13 / SEQ ID NO: 15); (17) s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (18) s17_P3 / as_4 (SEQ ID NO: 13 / SEQ ID NO: 17); (19) s17_P3 I as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (20) s17_P3 1 as_P3 (SEQ ID NO: 13 / SEQ ID NO: 19); (21) s17_P31 as_P5 (SEQ ID NO: 13 / SEQ ID NO: 20); (22) s17_P5 1 as_ori (SEQ ID NO: 14 / SEQ ID NO: 2); (23) s17_P5 1 as_2 (SEQ ID NO: 14 / SEQ ID NO: 15); (24) s17_P5 1 as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (25) s17_P5 / as_4 (SEQ ID NO: 14 / SEQ ID NO: 17); (26) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (27) s17_P5 / as_P3 (SEQ ID NO: 14 / SEQ ID NO: 19); and (28) s17_P5 / as_P5 (SEQ ID NO: 14 / SEQ ID NO: 20). In FIG. 1A, “VO” stands for “vector only.”

[0250] Statistical evaluation of the results is shown in FIG. 1B. In FIG. 1 B, the references in each column, starting from the left side of FIG. 1B, refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 / SEQ ID NO: 2); (2) s17_ori I as_2 (SEQ ID NO: 8 1 SEQ ID NO: 15); (3) s17_ori / as_3 (SEQ ID NO: 8 / SEQ ID NO: 16); (4) s17_ori / as_4 (SEQ ID NO: 8 / SEQ ID NO: 17); (5) s17_ori / as_5 (SEQ ID NO: 8 / SEQ ID NO: 18); (6) s17_ori I as_P3 (SEQ ID NO: 8 / SEQ ID NO: 19; (7) s17_ori / as_P5 (SEQ ID NO: 8 / SEQ ID NO: 20); (8) s17_allM I as_ori (SEQ ID NO: 12 / SEQ I D NO: 2); (9) s17_al I M I as_2 (SEQ I D NO: 12 / SEQ I D NO: 15); (10) s17_al I M / as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (11) s17_allM I as_4 (SEQ ID NO: 12 / SEQ ID NO: 17); (12) s17_allM I as_5 (SEQ ID NO: 12 / SEQ ID NO: 18); (13) s17_allM I as_P3 (SEQ ID NO: 12 / SEQ ID NO: 19); (14) s17_allM I as_P5 (SEQ ID NO: 12 / SEQ ID NO: 20); (15) s17_P3 / as_ori (SEQ ID NO: 13 / SEQ ID NO: 2); (16) s17_P3 1 as_2 (SEQ ID NO: 13 1 SEQ ID NO: 15); (17) s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (18) s17_P3 1 as_4 (SEQ ID NO: 13 / SEQ ID NO: 17); (19) s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (20) s17_P3 / as_P3 (SEQ ID NO: 13 / SEQ ID NO: 19); (21 ) s17_P3 / as_P5 (SEQ ID NO: 13 / SEQ ID NO: 20); (22) s17_P5 1 as_ori (SEQ ID NO: 14 / SEQ ID NO: 2); (23) s17_P5 / as_2 (SEQ ID NO: 14 / SEQ ID NO: 15); (24) s17_P5 / as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (25) s17_P5 I as_4 (SEQ ID NO: 14 / SEQ ID NO: 17); (26) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (27) s17_P5 1 as_P3 (SEQ ID NO: 14 / SEQ ID NO: 19); and (28) s17_P5 / as_P5 (SEQ ID NO: 14 / SEQ ID NO: 20). The experiments in this example demonstrate, inter alia, the dual dose screen of different modifications was successful in determining the effect of the patterns for the US36 target site. In these experiments, the four different sense strands did not significantly change the efficacy, however, the “AIIM” sense strand was observed to be less effective with all antisense strands. The seven antisense strands had a varying effect in different combinations with sense strands. s17_P3 1 as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); and s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18 were promising. In FIG. 1B, the black arrow points to sdRNA compounds that performed as well or better than the “original.”

[0251] A luciferase assay was developed to analyze dose curves and evaluate promising candidates (FIG. 2A and FIG. 2B) before experimental testing in cell lines and qPCR analysis. In FIG. 2A, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori I as_ori (SEQ ID NO: 8 1 SEQ ID NO: 2); (2) s17_ori / as_3 (SEQ ID NO: 8 I SEQ ID NO: 16); (3) s17_allM I as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (4) s17_P3 / as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (5) s17_P5 / as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (6) s17_ori / as_P5 (SEQ ID NO: 8 / SEQ ID NO: 20); (7) s17_P3 1 as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (8) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (9) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); and (10) US36_s_17 I US36_as (SEQ ID NO: 8 / SEQ ID NO: 2). In FIG. 2A, “NTC” stands for “Non-Template Control” (NTC_s I NTC_as (SEQ ID NO: 21 / SEQ ID NO: 22). In FIG. 2B, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) s17_ori / as_ori (SEQ ID NO: 8 1 SEQ ID NO: 2); (2) s17_ori I as_3 (SEQ ID NO: 8 / SEQ ID NO: 16); (3) s17_allM I as_3 (SEQ ID NO: 12 / SEQ ID NO: 16); (4) s17_P3 1 as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); (5) s17_P5 1 as_3 (SEQ ID NO: 14 / SEQ ID NO: 16); (6) s17_ori / as_P5 (SEQ ID NO: 8 / SEQ ID NO: 20); (7) s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); (8) s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18); (9) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); and (10) US36_s_17 1 US36_as (SEQ ID NO: 8 / SEQ ID NO: 2). In FIG. 2C, the references across the top of FIG. 2C ( / .e., sense strand sequences) are the following: (1) s17_ori (SEQ ID NO: 8);(2) s17_allM (SEQ ID NO: 12); (3) s17_P3 (SEQ ID NO: 13); and (4) s17_P5 (SEQ ID NO: 14). In FIG. 2C, the references across the left side of FIG. 2C ( / .e., anti-sense strand sequences) are the following: (1) as.ori (SEQ ID NO: 2); (2) as_2 (SEQ ID NO: 15); (3) as_3 (SEQ ID NO: 16); (4) as_4 (SEQ ID NO: 17); (5) as_5 (SEQ ID NO: 18); (6) as_P3 (SEQ ID NO: 19) and (7) as_P5 (SEQ ID NO: 20). As shown in FIG. 2A, there was not a significant difference observed in the alternative modification patterns tested in these dose curves; however, all of the combinations surprisingly had a 75-80% reduction in relative luciferase. FIG. 2C shows the best candidates from the screening. When comparing the I C50 values, none of the alternative modifications were better than the 71 pM value of US36_17 ori / ori ); ( / .e., s17_P3 1 as_3 (SEQ ID NO: 13 / SEQ ID NO: 16); s17_P3 / as_5 (SEQ ID NO: 13 / SEQ ID NO: 18); and s17_P5 / as_5 (SEQ ID NO: 14 / SEQ ID NO: 18). Thus, the experiments of this example demonstrate the alternative modification patterns of the sdRNA Compounds are effective in knocking down the USP10 target gene.

[0252] Example 2: Dose Curve Analysis of USP10 sdRNA Compounds with Alternative Modifications

[0253] In the experiments of this example, a reporter screening assay of USP10 sdRNA compounds was completed to determine if the dose curves were similar and representative of the dual does screen dose curves of the same compounds from Example 1 . In these experiments, about 10,000 HeLa cells were seeded per well on a 12 well plate. Ten USP10 sdRNA compounds varying in length from 12 to 20 nucleotides were passively transfected (e.g., 5 fold dose curve (1 pM - 0.000064 pM) compounds, antibiotic-free EMEM medium 5% FBS, 24 hour incubation) into HeLA cells expressing a gene encoding a luciferase reporter. In these experiments, knockdown was measured via Renilla luciferase expression and normalized to constant Firefly luciferase expression. Data is expressed in FIG. 3A as the percentage of gene expression of NTC. In FIG. 3A, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_12 I US36_as (SEQ ID NO: 4 I SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 1 SEQ ID NO: 2); (3) US36_s_14 / US36_as (SEQ ID NO: 6 1 SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 I SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 / SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 / SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36 old”) (SEQ ID NO: 1 I SEQ ID NO: 2). Statistical evaluation of the results is shown in FIG. 3B. In FIG. 3B, the references across the top of FIG. 3B refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_121 US36_as (SEQ ID NO: 4 / SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 I SEQ ID NO: 2); (3) US36_s_14 / US36_as (SEQ ID NO: 6 / SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 / SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 I SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 1 SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 1 SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36 old”) (SEQ ID NO: 1 I SEQ ID NO: 2). The experiments in this example demonstrate, inter alia, the dose curves were similar and representative of the dual does screen dose curves of the same compounds from Example 1 . The experiments of this example show how the differences between different sense lengths had minimal effect on overall maximal knockdown, but the differences did have an effect on the IC50 data. In these experiments, the US36_16 and US36_17 sdRNA compounds performed the best, and the US36_12 sdRNA compound performed the worst.

[0254] Example 3: Screening of USP 10 sdRNA Compounds (US36) with Different Sense Lengths

[0255] In the experiments of this example, a reporter screening assay of USP10 sdRNA compounds was completed to determine if different sense lengths affect the knockdown capacity of US36, as measured in a luciferase plasmid assay. In these experiments, about 10,000 HeLa cells were seeded per well on a 12 well plate. Ten USP10 sdRNA compounds varying in length from 12 to 20 nucleotides were passively transfected (e.g., 1 pM compounds, antibiotic-free EMEM medium 5% FBS, 24 hour incubation) into HeLA cells expressing a gene encoding a luciferase reporter. In these experiments, knockdown was measured via Renilla luciferase expression and normalized to constant Firefly luciferase expression. Data is expressed in FIG. 4 as the percentage of gene expression of NTC. In FIG. 4, the references across the X-axis refer to the following SEQ IDs as follows (from left to right; sense strand SEQ ID / antisense strand SEQ ID): (1) US36_s_12 I US36_as (SEQ ID NO: 4 / SEQ ID NO: 2); (2) US36_s_13 / US36_as (SEQ ID NO: 5 I SEQ ID NO: 2); (3) US36_s_141 US36_as (SEQ ID NO: 6 / SEQ ID NO: 2); (4) US36_s_15 / US36_as (SEQ ID NO: 1 1 SEQ ID NO: 2); (5) US36_s_16 / US36_as (SEQ ID NO: 7 / SEQ ID NO: 2); (6) US36_s_17 / US36_as (SEQ ID NO: 8 / SEQ ID NO: 2); (7) US36_s_18 / US36_as (SEQ ID NO: 9 1 SEQ ID NO: 2); (8) US36_s_19 / US36_as (SEQ ID NO: 10 / SEQ ID NO: 2); (9) US36_s_20 / US36_as (SEQ ID NO: 11 / SEQ ID NO: 2); and (10) US36_s_15 / US36_as (“US36” is same as“US36 old” in FIG. 3A and FIG. 3B) (SEQ ID NO: 1 / SEQ ID NO: 2). The experiments in FIG. 4 show that different sense lengths did not overall influence the knockdown of US36 as measured by the luciferase plasmid assay. In these experiments, US36_15 (with a 15mer sense) is the same as US36 from the last synthesis, and performed comparatively better. The only differences seen are with sense strands under 14mer, which had slightly decrease efficacy. Experiments evaluating full dose curves can show finer changes to the efficacy of knockdown.

[0256] Example 4: qPCR Analysis with Primary Cells In the experiments of this example, qPCR analysis was performed in primary human corneal fibroblast cells to analyze dose curves and evaluate promising candidates. The USP10 sdRNA compounds shown in Table 5, which vary in length from 12 to 20 nucleotides, were passively transfected (e.g., 2 pM compounds, DMEM / F12 + 10% FBS, 72 hour incubation) into primary human corneal fibroblast cells. In these experiments, gene expression was measured by qPCR (Taqman chemistry), and the data in FIG. 5A and FIG. 5B was adjusted to the standard curve, and normalized to the reference gene GAPDH. Data is expressed in FIG. 5A and FIG. 5B as the percentage of gene expression of non-transfected cells (NT). In these experiments, the dose curves of the US36 derivative siRNAs in primary human corneal fibroblast cells demonstrated RNAi inhibition of USP10. The best performing candidate in these experiments was US36_3 with about a 75% knockdown at the 2 uM dose and an IC50 of 0.935 uM, which is a greater knockdown than the original US36 compound (US36_15_Alt) as well as the parental compound US36 (sense 17 Alt).

[0257] Example 5: Efficacy and Safety of USP10-Targeted siRNA Treatments in Corneal Injury Models

[0258] The experiments of this example show how to conduct an in vivo corneal scrape procedure on C57BL / 6J mice to create a corneal injury for subsequent research. FIG. 6 shows an image of the in vivo corneal scraping procedure performed on C57BL / 6J mice (FIG. 6). The mice were first anesthetized using an intraperitoneal injection of ketamine, which was administered to ensure humane treatment and minimize discomfort during the intervention. To mark the area of corneal injury, a 2-mm circular trephine mark was utilized, which served as a guide for the subsequent scraping. A dulled blade, securely mounted on a blade breaker, was employed to execute a centripetal epithelial scrape. The arrows in FIG. 6 indicate the scrapping direction, which proceeds from the periphery toward the center of the cornea. Following this initial scraping, the same dulled blade was utilized to carefully collect and excise the corneal epithelium from the center of the cornea while ensuring the preservation of the underlying basement membrane, which is crucial for subsequent healing processes. The entirety of this procedure was conducted under a binocular microscope, ensuring precision in the execution of the corneal scrape. Aseptic conditions were strictly maintained throughout the procedure to minimize the risk of infection and to ensure the validity of the experimental outcomes.

[0259] The experiments in FIG. 7A, FIG. 7B, and FIG. 7C demonstrate the efficacy of US16 (SEQ ID NO: 571 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in preventing wound re-opening in a scrape- induced persistent corneal epithelial defect (PCED) model utilizing male mice at Day 28 post-injury. FIG. 7A shows a non-limiting schematic workflow of the experimental procedures undertaken in the scrape-induced PCED mouse model, providing a clear visual representation of the methodology employed. FIG. 7B shows representative images of fluorescein-stained corneas, demonstrating that treatments with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) effectively inhibited epithelial wound reopening four weeks following the PCED epithelial scrape. FIG. 7C provides quantification results obtained through analysis with the FIJI software, illustrating a statistically significant reduction in the open wound area within the US16 (SEQ ID NO: 57 I SEQ ID NO: 58) treatment group compared to the control groups (PBS and NTC (SEQ ID NO: 69 / SEQ ID NO: 70)). The data are derived from a cohort of 14 eyes per condition (N=14). The experiments in FIG. 7A, FIG. 7B, and FIG. 7C show the significant role of US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in promoting wound healing in the context of a scrape-induced PCED mouse model, highlighting their potential therapeutic applications in preventing corneal re-epithelialization failure.

[0260] The experiments in FIG. 8A, FIG. 8B, FIG. 8C, FIG. 8D, FIG. 8E, and FIG. 8F show the efficacy of US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 I SEQ ID NO: 60) on corneal scarring in male mice. FIG. 8A, FIG. 8B, FIG. 8C, FIG. 8D, FIG. 8E, and FIG. 8F shows graphs and images that examine the ability of US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 I SEQ ID NO: 60) to mitigate corneal scarring in male mice. FIG. 8A shows optical coherence tomography (OCT) imaging, which reveals the preservation of corneal structure following sdRNA treatment. FIG. 8B and FIG. 8C show quantification data on central corneal thickness, as measured using the FIJI software. The results confirm a restoration of corneal thickness to near-normal levels in drug-treated eyes, with sample sizes of N= 14 eyes for both the PBS and US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) treatment groups, and N=6 eyes for the NTC (SEQ ID NO: 69 / SEQ ID NO: 70) and unwounded controls. FIG. 8D shows measurements of central corneal thickness across the different treatment groups. FIG. 8E shows tonometry measurements that indicate no significant changes in intraocular pressure (IOP) among the treatment and control groups at Day 28. However, there is a discernible trend towards lower IOP in the groups treated with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 591 SEQ ID NO: 60), based on a sample size of N=8 eyes per condition. FIG. 8F shows the results of normalized fibrotic gene expression analysis conducted through RT-qPCR. The data indicate a decreasing trend in fibrotic gene expression in the US16 (SEQ ID NO: 57 / SEQ ID NO: 58)-treated group compared to the PBS control. The sample size for this analysis is N=8 eyes per condition, with RNA isolation conducted on pooled samples consisting of data from two eyes per sample. Collectively, the results shown in FIG. 8A, FIG. 8B, FIG. 8C, FIG. 8D, FIG. 8E, and FIG. 8F demonstrate the efficacy of US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 I SEQ ID NO: 60) in reducing corneal scarring, thereby contributing to the restoration of corneal integrity and thickness.

[0261] The experiments in FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, and FIG. 9E evaluate US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in reducing corneal scarring in female mice. FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, and FIG. 9E are graphs and images demonstrating the effectiveness of US16 (SEQ ID NO: 571 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 591 SEQ ID NO: 60) in reducing corneal scarring in female mice. In conjunction with data collected from male mice, the therapeutic effects of USP10-targeted siRNA (US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) were further assessed in female mice utilizing the persistent corneal epithelial defect (PCED) scrape model. FIG. 9A shows that, similar to what was observed in male mice, the corneal epithelium in female mice at Day 28 exhibited recurrent wound re-opening and corneal abrasions in the control groups (PBS and NTC (SEQ ID NO: 69 1 SEQ ID NO: 70)). In contrast, the drug-treated groups (US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) showed no signs of re-opening. Specifically, 9 out of 10 corneas in the drug-treated groups demonstrated no reopening, compared to just 1 out of 10 in the control groups. FIG. 9C shows that optical coherence tomography (OCT) imaging at Day 28 revealed similar corneal morphology in drug-treated female mice as observed in male mice, characterized by intact epithelium and endothelium and improved morphology following sdRNA treatment. These experiments show that US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) significantly reduce corneal scarring in female mice, mirroring the therapeutic effects observed in male mice, and providing further evidence for the effectiveness of USP10-targeted siRNA in promoting corneal healing.

[0262] The experiments in FIG. 10A and FIG. 10B assess corneal cell morphology and endothelial cell density following US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatment. FIG. 10A and FIG. 10B show analysis of corneal cell morphology and endothelial cell density, demonstrating that treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) does not adversely affect these parameters in male mice, suggesting that both treatments are non-toxic. FIG. 10A shows in vivo confocal images that reveal no differences in the size, shape, or morphology of endothelial cells across both treated and control groups. Additionally, the stromal keratocytes in all groups consistently display their normal organized patterns. FIG. 10B shows a graph detailing the analysis of endothelial cell density conducted using HEYEX software. The results indicate that there are no significant changes in endothelial cell density in the US16 (SEQ ID NO: 57 / SEQ ID NO: 58)-treated groups when compared to the PBS control and unwounded groups, with a sample size of N=4 eyes per condition. The experiments in FIG. 10A and FIG. 10B support the findings that US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatment does not compromise corneal morphology or endothelial cell density in male mice, thereby supporting their non-toxic profile.

[0263] The experiments in FIG. 11 A, FIG. 11 B, and FIG. 11 C show histological evaluation of corneal morphology following treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in male mice. FIG. 11 A, FIG. 11 B, and FIG. 11 C show hematoxylin and eosin (H&E) images collected after wounding and subsequent treatment with US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 591 SEQ ID NO: 60) in male mice. The H&E images indicate that overall corneal morphology showed significant improvement in the US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatment groups, with no observed signs of corneal erosions. Both the corneal epithelium and stroma were restored to their normal structural configuration following treatment. Notably, the results from the experiments in female mice (FIG. 12A, FIG. 12B, and FIG. 12C) corroborate the findings in these male mice experiments of FIG. 11 A, FIG. 11 B, and FIG. 11C, suggesting that there are no significant sex-based differences in the efficacy of USP10-targeted siRNA treatments in preventing wound reopening and promoting corneal healing after epithelial scraping.

[0264] The experiments in FIG. 12A, FIG. 12B, and FIG. 12C show histological evaluation of corneal morphology following treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in female mice. FIG. 12A, FIG. 12B, and FIG. 12C show the overall corneal morphology exhibited notable improvement in the US16 (SEQ ID NO: 571 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) treatment groups, with no indications of corneal erosions. Both the corneal epithelium and stroma were restored to their normal structural configuration following treatment. The results observed in these experiments with female mice (FIG. 12A, FIG. 12B, and FIG. 12C) are consistent with those obtained from male mice (FIG. 11 A, FIG. 11 B, and FIG. 11 C), suggesting thatthere are no significant sex-based differences in the efficacy of USP10-targeted siRNA treatments for preventing wound reopening and enhancing corneal healing after epithelial scraping.

[0265] The experiments in FIG. 13 show immunohistochemical analysis of hemidesmosomes following treatment with US16 (SEQ ID NO: 57 1 SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60) in male mice. FIG. 13 shows immunohistochemistry images depicting hemidesmosome structures following wounding and subsequent treatment with US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60) in male mice. Twenty-eight days post-wounding and drug treatment, the mice were sacrificed, and immunostaining was performed for Collagen XVII, a critical marker for hemidesmosomes that facilitate the adhesion of the epithelium to the basement membrane in the cornea. All experimental conditions exhibited Collagen XVII staining indicative of hemidesmosome reformation. Notably, the epithelium in the sdRNA- treated corneas displayed stratification with qualitatively improved intercalation into the epithelial layers compared to the PBS-treated corneas. The sample size for this analysis is N=4 eyes.

[0266] The experiments in FIG. 14A, FIG. 14B, FIG. 14C, FIG. 14D, and FIG. 14E show a safety assessment of USP10 knockdown via USP10-targeting siRNA (US09 (SEQ ID NO: 61 1 SEQ ID NO: 62)) following epithelial scratch injury in New Zealand Rabbits. Female and male New Zealand rabbits were subjected to a light scratch wound to the central cornea, ensuring that the basement membrane remained intact. Over a period of 3.5 months, corneas were analyzed from three groups: PBS, US09 (SEQ ID NO: 61 I SEQ ID NO: 62), and US09.1 (SEQ ID NO: 63 / SEQ ID NO: 64). Day 0 refers to the time immediately after wounding. Clinical ophthalmologic evaluations conducted at regular intervals throughout the 3.5 months revealed no significant differences among the eyes in the three groups across any of the assessed parameters. These periodic evaluations indicate that siRNA treatment was safe and tolerable for rabbit eyes in vivo. These experiments demonstrated that both US09 (SEQ ID NO: 61 / SEQ ID NO: 62) and US09.1 (SEQ ID NO: 63 / SEQ ID NO: 64) are non-toxic to the rabbit cornea. The following parameters were not significantly different among the groups: Central corneal thickness (FIG. 14A), Intraocular pressure (IOP) (FIG. 14B), Schirmer tear strips (FIG. 14C), and weight (FIG. 14D). Imaging assessments of eye health showed no signs of ocular inflammation, corneal haze, neovascularization, chemosis, or any other ocular abnormalities affecting the cornea, conjunctiva, or sclera following sdRNA treatment (FIG. 14E).

[0267] The experiments in FIG. 15A, FIG. 15B, and FIG. 15C show another safety profile of USP10 knockdown using USP10-targeting siRNA (US09 (SEQ ID NO: 61 / SEQ ID NO: 62)) following an epithelial scratch injury in female and male New Zealand rabbits. In FIG. 15A, optical coherence tomography (OCT) analyses revealed no differences in the optical properties of the corneas treated with US09 (SEQ ID NO: 61 1 SEQ ID NO: 62) or US09.1 (SEQ ID NO: 63 / SEQ ID NO: 64) when compared to unwounded rabbit corneas. In FIG. 15B, representative confocal images acquired using HRT3 technology indicated that there were no discernible variations in the morphology of each layer of the rabbit cornea. The stromal layers exhibited an organized keratocyte pattern with observable basal nerve plexuses. The examples presented are from female rabbits, and no sex-dependent differences were noted in the analyses. In FIG. 15C, examination of the endothelial layer across all groups overthe duration of 3.5 months demonstrated a normal hexagonal pattern, indicating that there was no evidence of toxicity associated with either US09 (SEQ ID NO: 61 / SEQ ID NO: 62) or US09.1 (SEQ ID NO: 63 1 SEQ ID NO: 64). These experiments support the conclusion that USP10- targeting siRNA (US09 (SEQ ID NO: 61 1 SEQ ID NO: 62)) is safe for use following epithelial scratch injury. The lack of differences in corneal morphology, structured organization within the stromal layers, and the maintenance of a normal endothelial pattern indicate that both US09 (SEQ ID NO: 61 / SEQ ID NO: 62) and US09.1 (SEQ ID NO: 631 SEQ ID NO: 64) do not induce any toxic effects. These results highlight the potential of USP10-targeted siRNA as a non-toxic therapeutic option in ocular applications.

[0268] The experiments in FIG. 16 show the evaluation of USP10 knockdown with US36.1 (SEQ ID NO: 67 1 SEQ ID NO: 68) in ex vivo human corneas. Cadaver corneas were obtained and an 8 mm trephine was employed to demarcate the wound margins, positioned just inside the limbus. Using a centripetal motion, the epithelial layer was scraped away, leaving the basement membrane intact. The corneas were subsequently mounted on an agar / collagen base and placed into a 60 mm dish containing supplemented serum-free media that reached up to the limbus, thereby maintaining an air / liq uid interface with the corneal surface. Central corneal regions received either one dose of PBS or a treatment of 12.4 pg of US36.1 (SEQ ID NO: 67 1 SEQ ID NO: 68) / DUB-002 (SEQ ID NO: 67 / SEQ ID NO: 68) in 35 μl. The corneas were subsequently moistened with the supplemented serum-free media 24 hours after culture initiation and every two days thereafter. At the two-week mark, the corneas were bisected through the wound, and one half was fixed in 10% formalin for paraffin embedding and hematoxylin and eosin (H&E) staining. The bar scale in FIG. 16 represents 100 pm. The donor age for this sample was 66 years. The results indicated that US36.1 (SEQ ID NO: 67 / SEQ ID NO: 68) significantly promoted regenerative healing in the epithelium at the two-week mark compared to the controls treated with PBS, and the results also demonstrate that USP10 knockdown using US36.1 (SEQ ID NO: 67 I SEQ ID NO: 68) effectively enhances epithelial regeneration in ex vivo human corneas after two weeks. This suggests that US36.1 (SEQ ID NO: 67 / SEQ ID NO: 68) has potential therapeutic applications in promoting corneal healing, warranting further investigations into its efficacy in clinical settings.

[0269] The nomenclature for sdRNAs is as follows: US16 (SEQ ID NO: 57 1 SEQ ID NO: 58), US09 (SEQ ID NO: 61 / SEQ ID NO: 62), US36 (SEQ ID NO: 65 / SEQ ID NO: 66), and NTC (SEQ ID NO: 69 1 SEQ ID NO: 70) represent the parent sequences 20 / 15 along with their corresponding internal nomenclature. (See Table 6) DUB-001 refers to the human test article (SEQ ID NO: 65 1 SEQ ID NO: 66). The sequences US16.1 (SEQ ID NO: 571 SEQ ID NO: 58), US09.1 (SEQ ID NO: 63 1 SEQ ID NO: 64), and US36.1 (SEQ ID NO: 67 / SEQ ID NO: 68) denote novel 20 / 17 sequences that incorporate a new pattern of modifications. DUB-002 (SEQ ID NO: 671 SEQ ID NO: 68) is identified as the novel human test article.

[0270] Collectively, these experiments show that the USP10-targeted siRNA treatments (e.g., US16 (SEQ ID NO: 57 / SEQ ID NO: 58) and US16.1 (SEQ ID NO: 59 / SEQ ID NO: 60), demonstrate significant efficacy in promoting corneal healing and preventing wound re-opening in both male and female mouse models of persistent corneal epithelial defects. Additionally, the treatments are non-toxic, as evidenced by the preservation of corneal morphology and endothelial density, suggesting their safety for potential therapeutic use. Furthermore, the results observed with US36.1 (SEQ ID NO: 67 / SEQ ID NO: 68) in ex vivo human corneas indicate a promising avenue for enhancing epithelial regeneration, supporting further investigation into its clinical applications.

[0271] The present disclosure demonstrates synthesis and biological activity of the chemically synthesized sdRNA compounds containing various modifications for RNAi to inhibit expression of USP10. Without wishing to be bound by theory, the use of these modifications enhances the stability of the sdRNAs and the utility as a therapeutic agent.

[0272] As used herein, the word “include,” and its variants, is intended to be non-limiting, such that recitation of items in a list is not to the exclusion of other like items that may also be useful in the materials, compositions, devices, and methods of this technology. Similarly, the terms “can” and “may” and their variants are intended to be non-limiting, such that recitation that an embodiment can or may comprise certain elements or features does not exclude other embodiments of the present technology that do not contain those elements or features. Although the open-ended term “comprising,” as a synonym of terms such as including, containing, or having, is used herein to describe and claim the disclosure, the present technology, or embodiments thereof, may alternatively be described using more limiting terms such as “consisting of’ or “consisting essentially of” the recited ingredients.

[0273] Unless defined otherwise, all technical and scientific terms herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials, similar or equivalent to those described herein, can be used in the practice or testing of the present disclosure, the preferred methods and materials are described herein.

[0274] All publications, patents, and patent publications cited are incorporated by reference herein in their entirety for all purposes. All of the features disclosed herein may be combined in any combination. Each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0275] From the above description, one skilled in the art can easily ascertain the essential characteristics of the present disclosure, and without departing from the spirit and scope thereof, can make various changes and modifications of the disclosure to adapt it to various usages and conditions. Thus, other embodiments are also within the claims.

[0276] EMBODIMENTS

[0277] Various additional embodiments of the disclosure are provided by the following enumerated embodiments, which can be combined in any number and in any combination.

[0278] Embodiment 1. A composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, and the sense strand is about 16 to about 18 nucleotides in length and has sequence complementarity to the antisense strand.

[0279] Embodiment 2. The composition of embodiment 1 , wherein the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition.

[0280] Embodiment 3. The composition of embodiment 1 , wherein the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition.

[0281] Embodiment 4. The composition of embodiment 1 , wherein the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition.

[0282] Embodiment 5. The composition of embodiment 1 , wherein the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. Embodiment 6. The composition of any one of embodiments 1-5, wherein the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

[0283] Embodiment 7. The composition of any one of embodiments 1-5, wherein the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

[0284] Embodiment 8. The composition of any one of embodiments 1-7, wherein the sense strand is about 16 nucleotides in length.

[0285] Embodiment 9. The composition of any one of embodiments 1-7, wherein the sense strand is about 17 nucleotides in length.

[0286] Embodiment 10. The composition of any one of embodiments 1-7, wherein the sense strand is about 18 nucleotides in length.

[0287] Embodiment 11. The composition of any one of embodiments 1-7, wherein the nucleic acid complex has: a sense strand consisting of 16 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

[0288] Embodiment 12. The composition of any one of embodiments 1-7, wherein the nucleic acid complex has: a sense strand consisting of 17 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

[0289] Embodiment 13. The composition of any one of embodiments 1-7, wherein the nucleic acid complex has: a sense strand consisting of 18 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

[0290] Embodiment 14. The composition of any one of embodiments 1-7, wherein the sense strand comprises the base sequence of SEQ ID NOs :1 -2, or a variant thereof.

[0291] Embodiment 15. The composition of any one of embodiments 1-7 or 14, wherein the antisense strand comprises the base sequence of SEQ ID NO: 3 or a variant thereof. Embodiment 16. The composition of any one of embodiments 14-15, wherein the variant comprises about 1 , or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0292] Embodiment 17. The composition of any one of embodiments 1 -7, wherein the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 6, or a variant thereof.

[0293] Embodiment 18. The composition of any one of embodiments 1-7 or 17, wherein the antisense strand consists of the base sequence of SEQ ID NO: 7, or a variant thereof.

[0294] Embodiment 19. The composition of any one of embodiments 1-18, wherein the nucleic acid complex comprises one or more chemical modifications.

[0295] Embodiment 20. The composition of embodiment 19, wherein the chemical modification is a 2'-O-methylated nucleoside, a phosphorothioate bond, or a hydrophobic moiety.

[0296] Embodiment 21. The composition of any one of embodiments 19-20, wherein the chemical modification is selected from a locked nucleic acid (LNA), phosphorothioate, 2'-O-Methyl, 2'-O-Methoxyethyl, 2'-O-alkyl-RNA unit, 2'-OMe-RNA unit, 2 -amino-DNA unit, 2'-fl uoro-DNA unit, peptide nucleic acid (PNA) unit, hexitol nucleic acids (HNA) unit, INA unit, and a 2'-O-(2-Methoxyethyl)-RNA (2' MOE RNA) unit.

[0297] Embodiment 22. The composition of any one of embodiments 19-20, wherein the chemical modification is a 2' fluoro group.

[0298] Embodiment 23. The composition of any one of embodiments 19-20, wherein the chemical modification is 2'-O-methyl (2'OMe) group.

[0299] Embodiment 24. The composition of any one of embodiments 19-20, wherein the chemical modification is a phosphorothioate linker.

[0300] Embodiment 25. The composition of any one of embodiments 19-21, wherein the chemical modification is a 5'-vinylphosphonate 2'O-mll group.

[0301] Embodiment 26. The composition of any one of embodiments 19-20, wherein the chemical modification is a cholesterol conjugate, or the chemical modification comprises docosanoic acid (DCA). Embodiment 27. The composition of any one of embodiments 1-26, wherein the composition comprises a modified sequence selected from:

[0302] (a) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 9) 5'

[0303] [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 10) 5'

[0304] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][ Ps][mG][Ps][fC][Ps][mC][Ps][fG] 3';

[0305] (b) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 11) 5'

[0306] [fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 12) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][ Ps][fC][Ps][mC][Ps][fG] 3';

[0307] (c) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 13) 5'

[0308] [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'Ch olTEG] 3'; and an antisense strand of: (SEQ ID NO: 14) 5'

[0309] [5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][ Ps][mG][Ps][fC][Ps][mC][Ps][fG] 3'; and

[0310] (d) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 15) 5'

[0311] [fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'Ch olTEG] 3'; and an antisense strand of: (SEQ ID NO: 16) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][ Ps][fC][Ps][mC][Ps][fG] 3'; wherein f represents 2' Fluoro, m represents 2' O-methyl, Ps represents phosphorothioate linker, 5Phos represents 5' phosphate, 3'CholTEG represents Cholesterol conjugate, and vP-mll represents 5'- Vinylphosphonate 2'0-mll.

[0312] Embodiment 28. A composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, the sense strand is about 15 nucleotides in length and having sequence complementarity to the antisense strand; and the antisense strand and the sense strand comprise one or more chemical modifications, selected from P, #, f, 2’ Fluor, m, 2’0-methyl, Choi 3’, cholesterol and docosanoic acid (DCA), wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0313] Embodiment 29. The composition of embodiment 28, wherein the composition is suitable for causing a reduction of expression and / or activity of a USP10 mRNA, as compared to expression and / or activity in the absence of the composition.

[0314] Embodiment 30. The composition of embodiment 28, wherein the composition is suitable for causing a reduction of expression and / or activity of a USP10 protein, as compared to expression and / or activity in the absence of the composition.

[0315] Embodiment 31. The composition of embodiment 28, wherein the composition is suitable for preventing or reducing an upregulation of a USP10 mRNA, as compared to upregulation in the absence of the composition.

[0316] Embodiment 32. The composition of embodiment 28, wherein the composition is suitable for preventing or reducing an upregulation of a USP10 protein, as compared to upregulation in the absence of the composition. Embodiment 33. The composition of any one of embodiments 28-32, wherein the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

[0317] Embodiment 34. The composition of any one of embodiments 28-32, wherein the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

[0318] Embodiment 35. The composition of any one of embodiments 28-34, wherein the nucleic acid complex has: a sense strand consisting of 15 nucleotides in length and an antisense strand consisting of 20 nucleotides in length.

[0319] Embodiment 36. The composition of any one of embodiments 28-35, wherein the sense strand comprises the base sequence of SEQ ID NO: 5 or a variant thereof.

[0320] Embodiment 37. The composition of any one of embodiments 28-36, wherein the antisense strand comprises the base sequence of SEQ ID NO: 3, or a variant thereof.

[0321] Embodiment 38. The composition of any one of embodiments 36-37, wherein the variant comprises about 1 , or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

[0322] Embodiment 39. The composition of any one of embodiments 28-36, wherein the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof.

[0323] Embodiment 40. The composition of any one of embodiments 28-37, wherein the antisense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof.

[0324] Embodiment 41. The composition of any one of embodiments 28-40, wherein the composition comprises a modified sequence selected from a nucleic acid complex comprising: a sense strand selected from

[0325] 5' mC#mA#mG.mA.mC.mC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 17);

[0326] 5' mC#mA#mG.mG.mA.mA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 18);

[0327] 5' mT#mA#mT.mT.mC.mT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 19);

[0328] 5' mC#mA#mG.mA.fC.fC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 20);

[0329] 5' mC#mA#mG.mG.fA.fA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 21); 5' mT#mA#mT.mT.fC.fT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 22);

[0330] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 23);

[0331] 5' mC#rnA#mG.mG.fA.fA.mA.mG.fA.rnC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 24);

[0332] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 25);

[0333] 5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.fA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 26);

[0334] 5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.fT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 27);

[0335] 5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.fr.mA.mA#mT#mA-Chol 31(SEQ ID NO: 28);

[0336] 5' mC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#mA-Chol 3' (SEQ ID NO: 29);

[0337] 5' mC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#mA-Chol 3' (SEQ ID NO: 30);

[0338] 5' mT#mA#fT.rnT.fC.mT.fT.mG.fG.mT.fT.mA.fA#mT#niA-Chol 3' (SEQ ID NO: 31);

[0339] 5' fC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#fA-Chol 3' (SEQ ID NO: 32);

[0340] 5' fC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#fA-Chol 3' (SEQ ID NO: 33);

[0341] 5' fT#mA#fT.mT.fC.mT.fr.mG.fG.mT.fr.mA.fA#mT#fA-Chol 3' (SEQ ID NO: 34); and an antisense strand of:

[0342] 5' PmT#fC#mT.mT.mT.mC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 35);

[0343] 5' mT#fA#mT.mC.mA.mG.mT.mC.mT.mT.mT.mC.nnC.fT#mG#nnG#mT#mG#mA#mG 3' (SEQ ID NO: 36);

[0344] 5'PmT#fA#mT.mT.mA.mA.mC.mC.mA.mA.mG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 37);

[0345] 5' PmT#fC#mT.mT.mT.fC.mT.mT.mT.mG.mG.mT.mC.fr#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 38);

[0346] 5' PmT#fA#mT.mC.mAJG.mT.mC.mT.mT.mT.mC.mCK#mG#mG#mT#mG#mA#mG 3' (SEQ ID

[0347] NO: 39);

[0348] 5'PmT#fA#mT.mT.mA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 40);

[0349] 5'PmT#fC#mT.mT.mT.mC.mT.mT.mT.fG.fG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 41);

[0350] 5' PmT#fA#mT.mC.mA.mG.mT.mC.mT.fT.1T.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 42); 5' PmT#fA#mT.mT.mA.mA.mC.mC.mA.fA.fG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0351] 43);

[0352] 5' PmT#fC#mT.fT.fT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:

[0353] 44);

[0354] 5' PmT#fA#mT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:45);

[0355] 5' PmT#fA#mT.fr.fA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0356] 46);

[0357] 5' PmT#fC#n’.fT.fr.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:

[0358] 47);

[0359] 5' PmT#fA#fT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:

[0360] 48);

[0361] 5' PmT#fA#fT.fT.fA.fA.mC.mC.mA.mA.mG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:

[0362] 49);

[0363] 5' PmT#fC#mT.fr.fT.fC.fr.fT.mT.fG.mG.mT.mC.fr#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 50);

[0364] 5' PmT#fA#mT.fC.fA.fG.fT.fC.mT.fr.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 51);

[0365] 5' PmT#fA#mT.1T.fA.fA.fC.fC.mA.fA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 52);

[0366] 5' PmT#fC#mT.fr.mT.fC.mT.fr.mT.fG.mG.fT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 53);

[0367] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fr.mT.fC.mC.fr#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:54);

[0368] 5' PmT#fA#mT.fT.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 55);

[0369] 5' PmT#fC#mT.fT.mT.fC.mT.tT.mT.fG.mG.fT.mC.fT#mG#fC#mA#IT#mT#fC 3' (SEQ ID NO: 56);

[0370] 5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fT.mT.fC.mC.fT#mG#fG#mT#fG#mA#fG 3' (SEQ ID NO: 57);

[0371] 5' PmT#fA#mT.1T.mA.fA.mC.fC.mA.fA.mG.fA.mA.1T#mA#fC#mT#fG#mA#fA 3' (SEQ ID NO: 58); wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

[0372] Embodiment 42. A pharmaceutical composition comprising the composition of any one of embodiments 1- 41 , and a pharmaceutically acceptable carrier. Embodiment 43. The pharmaceutical composition of embodiment 42, wherein the pharmaceutical composition is suitable for self-delivery.

[0373] Embodiment 44. The pharmaceutical composition of any one of embodiments 42-43, further comprising a vehicle.

[0374] Embodiment 45. The pharmaceutical composition of any of one embodiments 42-44, wherein the vehicle is one or more of calcium phosphate, a cationic lipid, a cationic polymer, polyethyleneimine and a protein-based transfection reagent.

[0375] Embodiment 46. The pharmaceutical composition of any one of embodiments 42-45, wherein the composition is formulated for topical, pulmonary, or parenteral delivery.

[0376] Embodiment 47. A cell comprising the composition or pharmaceutical composition of any one of embodiments 1-46.

[0377] Embodiment 48. A method of promoting or enhancing degradation of an mRNA encoding USP10, comprising: administering to a subject in need thereof an effective amount of the composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, or contacting a cell with the composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46 and administering to a subject in need thereof an effective amount of the composition, wherein the promoting or enhancing is as compared to a state before or without composition.

[0378] Embodiment 49. The method of embodiment 48, wherein the method is performed in vitro.

[0379] Embodiment 50. The method of embodiment 48, wherein the method is performed ex vivo.

[0380] Embodiment 51. The method of embodiment 48, wherein the method is performed in vivo.

[0381] Embodiment 52. A method of treating a disease or disorder, comprising: administering to a subject in need thereof an effective amount of the composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, or contacting a cell with the composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46 and administering to a subject in need thereof an effective amount of the composition.

[0382] Embodiment 53. The method of embodiment 52, wherein the disease or disorder is characterized by scarring.

[0383] Embodiment 54. The method of embodiment 53, wherein the disease or disorder is selected from ocular scarring (optionally selected from scarring of the cornea and / or retina), integumentary scarring, internal organ scarring, and internal organ (optionally selected from lung, liver, and kidney) fibrosis.

[0384] Embodiment 55. A method of reducing or eliminating ocular scarring in an eye of a subject after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0385] Embodiment 56. A method for accelerating wound closure in an eye of a subject after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the accelerating is compared to upregulation without treatment or pre-treatment.

[0386] Embodiment 57. A method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of a subject after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the suppressing is compared to upregulation without treatment or pre-treatment.

[0387] Embodiment 58. A method of eliminating or reducing fibrosis of a subject after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0388] Embodiment 59. A method of eliminating or reducing scarring in a skin of a subject after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0389] Embodiment 60. A method of eliminating or reducing scarring in an eye of a subject as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of embodiments 1 -41 or pharmaceutical composition of any one of embodiments 42- 46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment.

[0390] Embodiment 61. The method of any one of embodiments 48-60, wherein the composition is administered at a frequency of no more than about once monthly, no more than about once every two months.

[0391] Embodiment 62. The method of any one of embodiments 48-61 , further comprising administering one or more of an antibiotic and a steroid agent.

[0392] Embodiment 63. The method of any one of embodiments 48-61 , wherein the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0393] Embodiment 64. The method of any one of embodiments 48-63, wherein the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment.

[0394] Embodiment 65. The method of any one of embodiments 48-64, wherein the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0395] Embodiment 66. The composition of any one of embodiments 1-41 , or the pharmaceutical composition of any one of embodiments 42-46, or the cell of embodiment 47, or the method of any one of embodiments 48- 65, wherein the USP10 mRNA or protein is from a mammal, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0396] Embodiment 67. A method of reducing or eliminating ocular scarring in an eye of an animal after an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human. Embodiment 68. A method for accelerating wound closure in an eye of an animal after an ocular wound comprising administering to the wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the accelerating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0397] Embodiment 69. A method for suppressing a production of fibrotic markers in a tissue after a wound or immune response in an eye of an animal after an ocular wound, comprising administering to the wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the suppressing is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0398] Embodiment 70. A method of eliminating or reducing fibrosis of an animal after a tissue wound comprising administering to the tissue wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0399] Embodiment 71. A method of eliminating or reducing scarring in a skin of an animal after a skin wound, comprising administering to the skin wound a therapeutically effective amount of a composition of any one of embodiments 1-41 or pharmaceutical composition of any one of embodiments 42-46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0400] Embodiment 72. A method of eliminating or reducing scarring in an eye of an animal as a result of healing of an ocular wound comprising administering to the ocular wound a therapeutically effective amount of a composition of any one of embodiments 1 -41 or pharmaceutical composition of any one of embodiments 42- 46, wherein the reducing or eliminating is compared to upregulation without treatment or pre-treatment, optionally wherein the mammal is selected from a monkey, cat, dog, horse, donkey, sheep, pig, goat, cow, mouse, rat, rabbit, guinea pig, and human.

[0401] Embodiment 73. The method of any one of embodiments 67-72, wherein the composition is administered at a frequency of no more than about once monthly, no more than about once every two months. Embodiment 74. The method of any one of embodiments 67-73, further comprising administering one or more of an antibiotic and a steroid agent.

[0402] Embodiment 75. The method of any one of embodiments 67-73, wherein the administering obviates the need for administration of one or more of an antibiotic and a steroid agent.

[0403] Embodiment 76. The method of any one of embodiments 67-75, wherein the method substantially eliminates an upregulation of USP10 mRNA or protein, as compared to upregulation without treatment or pre-treatment.

[0404] Embodiment 77. The method of any one of embodiments 67-76, wherein the method substantially reduces or eliminates vision loss, as compared to upregulation without treatment or pre-treatment.

[0405] Table 4: Matched Matrix of Sense and Antisense Strands

[0406] Table 5: Sense and Antisense References for FIG. 5A and FIG. 5B

[0407] Table 6. Nomenclature of sdRNAs. US16 (SEQ ID NO: 57 / SEQ ID NO: 58), US09 (SEQ ID NO: 61 1 SEQ ID NO: 62), US36 (SEQ ID NO: 65 / SEQ ID NO: 66), NTC (SEQ ID NO: 69 / SEQ ID NO: 70) are the parent 20 / 15 sequences with internal nomenclature. DUB-001 is a human test article (SEQ ID NO: 65 / SEQ ID NO: 66). US16.1 (SEQ ID NO: 59 1 SEQ ID NO: 60), US09.1 (SEQ ID NO: 63 1 SEQ ID NO: 64), US36.1 (SEQ

[0408] ID NO: 67 / SEQ ID NO: 68) are the novel 20 / 17 sequences with novel pattern of modifications. DUB-002 is a novel human test article (SEQ ID NO: 67 / SEQ ID NO: 68).

Claims

CLAIMSWhat is claimed is:

1. A composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, and the sense strand is about 16 to about 18 nucleotides in length and has sequence complementarity to the antisense strand.

2. The composition of claim 1 , wherein the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

3. The composition of claim 1 , wherein the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

4. The composition of any one of claims 1 -3, wherein the nucleic acid complex has: a sense strand consisting of 16 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

5. The composition of any one of claims 1 -3, wherein the nucleic acid complex has: a sense strand consisting of 17 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

6. The composition of any one of claims 1 -3, wherein the nucleic acid complex has: a sense strand consisting of 18 nucleotides in length, and an antisense strand consisting of 20 nucleotides in length.

7. The composition of any one of claims 1-3, wherein the sense strand comprises the base sequence of SEQ ID NOs :1 -2, or a variant thereof.

8. The composition of any one of claims 1-3 or 7, wherein the antisense strand comprises the base sequence of SEQ ID NO: 3 or a variant thereof.

9. The composition of any one of claims 7-8, wherein the variant comprises about 1 , or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

10. The composition of any one of claims 1-3, wherein the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 6, or a variant thereof.

11. The composition of any one of claims 1-3 or 10, wherein the antisense strand consists of the base sequence of SEQ ID NO: 7, or a variant thereof.

12. The composition of any one of claims 1-11 , wherein the nucleic acid complex comprises one or more chemical modifications.

13. The composition of claim 12, wherein the chemical modification is a 5'-vinylphosphonate 2'0-mU group.

14. The composition of any one of claims 1-13, wherein the composition comprises a modified sequence selected from:(a) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 9) 5'[fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 10) 5'[5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][ Ps][mG][Ps][fC][Ps][mC][Ps][fG] 3';(b) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 11) 5'[fA][Ps][mA][Ps][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3-CholTEG] 3'; and an antisense strand of: (SEQ ID NO: 12) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][ Ps][fC][Ps][mC][Ps][fG] 3';(c) a nucleic acid complex comprising:a sense strand of (SEQ ID NO: 13) 5'[fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3’Ch olTEG] 3'; and an antisense strand of: (SEQ ID NO: 14) 5'[5Phos][mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][ Ps][mG][Ps][fC][Ps][mC][Ps][fG] 3'; and(d) a nucleic acid complex comprising: a sense strand of (SEQ ID NO: 15) 5'[fC][Ps][mU][Ps][fA][mA][fU][mG][fA][mA][fU][mG][fA][mG][fU][mU][fC][Ps][mA][Ps][fA][3'Ch olTEG] 3'; and an antisense strand of: (SEQ ID NO: 16) 5' [vP- mU][Ps][fU][Ps][mG][fA][mA][fC][mU][fC][mA][fU][mU][fC][mA][fU][Ps][mU][Ps][fA][Ps][mG][ Ps][fC][Ps][mC][Ps][fG] 3'; wherein f represents 2' Fluoro, m represents 2' O-methyl, Ps represents phosphorothioate linker, 5Phos represents 5' phosphate, 3'CholTEG represents Cholesterol conjugate, and vP-mll represents 5'- Vinylphosphonate 2'0-mll.

15. A composition comprising a nucleic acid complex for inducing RNA interference (RNAi) to inhibit expression of a ubiquitin specific peptidase 10 (USP10) gene, the nucleic acid complex comprising an antisense strand and a sense strand, wherein: the antisense strand is about 20 nucleotides in length, the sense strand is about 15 nucleotides in length and having sequence complementarity to the antisense strand; and the antisense strand and the sense strand comprise one or more chemical modifications, selected from P, #, f, 2’ Fluor, m, 2’0-methyl, Choi 3’, cholesterol and docosanoic acid (DCA), wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

16. The composition of claim 15, wherein the nucleic acid complex has perfect sequence complementarity to an mRNA molecule encoding USP10.

17. The composition of claim 15, wherein the nucleic acid complex has partial sequence complementarity to an mRNA molecule encoding USP10.

18. The composition of any one of claims 15-17, wherein the nucleic acid complex has: a sense strand consisting of 15 nucleotides in length and an antisense strand consisting of 20 nucleotides in length.

19. The composition of any one of claims 15-18, wherein the sense strand comprises the base sequence of SEQ ID NO: 5 or a variant thereof.

20. The composition of any one of claims 15-19, wherein the antisense strand comprises the base sequence of SEQ ID NO: 3, or a variant thereof.21 . The composition of any one of claims 19-20, wherein the variant comprises about 1 , or about 2, or about 3, or about 4, or about 5 mutations, the mutations selected from substitutions, deletions, and insertions.

22. The composition of any one of claims 15-19, wherein the sense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof.

23. The composition of any one of claims 15-20, wherein the antisense strand consists of the base sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or a variant thereof.

24. The composition of any one of claims 15-23, wherein the composition comprises a modified sequence selected from a nucleic acid complex comprising: a sense strand selected from5' mC#mA#mG.mA.mC.mC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 17);5' mC#mA#mG.mG.mA.mA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 18);5' mT#mA#mT.mT.mC.mT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 19);5' mC#mA#mG.mA.fC.fC.mA.mA.mA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 20);5' mC#mA#mG.mG.fA.fA.mA.mG.mA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 21);5' mT#mA#mT.mT.fC.fT.mT.mG.mG.mT.mT.mA.mA#mT#mA-Chol 3' (SEQ ID NO: 22);5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.mA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 23);5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.mT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 24);5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.mT.mA.mA#mT#niA-Chol 3' (SEQ ID NO: 25);5' mC#mA#mG.mA.fC.fC.mA.mA.fA.mG.fA.mA.mA#mG#mA-Chol 3' (SEQ ID NO: 26);5' mC#mA#mG.mG.fA.fA.mA.mG.fA.mC.fT.mG.mA#mT#mA-Chol 3' (SEQ ID NO: 27);5' mT#mA#mT.mT.fC.fT.mT.mG.fG.mT.fT.mA.mA#niT#mA-Chol 3' (SEQ ID NO: 28);5' mC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#mA-Chol 3' (SEQ ID NO: 29);5' mC#mA#fG.mG.fA.mA.fA.mG.fA.mC.fT.mG.fA#mT#mA-Chol 3' (SEQ ID NO: 30);5' mT#mA#n".mT.fC.mT.fT.mG.fG.mT.fT.mA.fA#mT#niA-Chol 3' (SEQ ID NO: 31);5' fC#mA#fG.mA.fC.mC.fA.mA.fA.mG.fA.mA.fA#mG#fA-Chol 3' (SEQ ID NO: 32);5' fC#mA#fG.mG.fA.mA.fA.mG.fA.mC.1T.mG.fA#mT#fA-Chol 3' (SEQ ID NO: 33);5' fr#mA#fT.mT.fC.mT.fr.mG.fG.mT.fr.mA.fA#mT#fA-Chol 3' (SEQ ID NO: 34); and an antisense strand of:5' PmT#fC#mT.mT.mT.mC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#nnC 3' (SEQ ID NO: 35);5' mT#fA#mT.mC.mA.mG.mT.mC.mT.mT.mT.mC.mC.1T#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 36);5'PmT#fA#mT.mT.mA.mA.mC.mC.mA.mA.mG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 37);5' PmT#fC#mT.mT.mT.fC.mT.mT.mT.rnG.mG.mT.mC.tT#mG#mC#mA#rnT#mT#mC 3' (SEQ ID NO: 38);5' PmT#fA#mT.mC.mA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#nnG#mA#mG 3' (SEQ ID NO: 39);5'PmT#fA#mT.mT.mA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 40);5'PmT#fC#mT.mT.mT.mC.mT.mT.mT.fG.fG.mT.mC.fr#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 41);5' PmT#fA#mT.mC.mA.mG.mT.mC.mT.1T.fr.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 42);5' PmT#fA#mT.mT.mA.mA.mC.mC.mA.fA.fG.mA.mA.1T#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 43);5' PmT#fC#mT.fT.fT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 44);5' PmT#fA#mT.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:45);5' PmT#fA#mT.1T.fA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:46);5' PmT#fC#fT.fT.fT.fC.mT.mT.mT.mG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO:47);5' PmT#fA#fr.fC.fA.fG.mT.mC.mT.mT.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:48);5' PmT#fA#fT.fT.fA.fA.mC.mC.mA.mA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO:49);5' PmT#fC#mT.fT.fT.fC.fT.fT.mT.fG.mG.mT.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 50);5' PmT#fA#mT.fC.fA.fG.fT.fC.mT.1T.mT.mC.mC.fT#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO: 51);5' PmT#fA#mT.1T.fA.fA.fC.fC.mA.fA.mG.mA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 52);5' PmT#fC#mT.fT.mT.fC.mT.1T.mT.fG.mG.fr.mC.fT#mG#mC#mA#mT#mT#mC 3' (SEQ ID NO: 53);5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fr.mT.fC.mC.fr#mG#mG#mT#mG#mA#mG 3' (SEQ ID NO:54);5' PmT#fA#mT.fT.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#mC#mT#mG#mA#mA 3' (SEQ ID NO: 55);5' PmT#fC#mT.fr.mT.fC.mT.fT.mT.fG.mG.fT.mC.fT#mG#fC#mA#fr#mT#fC 3' (SEQ ID NO: 56);5' PmT#fA#mT.fC.mA.fG.mT.fC.mT.fT.mT.fC.mC.fT#mG#fG#mT#fG#mA#fG 3' (SEQ ID NO: 57);5' PmT#fA#mT.fr.mA.fA.mC.fC.mA.fA.mG.fA.mA.fT#mA#fC#mT#fG#mA#fA 3' (SEQ ID NO: 58); wherein P represents 5’ phosphate, # represents a phosphorothioate linker, f represents 2' Fluoro, m represents 2' O-methyl, and Choi 3' represents Cholesterol conjugate.

25. A pharmaceutical composition comprising the composition of any one of claims 1-24, and a pharmaceutically acceptable carrier.

26. A method of promoting or enhancing degradation of an mRNA encoding USP10, comprising:administering to a subject in need thereof an effective amount of the composition of claim 1 , or pharmaceutical composition of claim 25, or contacting a cell with the composition of claim 1 , or pharmaceutical composition of claim 25, and administering to a subject in need thereof an effective amount of the composition, wherein the promoting or enhancing is as compared to a state before or without composition.

27. A method of treating a disease or disorder, comprising: administering to a subject in need thereof an effective amount of the composition of claim 1 , or pharmaceutical composition of claim 25, or contacting a cell with the composition of claim 1, or pharmaceutical composition of claim 25, and administering to a subject in need thereof an effective amount of the composition.

28. The method of claim 27, wherein the disease or disorder is characterized by scarring.

29. The method of claim 28, wherein the disease or disorder is selected from ocular scarring (optionally selected from scarring of the cornea and / or retina), integumentary scarring, internal organ scarring, and internal organ (optionally selected from lung, liver, and kidney) fibrosis.

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