CNS-Targeted Complexes and Their Use

Abstract

This application relates to a complex comprising a central nervous system (CNS) targeting agent covalently linked to a molecular payload (such as an oligonucleotide, polypeptide, small molecule, or gene therapy payload) for delivery to cells (e.g., cells of the CNS), and its use, particularly for the treatment of diseases.

Description

Technical Field

[0001] Related Applications This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63 / 367,814, filed July 6, 2022, entitled "BRAIN TARGETING COMPLEXES AND USES THEREOF", and U.S. Provisional Application Serial No. 63 / 496,184, filed April 14, 2023, entitled "CNS TARGETING COMPLEXES AND USES THEREOF", the entire contents of each of which are incorporated herein by reference.

[0002] This application relates to targeting complexes for delivering a molecular payload across the blood-brain barrier and / or to cells of the central nervous system (CNS), formulations comprising such complexes, and uses thereof, particularly for the treatment of diseases.

[0003] Reference to Electronic Sequence Listing The contents of the electronic sequence listing (D082470081WO00-SEQ-COB.xml; size: 3,883,598 bytes; created on July 5, 2023) are incorporated herein by reference in their entirety.

Background Art

[0004] Neurological diseases and disorders that affect the central nervous system (CNS) afflict millions of people worldwide, and many have few or no treatment options. Due to their limited in vivo distribution, including the inability to efficiently cross the blood-brain barrier, therapeutic compounds that have potential efficacy in various neurological conditions often fail to achieve their intended effects or have limited effectiveness.

Summary of the Invention

[0005] According to some aspects, the present disclosure provides a complex comprising a central nervous system (CNS) targeting agent covalently linked to a molecular payload, a composition comprising such a complex, and methods of using them. The CNS targeting agent of the complexes described herein has been demonstrated to be able to transport the molecular payload across the blood-brain barrier (e.g., via receptor-mediated transcytosis) and effect delivery of the molecular payload to cells of the CNS. The CNS targeting agent of the complexes described herein comprises an anti-transferrin receptor 1 (TfR1) antibody that can transport the molecular payload across the blood-brain barrier (e.g., via receptor-mediated transcytosis) and effect delivery of the molecular payload to cells of the CNS. In some embodiments, the molecular payload of the complexes described herein modulates the expression or activity of a gene associated with a central nervous system (CNS) disease or disorder and / or has a therapeutic effect on a CNS disease or disorder.

[0006] According to some aspects, a complex is provided herein, where the complex comprises an anti-TfR1 antibody covalently linked to a molecular payload for treating a central nervous system (CNS) disease or disorder, where the anti-TfR1 antibody (i) heavy chain complementarity determining region 1 (CDR-H1) of SEQ ID NO: 1, heavy chain complementarity determining region 2 (CDR-H2) of SEQ ID NO: 2, heavy chain complementarity determining region 3 (CDR-H3) of SEQ ID NO: 3, light chain complementarity determining region 1 (CDR-L1) of SEQ ID NO: 4, light chain complementarity determining region 2 (CDR-L2) of SEQ ID NO: 5, and light chain complementarity determining region 3 (CDR-L3) of SEQ ID NO: 6; (ii) CDR-H1 of SEQ ID NO: 7, CDR-H2 of SEQ ID NO: 8, CDR-H3 of SEQ ID NO: 9, CDR-L1 of SEQ ID NO: 10, CDR-L2 of SEQ ID NO: 11, and CDR-L3 of SEQ ID NO: 6; or (iii) CDR-H1 of SEQ ID NO: 12, CDR-H2 of SEQ ID NO: 13, CDR-H3 of SEQ ID NO: 14, CDR-L1 of SEQ ID NO: 15, CDR-L2 of SEQ ID NO: 5, and CDR-L3 of SEQ ID NO: 16; where the complex delivers the molecular payload to cells of the CNS and comprises.

[0007] In some embodiments, the anti-TfR1 antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 18.

[0008] In some embodiments, the anti-TfR1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 19 and a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0009] In some embodiments, the anti-TfR1 antibody is a Fab.

[0010] In some embodiments, the molecular payload is configured to regulate the expression of a gene associated with a CNS disease or disorder.

[0011] In some embodiments, the molecular payload comprises an oligonucleotide, a polypeptide, a small molecule, or a gene therapy payload. In some embodiments, the gene therapy payload comprises a messenger RNA (mRNA) molecule.

[0012] In some embodiments, the anti-TfR1 antibody is covalently linked to the molecular payload via a linker having the structure of formula (I):

Chemical Structure

[0013] In some embodiments, the complex comprises a structure of formula (J):

Chemical formula

[0014] In some embodiments, the complex delivers the molecular payload to cells of the CNS by passing through the blood-brain barrier. In some embodiments, the complex delivers the molecular payload to cells of the CNS by passing through the choroid plexus.

[0015] In some embodiments, the gene associated with a CNS disease or disorder is DMPK, DMD, SMN, or FXN.

[0016] In some embodiments, the gene associated with a CNS disease or disorder is SOD1, C9orf72, ATXN2, or FUS.

[0017] In some embodiments, the gene associated with a CNS disease or disorder is LRRK2 or SNCA.

[0018] In some embodiments, the gene associated with a CNS disease or disorder is HTT or MSH3.

[0019] In some embodiments, the gene associated with a CNS disease or disorder is TREM2, APOE, MAPT, or APP.

[0020] In some embodiments, the gene associated with a CNS disease or disorder is GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, or SCN9A.

[0021] In some embodiments, the gene associated with a CNS disease or disorder is SCN1A, SCN2A, SCN8A, SCN9A, CLN3, GRIA1, or PCDH19.

[0022] In some embodiments, the gene associated with a CNS disease or disorder is TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1.

[0023] In some embodiments, the gene associated with a CNS disease or disorder is PIKFYVE, SYF2, or UNC13A.

[0024] In some embodiments, the gene associated with a CNS disease or disorder is GRIN2A.

[0025] In some embodiments, the gene associated with a CNS disease or disorder is ATXN1, ATXN2, ATXN3, or MSH3.

[0026] In some embodiments, the gene associated with a CNS disease or disorder is GRN, C9orf72, MAPT, PIKFYVE, SYF2, or UNC13A.

[0027] In some embodiments, the gene associated with a CNS disease or disorder is TPP1 or CLN3.

[0028] In some embodiments, the gene associated with a CNS disease or disorder is APOE, SCN1A, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2.

[0029] In some embodiments, the molecular payload comprises an oligonucleotide that comprises a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 392 - 702 or to a target sequence of an oligonucleotide listed in any one of Tables 5 - 19. In some embodiments, the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 - 19.

[0030] In some embodiments, the molecular payload comprises an oligonucleotide that comprises a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 705 - 803 or to a target sequence of an oligonucleotide listed in any one of Tables 5 - 19. In some embodiments, the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 - 19.

[0031] In some embodiments, the molecular payload comprises an oligonucleotide that comprises a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 143 - 148, 167 - 169, 810 - 875, and 1059 - 1068 or to a target sequence of an oligonucleotide listed in any one of Tables 5 - 19. In some embodiments, the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 - 19.

[0032] In some embodiments, the CNS disease or disorder is a neuromuscular disease or disorder. In some embodiments, the neuromuscular disease or disorder is Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy.

[0033] In some embodiments, the CNS disease or disorder is amyotrophic lateral sclerosis.

[0034] In some embodiments, the CNS disease or disorder is Parkinson's disease.

[0035] In some embodiments, the CNS disease or disorder is essential tremor.

[0036] In some embodiments, the CNS disease or disorder is Huntington's disease.

[0037] In some embodiments, the CNS disease or disorder is Alzheimer's disease.

[0038] In some embodiments, the CNS disease or disorder is hereditary dystonia.

[0039] In some embodiments, the CNS disease or disorder is epilepsy.

[0040] In some embodiments, the CNS disease or disorder is a pain disorder.

[0041] In some embodiments, the CNS disease or disorder is glycogen synthesis disorder; neurodegeneration; small fiber neuropathy; phenotypes related to nociception; Alexander disease; Angelman syndrome; autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis.

[0042] In some embodiments, the CNS disease or disorder is spinocerebellar ataxia (SCA).

[0043] In some embodiments, the CNS disease or disorder is frontotemporal dementia (FTD).

[0044] In some embodiments, the CNS disease or disorder is a motor neuron disease.

[0045] In some embodiments, the CNS disease or disorder is Dravet syndrome.

[0046] In some embodiments, the CNS disease or disorder is Batten disease.

[0047] In some embodiments, the CNS disease or disorder is GM1 gangliosidosis.

[0048] In some embodiments, the CNS disease or disorder is Niemann-Pick type A.

[0049] In some embodiments, the CNS disease or disorder is metachromatic leukodystrophy.

[0050] In some embodiments, the CNS disease or disorder is Krabbe disease.

[0051] In some embodiments, the CNS disease or disorder is Tay-Sachs.

[0052] In some embodiments, the CNS disease or disorder is Sandhoff disease.

[0053] In some embodiments, the CNS disease or disorder is Gaucher disease type II or III.

[0054] In some embodiments, the CNS disease or disorder is Rett syndrome.

[0055] In some embodiments, the CNS disease or disorder is such that the molecular payload is the molecular payload disclosed in any one of paragraphs 0216 - 1208. In some embodiments, the molecular payload is the molecular payload disclosed in any one of paragraphs 0296 - 0299, 0404 - 0406, 0468 - 0470, 0500 - 0502, 0535 - 0539, 0601 - 0604, 0666 - 0668, 0757 - 0759, 0779 - 0781, 0896 - 0901, 0916 - 0918, 0946 - 0948, 1049 - 1056, 1070 - 1078, 1092 - 1102, 1116 - 1124, 1138 - 1141, 1155 - 1158, 1172 - 1177, and 1191 - 1193.

[0056] According to some aspects, a method of treating a CNS disease or disorder is provided herein, where the method comprises administering to a subject in need thereof a complex as disclosed herein.

[0057] According to some aspects, a method of delivering a molecular payload to the CNS of a subject is provided herein, where the method comprises administering to the subject a complex as disclosed herein.

[0058] In some embodiments, the complex is administered intravenously to the subject.

[0059] In some embodiments, the complex is detectable in the cortex of the subject after administration.

[0060] In some embodiments, the complex is detectable in the cerebellum of the subject after administration.

[0061] In some embodiments, the complex is detectable in the deep brain tissue of the subject after administration. In some embodiments, the deep brain tissue is the thalamus, caudate nucleus, and / or putamen of the subject.

[0062] In some embodiments, the complex is detectable in the cortical neurons, motor neurons, cerebellar cells, and / or choroid plexus cells of the subject after administration.

[0063] In some embodiments, the molecular payload comprises a protein. In some embodiments, the protein is an enzyme.

[0064] In some embodiments, the subject is diagnosed with or suspected of having Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, or Gaucher disease.

[0065] In some embodiments, the payload comprises an oligonucleotide.

[0066] In some embodiments, the subject is diagnosed with or suspected of having ALS, Angelman syndrome, Rett syndrome, Parkinson's, Lewy body dementia, Alzheimer's disease (which may or may not be associated with cerebral amyloid angiopathy (CAA) or frontotemporal dementia), epilepsy, Alexander disease, spinal muscular atrophy, Batten disease, Huntington's disease, spinocerebellar ataxia, motor neuron disease, or Dravet syndrome. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0079] Aspects of the present disclosure relate to the recognition that, due to limitations in their in vivo distribution, which include that certain molecular payloads (e.g., oligonucleotides, peptides, small molecules, gene therapy) can have beneficial effects in cells of the central nervous system (CNS) but cannot efficiently cross the blood-brain barrier, it has proven challenging for such molecular payloads to achieve their intended effects in the CNS. Accordingly, the present disclosure, in some aspects, provides a complex comprising a CNS targeting agent covalently linked to a molecular payload to overcome such challenges. The CNS targeting agents of the present disclosure include anti-transferrin receptor 1 (TfR1) antibodies that have been demonstrated to be capable of transporting a molecular payload to cells of the CNS. In some embodiments, such delivery occurs across the blood-brain barrier (e.g., via receptor-mediated transcytosis) and results in delivery of the molecular payload to cells of the CNS. In some embodiments, such delivery occurs through the choroid plexus and results in delivery of the molecular payload to cells of the CNS. In some embodiments, the anti-TfR1 antibodies of the complexes described herein exhibit pH-dependent binding affinity for TfR1 (e.g., having different binding affinities under different pH conditions). In some embodiments, the anti-TfR1 antibodies of the complexes described herein exhibit pH-independent binding affinity for TfR1 (e.g., having comparable binding affinities under different pH conditions).

[0080] In some embodiments, the complexes provided herein can include a molecular payload that regulates (e.g., increases or decreases) the expression and / or activity of genes associated with CNS diseases and disorders by modulating transcription, translation, post-transcriptional modifications (such as splicing), mRNA stability, and / or protein stability. In some embodiments, the complexes provided herein can include a molecular payload that is a synthetic nucleic acid (such as DNA or RNA) that can be used to express one or more proteins that regulate the expression and activity of genes associated with CNS diseases and disorders. In some embodiments, the complexes provided herein can include a molecular payload that has a therapeutic effect in a CNS disease or disorder, but may or may not modulate the expression or activity of any gene associated with CNS diseases and disorders.

[0081] Neurological diseases and disorders that affect the central nervous system (CNS) have diverse etiologies and potential treatment modalities. Examples of such CNS diseases and disorders include, without limitation, in particular, neuromuscular disorders (such as myotonic dystrophy, Duchenne muscular dystrophy, Friedreich's ataxia, and spinal muscular atrophy), amyotrophic lateral sclerosis (ALS), frontotemporal dementia (FTD), Parkinson's disease, Huntington's disease, Alzheimer's disease, epilepsy, and pain disorders. Other examples of CNS diseases and disorders include essential tremor and hereditary dystonia. Additional examples of CNS diseases and disorders include spinocerebellar ataxia, motor neuron diseases, Dravet syndrome, Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, Gaucher disease types II and III, and Rett syndrome. Various genes are involved in CNS diseases and disorders, including, without limitation, in particular, DMPK, DMD, SMN, FXN, SOD1, C9orf72, ATXN2, FUS, LRRK2, SNCA, HTT, MSH3, TREM2, APOE, MAPT, APP, GYS1, PrP, VLA-4, UBE3A, GFAP, LSD, SCN9A, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, and PCDH19. Other genes involved in CNS diseases and disorders include, without limitation, TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, and ECHS1. Additional genes involved in CNS diseases and disorders include, without limitation, PIKFYVE, SYF2, UNC13A, ATXN1, ATXN3, GRN, GRIN2A, TPP1, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, and MECP2. Molecular payloads can be effective in treating CNS diseases and disorders by their delivery to cells of the CNS, for example, by passing through the blood-brain barrier and / or by passing through the choroid plexus.Certain molecular payloads can alleviate the signs or symptoms of CNS diseases and disorders, for example, in some embodiments, by modulating the expression or activity of genes involved in CNS diseases and disorders. Molecular payloads that alleviate the signs or symptoms of CNS diseases and disorders without modulating the expression or activity of any gene involved in CNS diseases and disorders can also be used according to the present disclosure. Delivery of a molecular payload to the CNS can also be useful for other purposes other than the treatment of CNS diseases and disorders.

[0082] Further aspects of the disclosure, including explanations of defined terms, are provided below. I. Definitions

[0083] Administer: As used herein, the term “administer” or “administration” means to provide a complex to a subject in a manner that is physiologically and / or (by way of example, and) pharmacologically useful (e.g., to treat a condition in the subject).

[0084] Alzheimer's disease: As used herein, the term "Alzheimer's disease" refers to a progressive neurological disorder characterized by atrophy of brain tissue and loss of neurons, particularly due to old age. Alzheimer's disease is a frequent cause of dementia, including mild cognitive impairment. The symptoms of Alzheimer's disease include memory loss that worsens over time, difficulty concentrating, particularly on abstract concepts, difficulty multitasking, impaired decision-making, and changes in personality or behavior. Alzheimer's disease is also associated with the formation of beta-amyloid protein plaques and tau protein tangles (also known as neurofibrillary changes) in brain tissue. These are cytotoxic, block cell communication, and contribute to neuron death. The cause of Alzheimer's disease is incompletely understood. However, the development of Alzheimer's disease can be influenced by the inheritance of certain genetic risk factors. For example, genes involved in the pathophysiology of Alzheimer's disease include, but are not limited to, TREM2, APOE, MAPT, and APP (see, for example, Neuner SM, et al. “Genetic architecture of Alzheimer’s disease.” Neurobiol Dis. 2020;143:104976; and Ibanez L, et al. “Advances in Genetic and Molecular Understanding of Alzheimer’s Disease.” Genes (Basel). 2021; 12(8):1247). Alzheimer's disease may or may not be associated with cerebral amyloid angiopathy (CAA) or frontotemporal dementia.

[0085] Amyotrophic Lateral Sclerosis (ALS): Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that affects nerve cells in the central nervous system. ALS causes the degeneration of motor neurons that control muscle movement, ultimately leading to the loss of control of the muscles required for movement, speech, eating, and breathing. Approximately 90% of cases of ALS are considered sporadic and occur in patients without a known family history of the disease, and 5-10% of all cases are familial (i.e., hereditary). Genes associated with the development of ALS include, for example, SOD1 (associated with approximately 12-20% of cases of familial ALS), C9orf72 (associated with approximately 25-40% of cases of familial ALS), ATXN2, and FUS. In some aspects, the accumulation of TDP-43 aggregates is associated with ALS. PIKFYVE, SYF2, and UNC13A are also involved in the pathophysiology of ALS. In some aspects, single nucleotide polymorphism(s) and / or other modulation(s) in PIKFYVE, SYF2, and UNC13A are associated with ALS.

[0086] ANO3: As used herein, ANO3 refers to the gene encoding a protein belonging to the TMEM16 family of predicted membrane proteins, anoctamin 3 (also known as DYT23; DYT24; TMEM16C; C11orf25; or GENX-3947). In some embodiments, ANO3 can be a human (Gene ID: 63982), non-human primate (e.g., Gene ID: 101865236), or rodent gene (e.g., Gene ID: 228432, Gene ID: 311287). In humans, mutations in the gene encoding ANO3 are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_001313726.2; NM_031418.4; XM_047427399.1; XM_017018118.3; NM_001313727.2; XM_017018119.3; and XM_011520282.4) have been characterized.

[0087] Antibody: As used herein, the term "antibody" refers to a polypeptide that includes at least one immunoglobulin variable domain or at least one antigen determinant, for example, a paratope that specifically binds to an antigen. In some embodiments, the antibody is a full-length antibody. In some embodiments, the antibody is a chimeric antibody. In some embodiments, the antibody is a humanized antibody. However, in some embodiments, the antibody is a Fab fragment, Fab' fragment, F(ab')2 fragment, Fv fragment, or scFv fragment. In some embodiments, the antibody is a nanobody derived from a camelid antibody or a nanobody derived from a shark antibody. In some embodiments, the antibody is a diabody. In some embodiments, the antibody includes a framework having human germline sequences. In another embodiment, the antibody includes a heavy chain constant domain selected from the group consisting of IgG, IgG1, IgG2, IgG2A, IgG2B, IgG2C, IgG3, IgG4, IgA1, IgA2, IgD, IgM, and IgE constant domains. In some embodiments, the antibody includes a heavy (H) chain variable region (abbreviated as VH herein) and / or (for example, and) a light (L) chain variable region (abbreviated as VL herein). In some embodiments, the antibody includes a constant domain, for example, an Fc region. The immunoglobulin constant domain refers to a heavy or light chain constant domain. The amino acid sequences of human IgG heavy and light chain constant domains and their functional variants are known. For the heavy chain, in some embodiments, the heavy chain of the antibody described herein can be an alpha (α), delta (δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In some embodiments, the heavy chain of the antibody described herein can include a human alpha (α), delta (δ), epsilon (ε), gamma (γ), or mu (μ) heavy chain. In certain embodiments, the antibody described herein includes a human gamma1 CH1, CH2, and / or (for example, and) CH3 domain. In some embodiments, the amino acid sequence of the VH domain includes the amino acid sequence of a human gamma (γ) heavy chain constant region such as any known in the art.Non-limiting examples of human constant region sequences are described in the art. See, for example, U.S. Pat. No. 5,693,780 and Kabat E A et al., (1991) supra. In some embodiments, the VH domain comprises an amino acid sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or at least 99% identical to any of the variable chain constant regions provided herein. In some embodiments, the antibody is modified. For example, it is modified via glycosylation, phosphorylation, SUMOylation, and / or (by way of example, and) methylation. In some embodiments, the antibody is a glycosylated antibody conjugated to one or more sugar or carbohydrate molecules. In some embodiments, the one or more sugar or carbohydrate molecules are conjugated to the antibody via N-glycosylation, O-glycosylation, C-glycosylation, GPI-anchoring (attachment of a GPI anchor), and / or (by way of example, and) phosphoglycosylation. In some embodiments, the one or more sugar or carbohydrate molecules are monosaccharides, disaccharides, oligosaccharides, or glycans. In some embodiments, the one or more sugar or carbohydrate molecules are branched oligosaccharides or branched glycans. In some embodiments, the one or more sugar or carbohydrate molecules include mannose units, glucose units, N-acetylglucosamine units, N-acetylgalactosamine units, galactose units, fucose units, or lipid phosphate units. In some embodiments, the antibody is a construct comprising a polypeptide that includes a linker polypeptide or one or more antigen-binding fragments of the present disclosure linked to an immunoglobulin constant domain. The linker polypeptide includes two or more amino acid residues joined by peptide bonds and is used to join one or more antigen-binding moieties. Examples of linker polypeptides have been reported (see, for example, Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, R. J., et al. (1994) Structure 2:1121-1123).Further, the antibody can be part of a larger immune adhesion molecule formed by covalent or non-covalent binding of the antibody or antibody portion to one or more other proteins or peptides. Examples of such immune adhesion molecules include the use of streptavidin core regions to create tetrameric scFv molecules (Kipriyanov, S. M., et al. (1995) Human Antibodies and Hybridomas 6:93-101) and the use of cysteine residues, marker peptides, and C-terminal polyhistidine tags to create bivalent and biotinylated scFv molecules (Kipriyanov, S. M., et al. (1994) Mol. Immunol. 31:1047-1058).

[0088] APP: As used herein, the term "APP" refers to the gene encoding the protein involved in synapse formation and neuroplasticity, amyloid beta precursor protein (also referred to as AAA, ABETA, ABPP, AD1, APPI, CTF gamma, CVAP, PN-II, PN2, alpha-sAPP, and preA4). In some embodiments, APP can be a human (Gene ID: 351), non-human primate (e.g., Gene ID: 100427716), or rodent gene (e.g., Gene ID: 11820, Gene ID: 54226). In humans, mutations in the APP gene are associated with the development of Alzheimer's disease. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000484.4 and NM_201413.3) have been characterized.

[0089] APOE: As used herein, APOE refers to the gene encoding the protein apolipoprotein E (also referred to as AD2, ApoE4, LDLCQ5, and LPG), which is involved in the formation of lipoprotein particles and the transport of lipids through the circulatory system. In some embodiments, APOE can be a human (Gene ID: 348), non-human primate (e.g., Gene ID: 714623), or rodent gene (e.g., Gene ID: 11816, Gene ID: 25728). In humans, mutations in the gene encoding APOE are associated with the development of Alzheimer's disease. In some embodiments, the APOE4 allele is associated with the development of Alzheimer's disease. In some embodiments, the APOE4 allele is associated with the development of motor neuron disease. Thus, in some embodiments, allele-specific regulation (e.g., suppression) of APOE (e.g., APOE4) is useful for the treatment of CNS diseases or disorders such as Alzheimer's disease or motor neuron disease. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_000041.4 and NM_001302688.2) have been characterized.

[0090] Approximately: As used herein, the term "approximately" or "about" when applied to one or more values of interest refers to a value that is similar to the stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values that fall within 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the stated reference value in both (greater or lesser) directions, unless otherwise stated or otherwise apparent from the context (except where such number exceeds 100% of the possible value).

[0091] ARSA: As used herein, ARSA refers to the gene encoding arylsulfatase A (cerebroside-sulfatase, epididymal secretory sperm-binding protein, MLD, ASA, or sulfatidase also), an enzyme that degrades sulfide. In some embodiments, ARSA can be a human (e.g., gene ID: 410), non-human primate (e.g., gene ID: 458946, gene ID: 716500), or rodent (e.g., gene ID: 11883, gene ID: 315222) gene. In humans, mutations in the gene encoding ARSA are associated with the development of metachromatic leukodystrophy. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_000487.6; NM_001085425.3; NM_001085426.3; NM_001085427.3; NM_001085428.3; NM_001362782.2; XM_047441363.1; XM_024452241.2; XM_011530691.4) have been characterized.

[0092] ASM: As used herein, the term "ASM" refers to the gene encoding acid sphingomyelinase (also known as SCMPD1, ASMASE, and NPD). Wild-type acid sphingomyelinase is a lysosomal enzyme involved in the conversion of lipids to ceramides. Mutations in ASM typically result in a deficiency or complete loss of function of acid sphingomyelinase, leading to the accumulation of fat in cells of various organs and tissues, including the central nervous system. In some embodiments, ASM can be a human gene (gene ID: 6609), non-human primate gene (gene ID: 711248), or rodent gene (gene ID: 20597; gene ID: 308909). In humans, mutations in ASM are associated with the development of Niemann-Pick type A.

[0093] ATP1A3: As used herein, ATP1A3 refers to a protein belonging to the family of P-type cation-transporting ATPases and the family of Na+ / K+ ATPases, and to the gene encoding ATPase Na+ / K+ transporting subunit alpha 3 (also referred to as RDP; AHC2; CAPOS; DEE99; DYT12; or ATP1A1). In some embodiments, ATP1A3 can be a human (gene ID: 478), non-human primate (e.g., gene ID: 102122869), or rodent gene (e.g., gene ID: 232975, gene ID: 24213). In humans, mutations in the gene encoding ATP1A3 are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_152296.5; NM_001256213.2; NM_001256214.2; and XM_047438862.1) have been characterized.

[0094] ATXN1: As used herein, ATXN1 refers to the gene encoding the protein ataxin-1. The ataxin-1 protein is expressed throughout the body and is thought to be involved in controlling protein production, including transcription and RNA processing. Ataxin-1 binds to RNA and associates with large protein complexes. It is thought to be involved in transcriptional repression and to regulate developmental processes controlled by Notch and Capicua. Human ATXN1 (Gene ID: 6310) contains a CAG repeat region, which normally contains 6 to 39 repeats. Longer expansions of the CAG repeat region in ATXN1 (typically 40 to 83 or more) can lead to neurodegenerative diseases, including spinocerebellar ataxia type 1 (SCA1). The expanded CAG repeat region results in incorrect protein folding and non-functional ataxin-1 protein. The abnormal protein forms aggregates in the cell nucleus and causes cell damage. Evidence suggests that ataxin-1 aggregates are found primarily or mainly in cells of the CNS, particularly within Purkinje cells of the cerebellum. The accumulation of protein aggregates results in cell death; the loss of these cells over time leads to the cerebellar deficits characteristic of SCA1. See Banfi, et al. “Identification and characterization of the gene causing type 1 spinocerebellar ataxia” Nature Genet. 7:513-520 (1994), and Orr, et al. “Expansion of an unstable trinucleotide CAG repeat in spinocerebellar ataxia type 1” Nature Genet. 4: 221-226 (1993). Allele-specific inhibition of ATXN1 may be effective in the treatment of SCA1. See Miller, et al. “Allele-specific silencing of dominant disease genes” Proc. Nat. Acad. Sci. 100: 7195-7200 (2003).

[0095] ATXN2: As used herein, ATXN2 refers to the gene encoding the protein ataxin-2. It is ubiquitously expressed in various tissues, and the ataxin-2 protein localizes to the Golgi apparatus and stress granules in normal cells. The ataxin-2 protein is involved in controlling mRNA translation through its interaction with poly(A)-binding protein and is also involved in the formation of stress granules and P bodies. Both of these are also involved in RNA regulation. Human ATXN2 contains a CAG repeat region. This normally contains 22 or 23 repeats, but can contain up to 31 repeats. Longer expansions of the CAG repeat region in ATXN2 can lead to neurodegenerative diseases including spinocerebellar ataxia type 2 (SCA2) and amyotrophic lateral sclerosis (ALS). The interaction between ataxin-2 and the protein TDP-43 is thought to be involved in the development of ALS in certain patients. See, for example, Elden, et al. “Ataxin-2 intermediate-length polyglutamine expansions are associated with increased risk for ALS” Nature 466:1069-1075 (2010). Disease-associated ATXN2 alleles often contain 34 to 52 CAG repeats, but can contain as few as 32 or more than 100, and the size can expand when the allele is transmitted to subsequent generations. In some embodiments, as described by Elden, et al., as few as 27 CAG repeats can be associated with ALS. Allele-specific inhibition of ATXN2 may be effective in the treatment of SCA2. See Miller, et al. “Allele-specific silencing of dominant disease genes” Proc. Nat. Acad. Sci. 100: 7195-7200 (2003).

[0096] ATXN3: As used herein, ATXN3 refers to the gene encoding the ataxin-3 protein (also known as AT3, ATX3, JOS, MJD, MJD1, and SCA3). The ataxin-3 protein is expressed throughout the body and is believed to be involved in the proteasome processing system. Ataxin-3 removes ubiquitin from proteins to be degraded so that ubiquitin can be recycled. Ataxin-3 may also be involved in controlling the first stage of transcription. Human ATXN2 (Gene ID: 4287) contains a CAG repeat region, which normally contains 13 to 36 repeats. Longer expansions of the CAG repeat region in ATXN3 (typically 50 or more) can lead to neurodegenerative diseases including spinocerebellar ataxia type 3 (SCA3). The expanded CAG repeat region results in incorrect protein folding and non-functional ataxin-3 protein. This non-functional ataxin-3 protein cannot remove ubiquitin from proteins and leads to the aggregation of such proteins together with ubiquitin and ataxin-3 in the cell nucleus. These protein aggregates can lead to cell death including neurons and other cells in the CNS. Neurons are typically the cell type most affected by mutations in ATXN3. See Kawaguchi, et al. “CAG expansions in a novel gene for Machado-Joseph disease at chromosome 14q32.1” Nature Genet. 8: 221-228 (1994). Allele-specific inhibition of ATXN3 may be effective in the treatment of SCA3. See Miller, et al. “Allele-specific silencing of dominant disease genes” Proc. Nat. Acad. Sci. 100: 7195-7200 (2003).

[0097] Batten disease: As used herein, the term "Batten disease" refers to a family of lysosomal disorders also known as neuronal ceroid lipofuscinosis (NCL). Batten disease is a neurological disorder caused by various mutations in 13 genes that typically inherit in an autosomal recessive pattern. In some embodiments, Batten disease results from mutations in CLN1, CLN2, CLN3, CLN4, CLN5, CLN6, CLN7, CLN8, CLN10, CLN11, CLN12, CLN13, or CLN14. In particular, in some embodiments relevant to the present disclosure, Batten disease results from mutations in CLN2 or CLN3. The symptoms of Batten disease include seizures, visual impairment, reduced cognitive and behavioral function, reduced motor function, developmental disorders, and early death. Batten disease is characterized by lysosomal accumulation of autofluorescent storage material, glial reactivity, and neuronal loss. The genetic causes of Batten disease are well-known and can be attributed to mutations in one of 13 different genes encoding lysosomal and extra-lysosomal proteins. The genes involved in the pathophysiology of Batten disease include PPT1 (CLN1), TPP1 (CLN2), CLN3 (CLN3), DNAJC5 (CLN4), CLN5 (CLN5), CLN6 (CLN6), MFSD8 (CLN7), CLN8 (CLN8), CTSD (CLN10), GRN (CLN11), ATP13A2 (CLN12), CTSF (CLN13), and KCTD7 (CLN14). In some embodiments, a subject in need of treatment for Batten disease exhibits seizure activity. In some embodiments, seizure activity includes myoclonic jerks, grand mal seizures, and tonic-clonic seizures. In some embodiments, a subject in need of treatment for Batten disease exhibits symptoms of visual impairment. In some embodiments, visual impairment includes optic atrophy, progressive loss of vision, retinitis pigmentosa, macular degeneration, visual field defects, retinopathy, reduced light reflex, loss of central vision, and blindness. In some embodiments, a subject in need of treatment for Batten disease exhibits symptoms of reduced cognitive and behavioral function. In some embodiments, reduced cognitive and behavioral function includes irritability, hyperexcitability, anxiety, agitation, depression, inappropriate laughter, mood disorders, intellectual disability, dementia, and personality disorders.In some embodiments, a subject in need of treatment for Batten disease exhibits symptoms of motor function decline. In some embodiments, motor function decline includes loss of motor coordination, choreoathetosis, stereotyped movements, myoclonic ataxia, hypokinesia, spasticity, dystonic features, hypotonia, muscle contracture, balance disorder, myoclonus, ataxia, facial dyskinesia, clumsiness, loss of motor coordination, dysarthria, severe respiratory distress, central, axial, and / or limb hypotonia, limb spasticity, tremors, parkinsonism, hyperreflexia, aphasia, echolalia, delayed speech, and dysarthric speech. In some embodiments, a subject in need of treatment for Batten disease exhibits symptoms of developmental disorder. In some embodiments, developmental disorder includes decelerated head growth, early death, microcephaly, suture riding, interruption of developmental milestones, developmental regression, and developmental arrest.

[0098] Blood-brain barrier: As used herein, the term "blood-brain barrier" refers to a highly selective semipermeable boundary of endothelial cells that prevents various molecules in the blood from non-selectively passing into the extracellular fluid of the CNS. It allows the passage of some small molecules by diffusion and the active transport of various nutrients, ions, organic anions, and macromolecules such as glucose and amino acids that are essential for nerve function, but blocks the non-specific transport of other molecules.

[0099] C9orf72: As used herein, C9orf72 refers to the gene encoding the chromosome 9 open reading frame 72 protein. The protein is found in many regions of the brain, including the cytoplasm of neurons and presynaptic terminals. Mutations that cause disease in the C9orf72 gene, particularly hexanucleotide repeat expansions, are associated with ALS. In some embodiments, mutations in C9orf72 are associated with the familial form of ALS. In some embodiments, mutations in C9orf72 are associated with frontotemporal dementia (such as C9FTD).

[0100] CDR: As used herein, the term "CDR" refers to the complementarity-determining regions within an antibody variable sequence. A typical antibody molecule includes a heavy-chain variable region (VH) and a light-chain variable region (VL), which are usually involved in antigen binding. The VH and VL regions can be further subdivided into regions of hypervariability, also known as "complementary-determining regions" ("CDRs"), which are interspersed with more conserved regions known as "framework regions" ("FRs"). Each VH and VL typically consists of three CDRs and four FRs arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The extent of the framework regions and CDRs can be accurately identified using methodologies known in the art, for example, by the Kabat definition, the IMGT definition, the Chothia definition, the AbM definition, and / or (by way of example, and) the contact definition. All of these are well-known in the art.For example, see Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242; IMGT (R), the international ImMunoGeneTics information system (R) imgt.org, Lefranc, M.-P. et al., Nucleic Acids Res., 27:209-212 (1999); Ruiz, M. et al., Nucleic Acids Res., 28:219-221 (2000); Lefranc, M.-P., Nucleic Acids Res., 29:207-209 (2001); Lefranc, M.-P., Nucleic Acids Res., 31:307-310 (2003); Lefranc, M.-P. et al., In Silico Biol., 5, 0006 (2004) [Epub], 5:45-60 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 33:D593-597 (2005); Lefranc, M.-P. et al., Nucleic Acids Res., 37:D1006-1012 (2009); Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015); Chothia et al., (1989) Nature 342:877; Chothia, C. et al. (1987) J. Mol. Biol. 196:901-917, Al-lazikani et al (1997) J. Molec. Biol. 273:927-948; and Almagro, J. Mol. Recognit. 17:132-143 (2004). See also hgmp.mrc.ac.uk and bioinf.org.uk / abs.As used herein, CDR can refer to a CDR defined by any method known in the art. Two antibodies having the same CDR means that the two antibodies have the same amino acid sequence of that CDR determined by the same method, e.g., the IMGT definition.

[0101] Each of the variable regions of the heavy and light chains has three CDRs, which are designated CDR1, CDR2, and CDR3 for each of the variable regions. As used herein, the term "CDR set" refers to the group of three CDRs present on a single variable region capable of binding to an antigen. The exact boundaries of these CDRs are defined differently according to different systems. The system described by Kabat (Kabat et al., Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987) and (1991)) not only provides an unambiguous residue numbering system applicable to any variable region of an antibody, but also provides the exact residue boundaries that define the three CDRs. These CDRs can be referred to as Kabat CDRs. The subparts of the CDRs can be referred to as L1, L2, and L3, or H1, H2, and H3, where "L" and "H" refer to the light and heavy chain regions, respectively. These regions can be referred to as Chothia CDRs, which have boundaries that overlap with the Kabat CDRs. Other boundaries that define CDRs overlapping with the Kabat CDRs are described by Padlan (FASEB J. 9:133-139 (1995)) and MacCallum (J Mol Biol 262(5):732-45 (1996)). Still other definitions of CDR boundaries may not strictly follow one of the above systems, but will still overlap with the Kabat CDRs. However, they can be shorter or longer in light of predictions or experimental findings that a particular residue or group of residues or even an entire CDR has no significant impact on antigen binding. The methods used herein can utilize CDRs defined according to any of these systems. An example of a CDR definition system is provided in Table 1.

Table 1

[0102] CDR-grafted antibody: The term "CDR-grafted antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species, but in which one or more of the CDR region sequences of VH and / or (by way of example, and) VL have been replaced by CDR sequences from another species. By way of example, an antibody having mouse heavy and light chain variable regions in which one or more (e.g., CDR3) of the mouse CDRs have been replaced by human CDR sequences.

[0103] Central nervous system (CNS): As used herein, the term "central nervous system" (CNS) refers to the brain and spinal cord and includes neurons and non-neuronal supporting cells (e.g., glia) as well as blood-brain barrier cells. The blood-brain barrier prevents the non-selective passage of various molecules from the circulation into the extracellular fluid of the CNS. The CNS also includes cells of the blood-cerebrospinal fluid barrier, such as the cells of the choroid plexus.

[0104] CNS disease or disorder: As used herein, "CNS disease or disorder" refers to a disease or disorder that affects and / or has an etiology in the CNS (e.g., the structure or function of the CNS). CNS diseases and disorders are also known as neurological diseases or disorders. Examples of CNS diseases or disorders include, but are not limited to, neuromuscular diseases and disorders (e.g., muscular dystrophy, myotonic dystrophy, spinal muscular atrophy, and Friedreich's ataxia), amyotrophic lateral sclerosis (ALS), Parkinson's disease, Huntington's disease, Alzheimer's disease, epilepsy, and pain disorders. Other examples of CNS diseases or disorders include essential tremor and hereditary dystonia. Certain lysosomal storage disorders are also examples of CNS diseases or disorders.

[0105] CNS targeting agent: As used herein, the term "CNS targeting agent" refers to a molecule that specifically binds to an antigen expressed on cells of the CNS (e.g., neurons, supporting cells, and / or cells of the blood-brain barrier). Antigens within or on CNS cells can be membrane proteins, such as integral membrane proteins or surface membrane proteins. Typically, a CNS targeting agent specifically binds to an antigen on CNS cells, which facilitates the transport of a molecular payload across the blood-brain barrier and / or the internalization of the CNS targeting agent (and any associated molecular payload) into CNS cells. In some embodiments, the CNS targeting agent can specifically bind to an internalizing cell surface receptor (e.g., transferrin receptor 1) on cells of the CNS and be internalized into CNS cells through receptor-mediated internalization. In some embodiments, the CNS targeting agent is a small molecule, protein, peptide, nucleic acid (e.g., aptamer), or antibody. In some embodiments, the CNS targeting agent is linked to a molecular payload.

[0106] CNS targeting antibody: As used herein, the term "CNS targeting antibody" refers to a CNS targeting agent that is an antibody that specifically binds to an antigen found within or on CNS cells. In some embodiments, the CNS targeting antibody specifically binds to an antigen on CNS cells (e.g., neurons, supporting cells, and / or cells of the blood-brain barrier), which facilitates the transport of a molecular payload across the blood-brain barrier and / or the internalization of the CNS targeting antibody (and any associated molecular payment) into CNS cells. In some embodiments, the CNS targeting antibody specifically binds to an internalizing cell surface receptor present on CNS cells. In some embodiments, the CNS targeting antibody facilitates transcytosis across cells of the blood-brain barrier (e.g., endothelial cells). In some embodiments, the CNS targeting antibody is an antibody that specifically binds to a transferrin receptor (e.g., transferrin receptor 1).

[0107] Chimeric antibody: The term "chimeric antibody" refers to an antibody that contains heavy and light chain variable region sequences from one species and constant region sequences from another species, for example, an antibody having mouse heavy and light chain variable regions linked to human constant regions.

[0108] CLN2: See "TPP1".

[0109] CLN3: As used herein, the term "CLN3" refers to the gene encoding the CLN3 lysosomal / endosomal transmembrane protein (also referred to as battenin, BTN1, BTS, and JNCL), a protein involved in lysosomal function. In some embodiments, CLN3 can be a human (gene ID: 1201), non-human primate (e.g., gene ID: 705815), or rodent gene (e.g., gene ID: 12752, gene ID: 293485). In humans, mutations in the gene encoding CLN3 are associated with epilepsy and seizures as well as CLN3 Batten disease. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000086.2 and NM_001286104.2) have been characterized.

[0110] Complementary: As used herein, the term "complementary" refers to the ability of two nucleosides or two sets of nucleosides to form exact pairs. In particular, complementary is a term that characterizes the degree of hydrogen bond pair formation that results in a bond between two nucleosides or two sets of nucleosides. For example, if the base at one position of an oligonucleotide can hydrogen bond with the base at the corresponding position of a target nucleic acid (e.g., mRNA), the bases are considered complementary to each other at that position. Base pair formation can include both classical Watson-Crick base pair formation and non-Watson-Crick base pair formation (e.g., Wobble base pair formation and Hoogsteen base pair formation). For example, in some embodiments, for complementary base pair formation, an adenosine-type base (A) is complementary to a thymidine-type base (T) or a uracil-type base (U), a cytosine-type base (C) is complementary to a guanosine-type base (G), and universal bases such as 3-nitropyrrole or 5-nitroindole can hybridize to any A, C, U, or T and are considered complementary to it. Inosine (I) is also considered a universal base in the art and is considered complementary to any A, C, U, or T.

[0111] Conservative Amino Acid Substitutions: As used herein, "conservative amino acid substitutions" refer to amino acid substitutions that do not modulate the relative charge or size characteristics of the protein in which the amino acid substitution is made. Variants can be prepared according to methods known to those of skill in the art for modulating polypeptide sequences. By way of example, references compiling such methods can be found, for example, in Molecular Cloning: A Laboratory Manual, J. Sambrook, et al., eds., Fourth Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2012, or Current Protocols in Molecular Biology, F.M. Ausubel, et al., eds., John Wiley & Sons, Inc., New York. Conservative substitutions of amino acids include substitutions made between amino acids within the following groups: (a) M, I, L, V; (b) F, Y, W; (c) K, R, H; (d) A, G; (e) S, T; (f) Q, N; and (g) E, D.

[0112] Covalently Linked: As used herein, the term "covalently linked" refers to the characteristic that two or more molecules are linked together through at least one covalent bond. In some embodiments, two molecules can be covalently linked together by a single bond serving as an intermolecular linker, such as a disulfide bond or disulfide bridge. However, in some embodiments, two or more molecules can be covalently joined together through a molecule serving as a linker that joins two or more molecules together through multiple covalent bonds. In some embodiments, the linker can be a cleavable linker. However, in some embodiments, the linker can be a non-cleavable linker.

[0113] Cross-reactivity: The term "cross-reactivity" as used herein and in the context of targeting agents (e.g., CNS targeting agents such as antibodies) refers to the property of an agent that can specifically bind to more than one antigen of a similar type or class (e.g., antigens of multiple homologs, paralogs, or orthologs) with similar affinity or avidity. For example, in some embodiments, an antibody that is cross-reactive with human and non-human primate antigens of a similar type or class (e.g., human transferrin receptor and non-human primate transferrin receptor) can bind to human and non-human primate antigens with similar affinity or avidity. In some embodiments, the antibody is cross-reactive with human antigens and rodent antigens of a similar type or class. In some embodiments, the antibody is cross-reactive with rodent antigens and non-human primate antigens of a similar type or class. In some embodiments, the antibody is cross-reactive with human antigens, non-human primate antigens, and rodent antigens of a similar type or class.

[0114] DMD: When used herein, the term "DMD" refers to the gene that encodes the dystrophin protein, a key component of the dystrophin-glycoprotein complex that cross-links the internal cytoskeleton and extracellular matrix in muscle cells, particularly muscle fibers. Deletions, duplications, and point mutations in DMD can cause dystrophinopaties such as Duchenne muscular dystrophy, Becker muscular dystrophy, or cardiomyopathy. Alternative promoter usage and alternative splicing result in numerous distinct transcript variants and protein isoforms of this gene. In some embodiments, the dystrophin gene can be a human (Gene ID: 1756), non-human primate (e.g., Gene ID: 465559), or rodent gene (e.g., Gene ID: 13405; Gene ID: 24907). In addition, multiple human transcript variants encoding different protein isoforms (as annotated by, for example, GenBank RefSeq accession numbers: NM_000109.3, NM_004006.2, NM_004009.3, NM_004010.3, and NM_004011.3) have been characterized.

[0115] DMD allele: As used herein, the term "DMD allele" refers to any one of alternative forms (e.g., wild-type or mutant forms) of the DMD gene. In some embodiments, a DMD allele can encode dystrophin that retains its normal and typical function. In some embodiments, a DMD allele can contain one or more mutations that result in muscular dystrophy. Common mutations leading to Duchenne muscular dystrophy involve frameshift, deletion, substitution, and duplication mutations in one or more of the 79 exons present in the dystrophin allele, such as exons 8, 23, 41, 44, 45, 50, 51, 52, 53, or 55. Further examples of DMD mutations are disclosed, for example, in Flanigan KM, et al., Mutational spectrum of DMD mutations in dystrophinopathy patients: application of modern diagnostic techniques to a large cohort. Hum Mutat. 2009 Dec; 30 (12):1657-66, the contents of which are hereby incorporated by reference in their entirety.

[0116] DMPK: As used herein, the term "DMPK" refers to the gene encoding the serine / threonine protein kinase myotonic protein kinase (also known as myotonic dystrophy protein kinase or dystrophia myotonica protein kinase). Substrates for this enzyme can include myogenin, the beta subunit of the L-type calcium channel, and phospholemman. In some embodiments, DMPK can be a human (Gene ID: 1760), non-human primate (e.g., Gene ID: 456139, Gene ID: 715328), or rodent gene (e.g., Gene ID: 13400). In humans, CTG repeat expansions in the 3' non-coding untranslated region of DMPK are associated with myotonic dystrophy type I (DM1). Additionally, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_001081563.2, NM_004409.4, NM_001081560.2, NM_001081562.2, NM_001288764.1, NM_001288765.1, and NM_001288766.1) have been characterized.

[0117] DMPK Allele: As used herein, the term "DMPK allele" refers to any one of alternative forms of the DMPK gene (e.g., wild-type or mutant forms). In some embodiments, a DMPK allele can encode a wild-type myotonic protein kinase that retains its normal and typical function. In some embodiments, a DMPK allele can contain a repeat expansion associated with one or more diseases. In some embodiments, a normal subject has two DMPK alleles that contain repeat units in the range of 5 to 37. In some embodiments, the number of CTG repeat units in a subject with DM1 ranges from about 50 to about 3,000 or more, and higher repeat numbers lead to increased severity of the disease. In some embodiments, a mild DM1 subject has at least one DMPK allele having repeat units in the range of 50 to 150. In some embodiments, a subject with classical DM1 has at least one DMPK allele having repeat units in the range of 100 to 1,000 or more. In some embodiments, a subject with DM1 having a congenital onset can have at least one DMPK allele containing more than 2,000 repeat units.

[0118] Dravet Syndrome: As used herein, the term "Dravet syndrome" is also known as severe myoclonic epilepsy in infancy (SMEI) and refers to the most severe disorder in the spectrum of genetic epilepsy with febrile seizures plus (GEFS+). Dravet syndrome is typically caused by de novo mutations, although cases arising from familial mutations also occur. The symptoms of Dravet syndrome include seizures, cognitive decline, developmental regression, intellectual disability, and ataxia. More than 80% of cases of Dravet syndrome are attributable to mutations in SCN1A, in which more than 900 distinct mutations have been reported. In some embodiments, a subject in need of treatment for Dravet syndrome has symptoms selected from: seizures (e.g., febrile seizures, afebrile seizures, myoclonic seizures, and absence seizures), cognitive decline, developmental regression, intellectual disability, and ataxia.

[0119] Dystrophinopathy: As used herein, the term "dystrophinopathy" refers to a muscle or neurological disorder resulting from one or more mutated DMD alleles. Dystrophinopathy encompasses a spectrum of conditions (ranging from mild to severe) that includes Duchenne muscular dystrophy, Becker muscular dystrophy, and DMD-related dilated cardiomyopathy (DCM). In some embodiments, at one end of the spectrum, dystrophinopathy is phenotypically associated with asymptomatic increases in the serum concentration of creatine phosphokinase (CK) and / or (by way of example, and) muscle cramps with myoglobinuria. In some embodiments, at the other end of the spectrum, dystrophinopathy is phenotypically associated with progressive muscle disorders. These are generally classified as Duchenne or Becker muscular dystrophy when the skeletal muscles are primarily affected and as DMD-related dilated cardiomyopathy (DCM) when the heart is primarily affected. The symptoms of Duchenne muscular dystrophy include muscle loss or degeneration, diminished muscle function, pseudohypertrophy of the tongue and calf muscles, a higher risk of neurological abnormalities, and a shortened lifespan. Duchenne muscular dystrophy is associated with the Online Mendelian Inheritance in Man (OMIM) entry #310200. Becker muscular dystrophy is associated with the OMIM entry #300376. Dilated cardiomyopathy is associated with the OMIM entry X#302045.

[0120] ECHS1: As used herein, ECHS1 refers to the gene encoding enoyl-CoA hydratase short chain 1 (SCEH; mECH; mECH1; or also referred to as ECHS1D), a protein that functions in the second step of the mitochondrial fatty acid beta-oxidation pathway. In some embodiments, ECHS1 can be a human (Gene ID: 1892), non-human primate (e.g., Gene ID: 101925228), or rodent gene (e.g., Gene ID: 93747, Gene ID: 140547). In humans, mutations in the gene encoding ECHS1 are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, as in GenBank RefSeq accession number NM_004092.4) have been characterized.

[0121] Epilepsy: As used herein, the term "epilepsy" refers to a neurological disorder characterized by abnormal activity in the neurons of the brain, which can cause intermittent seizures and may be accompanied by loss of consciousness. The symptoms of an epileptic seizure can include sudden confusion or anxiety, dizziness, loss of awareness, loss of consciousness, staring, muscle rigidity, and involuntary movements of the limbs. Epileptic seizures occur frequently in the absence of external stimuli. Epileptic seizures can occur due to abnormal neurological activity in a single brain region (focal seizures) or due to abnormal neurological activity throughout the brain (generalized seizures). Focal seizures can occur with or without loss of consciousness and can cause modified muscle movement and / or sensory perception. Generalized seizures can further be characterized as absence seizures (i.e., petit mal seizures that occur briefly and cause loss of awareness and repetitive body movements), myoclonic seizures (sudden involuntary muscle movements), tonic seizures (muscle rigidity), atonic seizures (loss of muscle control, without rigidity), and tonic-clonic seizures (i.e., grand mal seizures that cause loss of consciousness, muscle rigidity, and sudden involuntary muscle movements). Epilepsy can cause death due to persistent injury during seizures, status epilepticus, or sudden unexpected death in epilepsy (SUDEP). Only approximately half of epilepsy cases have identifiable causes such as developmental disorders, brain abnormalities (such as brain tumors, traumatic brain injury, or vascular disorders such as stroke), brain-infecting infectious diseases, or inheritance of genetic risk factors.For example, genes involved in the pathophysiology of epilepsy include, but are not limited to, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, and PCDH19 (see, for example, Wang J, et al. “Epilepsy-associated genes.” Seizure. 2017; 44:11-20; Abdennadher M, et al. “Seizure phenotype in CLN3 disease and its relation to other neurologic outcome measures.” J Inherit Metab Dis. 2021; 44(4):1013-1020; and Samanta D “PCDH19-Related Epilepsy Syndrome: A Comprehensive Clinical Review.” Pediatr Neurol. 2020; 105:3-9). GRIN2A is also involved in the pathophysiology of epilepsy in some aspects.

[0122] Essential tremor: As used herein, the term "essential tremor" refers to a neurological condition characterized by involuntary shaking movements. It is also known in some cases as familial tremor or benign essential tremor. Essential tremor affects both men and women and is most common in people over the age of 40. The tremor is most likely to be observed in the forearm and hand, and the upper arm, head, eyelids, and other muscles can also be affected. Persons with essential tremor may have problems gripping or using small objects such as silverware or writing utensils. The tremors associated with essential tremor most commonly involve small, rapid movements that occur 4 to 12 times per second. Specific symptoms can include head nodding, tremors, or a vibrating voice (if the tremor affects the larynx), and problems with writing, drawing, drinking from a cup, or using tools (for example, if the tremor affects the hand and / or forearm). Essential tremor typically worsens over time and can be severe in some patients. The exact cause of essential tremor is unknown, but many cases of essential tremor have a genetic origin and are inherited in an autosomal dominant pattern. Mutated genes associated with essential tremor can affect various regions of the brain, including deep brain regions such as the thalamus and the cerebellum.

[0123] Framework: As used herein, the term "framework" or "framework array" refers to the remaining array of variable regions minus the CDRs. Since the exact definition of the CDR arrays can be determined by different systems, the meaning of the framework array correspondingly receives different interpretations. Also, the six CDRs (CDR-L1, CDR-L2, and CDR-L3 of the light chain, and CDR-H1, CDR-H2, and CDR-H3 of the heavy chain) divide the framework regions on the light and heavy chains into four subregions (FR1, FR2, FR3, and FR4) on each chain. Herein, CDR1 is located between FR1 and FR2, CDR2 is located between FR2 and FR3, and CDR3 is located between FR3 and FR4. Without specifying a particular subregion as FR1, FR2, FR3, or FR4, the framework region otherwise referred to represents the combined FRs within the variable region of a single naturally occurring immunoglobulin chain. As used herein, FR (singular) represents one of the four subregions, and FRs (plural) represents two or more of the four subregions that make up the framework region. Human heavy and light chain acceptor sequences are known in the art. In one aspect, acceptor sequences known in the art can be used in the antibodies disclosed herein.

[0124] Friedreich's ataxia: As used herein, the term "Friedreich's ataxia" refers to an autosomal recessive genetic disorder caused by mutations in the FXN gene and characterized by progressive damage to muscle tissue and the nervous system. Friedreich's ataxia is a neurological disorder associated with an expansion of GAA trinucleotide repeats in the FXN gene that leads to a decrease in FXN expression. The expanded GAA trinucleotide repeats located within the first intron form R-loops, which can interfere with normal transcription processes and reduce FXN gene expression. The FXN allele in healthy individuals contains <36 GAA repeats, whereas in FRDA patients, the GAA expansion, which ranges from 70 to 1700 GAA repeats, leads to FXN mRNA deficiency and subsequently reduced levels of frataxin, a mitochondrial protein encoded by a nuclear gene essential for life (see, for example, Silva et al., “Expanded GAA repeats impair FXN gene expression and reposition the FXN locus to the nuclear lamina in single cells.” Hum. Molec. Genet., 2015, Vol. 24, No. 12 3457-3471).Friedreich's ataxia, the genetic basis of the disease, and related symptoms are described in the art (see, for example, Montermini, L. et al. “The Friedreich’s ataxia GAA triplet repeat: premutation and normal alleles.” Hum. Molec. Genet., 1997, 6: 1261-1266.; Filla, A. et al. “The relationship between trinucleotide (GAA) repeat length and clinical features in Friedreich’s ataxia.” Am. J. Hum. Genet. 1996, 59: 554-560.; Pandolfo, M. Friedreich’s ataxia: the clinical picture. J. Neurol. 2009, 256, 3-8). Friedreich's ataxia is associated with the Online Mendelian Inheritance in Man (OMIM) entry #229300.

[0125] Frontotemporal dementia: As used herein, the term "frontotemporal dementia" or "FTD" refers to a disease with progressive degeneration of the frontal and / or temporal lobes of the brain. FTD results in progressive deficits in behavior, executive function, and / or language, and the symptoms include changes in social and personal behavior, apathy, emotional blunting, lack of expressive language, and lack of language processing. FTD is considered one of the most prevalent forms of dementia, corresponding to 10% - 20% of all dementia cases. FTD can generally be categorized into (i) behavioral variant FTD (bVFTD), (ii) primary progressive aphasia (PPA), (iii) progressive supranuclear palsy (PSP), and (iv) corticobasal syndrome (CBS). Many cases of FTD are linked to mutations occurring in C9orf72, granulin (GRN), and MAPT. Additionally, pathologically, there are three major protein deposits found in the brains of FTD patients, TAR DNA-binding protein 43 (TDP-43), Fused in sarcoma (FUS), and tau. In some embodiments, a subject in need of treatment for FTD has a mutation in the GRN gene, the C9orf72 gene, and / or the MAPT gene. In some embodiments, a subject in need of treatment for FTD has progressive degeneration of the frontal and / or temporal lobes of the brain. In some embodiments, a subject in need of treatment for FTD has TAR DNA-binding protein 43 (TDP-43), Fused in sarcoma (FUS), and / or tau deposits in the brain. In some embodiments, a subject in need of treatment for FTD has deficits in behavior, executive function, and / or language. In some embodiments, a subject in need of treatment for FTD has one or more of the following symptoms: changes in social and personal behavior, apathy, emotional blunting, lack of expressive language, and lack of language processing.

[0126] FUS: As used herein, FUS refers to the gene encoding the RNA-binding protein FUS / TLS, also known as heterogeneous nuclear ribonucleoprotein P2. This protein is a subunit of a complex involved in pre-mRNA maturation and has also been shown to be involved in the DNA repair response. Loss of function of the protein encoded by FUS results in increased DNA damage in neurons, and certain mutations in FUS impair the PARP-dependent DNA damage response, leading to neurodegeneration and the formation of RNA-binding protein FUS / TSL aggregates. Some mutations in FUS have been identified in ALS patients. See, for example, Kwiatkowski, et al., “Mutations in the FUS / TLS Gene on Chromosome 16 Cause Familial Amyotrophic Lateral Sclerosis” Science 323(5918):1205-1205 (2009) and Vance, et al., “Mutations in FUS, an RNA Processing Protein, Cause Familial Amyotrophic Lateral Sclerosis Type 6” Science 323(5918):1208-1211 (2009). The mechanism by which FUS mutations cause ALS is not known. However, it is believed that toxicity likely results from a gain of toxic cytoplasmic function, since many FUS mutations linked to ALS are located on its nuclear localization signal and mouse models that do not express FUS, and thus have a complete loss of FUS nuclear localization, do not develop overt ALS-like symptoms.

[0127] FXN: As used herein, the term "FXN" refers to the gene encoding the protein frataxin, which is involved in iron homeostasis. In some embodiments, FXN can be a human (Gene ID: 2395), non-human primate (e.g., Gene ID: 737660), or rodent gene (e.g., Gene ID: 14297, Gene ID: 499335). In humans, a GAA repeat expansion in the first intron of FXN is associated with the neurological disorder Friedreich's ataxia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated by GenBank RefSeq accession numbers: NM_000144.4 and NM_181425.2, for example) have been characterized.

[0128] GALC: As used herein, "GALC" refers to the gene encoding the lysosomal protein galactosylceramidase (also referred to as GALC and entrez: 2581). Galactosylceramidase degrades galactolipids involved in myelin production. In some embodiments, GALC can be a human (e.g., Gene ID: 2581), non-human primate (e.g., Gene ID: 693322, Gene ID: 736519), or rodent (e.g., Gene ID: 14420, Gene ID: 314360). In humans, mutations in the GALC gene are associated with the development of Krabbe disease. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated by GenBank RefSeq accession numbers: NM_000153.4; NM_001201401.2; NM_001201402.2; XM_011536618.3; XM_047431198.1; XM_047431199.1, for example) have been characterized.

[0129] Gaucher disease types II and III: As used herein, the terms "Gaucher disease" or "GD" refer to Gaucher disease types II and III, which are genetic disorders in which fatty substances (such as glucocerebroside) accumulate in cells and certain organs (such as the spleen and liver). The accumulation of these fatty substances causes the organs to enlarge and can negatively affect organ function. When the bones are affected, it can weaken the bones, and when the bone marrow is affected, it can interfere with clotting. Type II Gaucher disease is a form of Gaucher disease that affects the central nervous system, spleen, liver, lungs, and bones. Type II Gaucher disease (also known as Gaucher type II and acute infantile neuronal Gaucher disease) presents symptoms within the first year of life. Symptoms of Gaucher type II include developmental delay, abnormal eye movements, hypertonia, laryngeal spasm, seizures, chronic lung infections, splenomegaly, and hepatomegaly. Current enzyme replacement therapies for Gaucher type I and Gaucher type III are not effective in Gaucher type II. Gaucher type II is a fatal disease that usually results in death within the first two years of life. Gaucher type III (also known as chronic neuronal Gaucher disease) occurs in childhood. Initial symptoms of Gaucher type III are hepatosplenomegaly, poor appetite, and less weight gain than normal. Other symptoms include seizures, skeletal disturbances, abnormal eye movements, cognitive problems, poor coordination, respiratory problems, and blood disorders. Both Gaucher type II and type III are neuropathic. In addition, both are associated with mutations in the GBA gene. In some embodiments, a subject in need of treatment for Gaucher disease has a mutation in the GBA gene. In some embodiments, a subject in need of treatment for Gaucher disease has glucocerebroside accumulation in cells and / or organs. In some embodiments, a subject in need of treatment for Gaucher disease has hepatomegaly and / or splenomegaly. In some embodiments, a subject in need of treatment for Gaucher disease has one or more of the following symptoms: abnormal eye movements, hypertonia, laryngeal spasm, seizures, chronic lung infections, splenomegaly, hepatomegaly. In some embodiments, a subject in need of treatment for Gaucher disease has one or more of the following symptoms: poor appetite, less weight gain than normal, skeletal disturbances, abnormal eye movements, cognitive problems, poor coordination, respiratory problems, and blood disorders.

[0130] GBA: As used herein, the term "GBA" refers to the gene (also referred to as GBA1, GCB, GLUC) that encodes the lysosomal membrane protein β-glucocerebrosidase (also known as acid β-glucosidase, D-glucosyl-N-acylsphingosine glucohydrolase, glucosylceramidase beta, glucocerebrosidase, or GC-ase). β-glucocerebrosidase cleaves β-glucoside linkages. In some embodiments, GBA can be human (e.g., gene ID: 2629), non-human primate (e.g., gene ID: 719103), or rodent (e.g., gene ID: 14466, gene ID: 684536). In humans, mutations in the GBA gene are associated with the development of Gaucher disease types II and III. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000157.4; NM_001005741.3; NM_001005742.3; NM_001171811.2; NM_001171812.2) have been characterized.

[0131] GCH1: As used herein, GCH1 refers to the gene encoding a protein that is a member of the GTP cyclohydrolase family and is the first and rate-limiting enzyme in tetrahydrobiopterin (BH4) biosynthesis, which catalyzes the conversion of GTP to 7,8-dihydroneopterin triphosphate, also known as guanosine triphosphate cyclohydrolase 1 (GCH; DYT5; DYT14; DYT5a; GTPCH1; HPABH4B; GTP-CH-1). In some embodiments, GCH1 can be a human (Gene ID: 2643), non-human primate (e.g., Gene ID: 695675), or rodent gene (e.g., Gene ID: 14528, Gene ID: 29244). In humans, mutations in the gene encoding GCH1 are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_001024071.2; NM_001024070.2; NM_001024024.2; NM_000161.3; XM_017021218.2; and XM_047431261.1) have been characterized.

[0132] GFAP: As used herein, the term "GFAP" refers to the gene encoding the glial fibrillary acidic protein (also known as ALXDRD), a protein involved in cell-cell communication in the CNS. In some embodiments, GFAP can be a human (Gene ID: 2670), non-human primate (e.g., Gene ID: 712941), or rodent gene (e.g., Gene ID: 14580, Gene ID: 24387). In humans, mutations in the GFAP gene are associated with Alexander disease. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_002055.5 and NM_001131019.3) have been characterized.

[0133] GLB1: As used herein, the term "GLB1" refers to the gene encoding beta-galactosidase 1, a lysosomal enzyme that mediates the catabolism of several molecules including GM1 ganglioside (also known as EBP, ELNR1, or MPS4B). GM1 ganglioside is an important factor in neuronal plasticity, neuronal repair, and the release of neurotrophins in the brain. When mutated, GLB1 produces beta-galactosidase 1 with reduced or lost function, leading to the accumulation of GM1 ganglioside in the brain and ultimately resulting in neuronal death. GLB1 mutations are associated with GM1 gangliosidosis and Morquio syndrome type B. In some embodiments, GLB1 can be a human gene (Gene ID: 2720), a non-human primate gene (Gene ID: 709355), or a rodent gene (Gene ID: 12091; Gene ID: 316033).

[0134] GM1 gangliosidosis: As used herein, the term "GM1 gangliosidosis" refers to a lysosomal storage disorder caused by a deficiency of the β-galactosidase enzyme. GM1 gangliosidosis is a neurological disorder that is inherited in an autosomal recessive pattern and is associated with mutations in the GLB1 gene. The symptoms of GM1 gangliosidosis include cognitive impairment, developmental delay, skeletal abnormalities, seizures, movement disorders, and visual impairment. GM1 gangliosidosis is characterized by neuronal cell death and demyelination, inflammatory responses, autophagy, and mitochondrial dysfunction. The genetic basis of GM1 gangliosidosis has been attributed to mutations in GLB1, and there are 102 reported mutations of this. See Brunetti-Pierri, et al. “GM1 gangliosidosis: review of clinical, molecular, and therapeutic aspects” Mol Gen Metabolism 94(4): 391-396 (2008). GM1 gangliosidosis is closely related to both Tay-Sachs and Sandhoff diseases; thus, treatments for Tay-Sachs and / or Sandhoff diseases may also be effective in treating GM1 gangliosidosis (and vice versa).

[0135] GNAL: As used herein, GNAL refers to the protein, a stimulatory G protein alpha subunit that mediates odorant signal transduction in the olfactory epithelium, and the gene encoding the G protein subunit alpha L (also referred to as HG1O and DYT25). The G protein subunit alpha L protein couples the dopamine type 1 receptor and the adenosine A2A receptor and is widely expressed in the central nervous system. In some embodiments, GNAL can be a human (Gene ID: 2774), non-human primate (e.g., Gene ID: 102137826), or rodent gene (e.g., Gene ID: 14680, Gene ID: 24611). In humans, mutations in the gene encoding GNAL are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_182978.4; NM_001142339.3; NM_001261443.2; NM_001261444.2; NM_001369387.1; and XM_006722324.4) have been characterized.

[0136] GRIA1: As used herein, the term "GRIA1" refers to the gene encoding the protein involved in neuronal signaling via the glutamic acid neurotransmitter, the AMPA-type subunit 1 of the ionotropic glutamate receptor (also referred to as GLUH1, GLUR1, GLURA, GluA1, and HBGR1). In some embodiments, GRIA1 can be a human (Gene ID: 2890), non-human primate (e.g., Gene ID: 714117), or rodent gene (e.g., Gene ID: 14799, Gene ID: 50592). In humans, mutations in the GRIA1 gene are associated with epilepsy and seizures, as well as phenotypes related to nociception (e.g., pain disorders). In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000827.4 and NM_001114183.2) have been characterized.

[0137] GRIN2A: As used herein, the term GRIN2A refers to the gene encoding the protein, glutamate ionotropic receptor NMDA type subunit 2A (gluN2A), which is one component of a subset of NMDA receptors (also known as LKS; EPND; FESD; NR2A; GluN2A; NMDAR2A). In some embodiments, GRIN2A can be a human (Gene ID: 2903), non-human primate (e.g., Gene ID: 102123126), or rodent gene (e.g., Gene ID: 14811, Gene ID: 24409). In humans, mutations in the GRIN2A gene are associated with epilepsy and seizures. Over 50 mutations in GRIN2A have been identified in patients with epilepsy. Many GRIN2A mutations lead to the production of non-functional gluN2A proteins or prevent the production of gluN2A proteins, presumably leading to a reduction in the number of functional NMDA receptors. Thus, signaling occurs more often through other types of NMDA receptors that are more easily stimulated, resulting in excessive signaling in the brain. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000833.5; NM_001134407.3; NM_001134408.2) have been characterized.

[0138] GRN: As used herein, GRN refers to the gene encoding the protein progranulin (also known as granulin precursor, proepithelin, and PC cell-derived growth factor) that is active in many tissues throughout the body (also known as GEP; GP88; PEPI; PGRN; CLN11; PCDGF). The function of progranulin in the brain is not well understood, but it appears to play an important role in neuron survival. In humans, mutations in GRN are associated with frontotemporal dementia (FTD). FTD associated with GRN mutations (sometimes also referred to as "GRN-FTD" or "GRN-related FTD") has been suggested to involve the accumulation of TAR DNA-binding protein 43 into aggregates in certain cells of the central nervous system, including neurons in the brain. These protein aggregates can interfere with cell function and lead to cell death.For example, see Baker, et al. “Mutations in progranulin cause tau-negative frontotemporal dementia linked to chromosome 17” Nature 442: 916-919 (2006); Cruts, et al., “Null mutations in progranulin cause ubiquitin-positive frontotemporal dementia linked to chromosome 17q21” Nature 442: 920-924 (2006); Borroni, et al. “Progranulin genetic variations in frontotemporal lobar degeneration: evidence for low mutation frequency in an Italian clinical series” Neurogenetics 9: 197-205 (2008); and Chen-Plotkin, et al. “Genetic and clinical features of progranulin-associated frontotemporal lobar degeneration” Arch.Neurol. 68: 488-497 (2011); the entire contents of each of these are hereby incorporated by reference into this specification. See also Hsiung, et al. “GRN Frontotemporal Dementia” 2007 Sep 7 [Updated 2020 Feb 6]. In: Adam MP, et al., editors. GeneReviews (R) [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2023. Available from: ncbi.nlm.nih.gov / books / NBK1371 / . Mutations in progranulin that cause deletions can play a role in lysosomal dysfunction.In some embodiments, the GRN can be human (e.g., Gene ID: 2896), non-human primate (e.g., Gene ID: 454728, Gene ID: 714851), or rodent (e.g., Gene ID: 14824, Gene ID: 29143). In humans, mutations in the gene encoding GRN are associated with the development of FTD, e.g., heterozygous mutations that cause insufficient production of progranulin. A human transcript variant annotated with GenBank RefSeq accession number NM_002087.4 encoding progranulin has been characterized.

[0139] GYS1: As used herein, the term "GYS1" refers to the gene encoding glycogen synthase, a protein that functions in glycogen synthesis. In some embodiments, GYS1 can be human (Gene ID: 2997), non-human primate (e.g., Gene ID: 574233, Gene ID: 456196, Gene ID: 102134439), or rodent gene (e.g., Gene ID: 14936, Gene ID: 690987). In humans, the expression of mutant glycogen synthase protein (e.g., from a mutant GYS1 gene) results in decreased glycogen synthesis. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_001161587.1 and NM_002103.5) have been characterized.

[0140] Hereditary Dystonia: As used herein, the term "hereditary dystonia" refers to a movement disorder characterized by abnormal, often repetitive movements and / or postures, resulting from sustained or intermittent muscle contractions. Dystonic movements are typically patterned and torsional and may be associated with tremors. Some forms of hereditary dystonia are associated with neurodegeneration and deteriorate progressively over time, while other forms are independent of neurodegeneration and typically reach a plateau after an initial period of deterioration. Hereditary dystonia can be characterized by the body part(s) affected and is typically classified as focal, affecting one body part (e.g., eyelids, mouth, larynx, neck, or hands and arms); segmental, affecting two or more contiguous body parts (e.g., axial - neck and trunk; brachial - one arm and trunk or both arms + / - neck + / - trunk; or crural - one leg and trunk or both legs + / - trunk); multifocal, affecting two or more non - contiguous body parts (e.g., cranio - brachial - blepharospasm and hand / arm); unilateral, affecting two or more body parts (e.g., ipsilateral arm and leg); or generalized, affecting three or more body parts (e.g., trunk and two or more other sites, + / - involvement of legs). Numerous genes associated with hereditary dystonia include TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, and ECHS1. The hereditary dystonia, the genetic basis of the disease, and the related symptoms are described in the art (see, e.g., Klein, et al., “Hereditary Dystonia Overview” 2003 Oct 28 (Updated 2017 Jun 22) In: Adam, et al., editors, GeneReviews [Internet], Seattle (WA): University of Washington, Seattle, 1993 - 2023, NCBI Bookshelf ID: NBK1155, PMID 20301334).

[0141] HEXA: As used herein, "HEXA" refers to the gene (also referred to as TSD and hexosaminidase subunit alpha) that encodes the alpha subunit of the enzyme beta - hexosaminidase A (also referred to as hexosaminidase A), an enzyme that degrades molecules containing GM2 ganglioside and N - acetylhexosamine. Mutations in HEXA reduce or eliminate the activity of beta - hexosaminidase A, resulting in the accumulation of GM2 ganglioside in neuronal cells, which can lead to cell death. In some embodiments, HEXA can be from a human (e.g., gene ID: 3073), a non - human primate (e.g., gene ID: 698251, gene ID: 748732), or a rodent (e.g., gene ID: 15211, gene ID: 300757). In humans, mutations in the gene encoding HEXA are associated with the development of Tay - Sachs. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_000520.6; NM_001318825.2) have been characterized.

[0142] HEXB: As used herein, "HEXB" refers to the gene (also referred to as ENC-1AS, HEL-248, and HEL-S-111) that encodes the β subunit of the enzyme β-hexosaminidase A (also referred to as hexosaminidase A). Wild-type HEXB is a subunit of β-hexosaminidase A that is involved in the breakdown of ganglioside GM2 and other molecules. Mutations in HEXB reduce or abolish the activity of β-hexosaminidase A, resulting in the accumulation of GM2 ganglioside in neuronal cells, which can lead to cell death. In some embodiments, HEXB can be a human gene (Gene ID: 3074), a non-human primate gene (Gene ID: 704464), or a rodent gene (Gene ID: 15212; Gene ID: 294673). In addition, multiple human transcript variants encoding different protein isoforms (such as those annotated by GenBank RefSeq accession numbers: NM_000521.4; NM_001292004.2) have been characterized. In humans, mutations in HEXB are associated with the development of Sandhoff disease.

[0143] Human antibody: As used herein, the term "human antibody" is intended to encompass antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibodies of the present disclosure can include amino acid residues not encoded by human germline immunoglobulin sequences (such as mutations introduced by in vitro random or site-directed mutagenesis or by in vivo somatic mutations), for example, on the CDRs, particularly on CDR3. However, as used herein, the term "human antibody" is not intended to encompass antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto a human framework sequence.

[0144] Humanized antibody: The term "humanized antibody" refers to an antibody that contains heavy and light chain variable region sequences from a non-human species (such as a mouse), but in which at least some portions of the VH and / or (such as, and) VL sequences have been modified to be more "human-like", i.e., more similar to human germline variable sequences. One type of humanized antibody is a CDR-grafted antibody, in which human CDR sequences are introduced onto non-human VH and VL sequences to replace the corresponding non-human CDR sequences. In one embodiment, humanized anti-TfR1 antibodies and antigen-binding portions are provided. Such antibodies can be made by obtaining a mouse anti-TfR1 monoclonal antibody using conventional hybridoma technology and then humanizing it using in vitro genetic engineering, such as that disclosed in Kasaian et al PCT Publication No. WO 2005 / 123126 A2.

[0145] HTT: As used herein, HTT refers to the gene encoding the huntingtin protein. HTT is widely expressed and required for normal development. The exact function of the huntingtin protein encoded by HTT is not known, but it plays an important role in nerve cells and is involved in axonal transport. HTT is expressed in many tissues throughout the body and has the highest expression level in the brain. Huntingtin has been found to directly interact with numerous other proteins, including several involved in transcription, transport, and cell signaling. Certain mutations in HTT result in the development of Huntington's disease. HTT contains a CAG trinucleotide repeat region. CAG trinucleotide repeat expansions are associated with Huntington's disease. In some embodiments, normal subjects have two HTT alleles containing from about 10 to about 35 CAG repeats. In some embodiments, subjects having or expected to develop Huntington's disease have an HTT allele containing 40 or more CAG repeats. In some embodiments, subjects having one or two HTT alleles containing 36 - 40 CAG repeats may or may not develop symptoms of Huntington's disease. Mutant HTT is also referred to as mHTT.

[0146] Huntington's disease: Huntington's disease is a neurological disorder characterized by the degeneration of striatal neurons. It results in the progressive degeneration of nerve cells in the brain, has a broad impact on the patient's functional abilities, and typically causes motor, cognitive, and psychiatric disorders. Huntington's disease affects the entire brain, but certain regions of the brain are more highly impacted than others. The striatum, which plays a key role in the control of movement, mood, and behavior, is usually the part of the brain most affected by Huntington's disease. Huntington's disease is associated with an expansion of the CAG trinucleotide repeat in HTT.

[0147] Internalizing cell surface receptor: As used herein, the term "internalizing cell surface receptor" refers to a cell surface receptor that is internalized by a cell, for example, by an external stimulus, such as ligand binding to the receptor. In some embodiments, the internalizing cell surface receptor is internalized by endocytosis. In some embodiments, the internalizing cell surface receptor is internalized by clathrin-mediated endocytosis. However, in some embodiments, the internalizing cell surface receptor is internalized by a clathrin-independent pathway, such as, for example, phagocytosis, macropinocytosis, uptake mediated by caveolae and rafts, or constitutive clathrin-independent endocytosis. In some embodiments, the internalizing cell surface receptor comprises an intracellular domain, a transmembrane domain, and / or (by way of example, and) an extracellular domain, which may optionally further comprise a ligand-binding domain. In some embodiments, the cell surface receptor becomes internalized by the cell after ligand binding. In some embodiments, the ligand can be a CNS targeting agent or a CNS targeting antibody. In some embodiments, the internalizing cell surface receptor is the transferrin receptor.

[0148] Isolated antibody: As used herein, "isolated antibody" is intended to refer to an antibody that is substantially free of other antibodies having different antigen specificities (for example, an isolated antibody that specifically binds to the transferrin receptor is substantially free of antibodies that specifically bind to antigens other than the transferrin receptor). However, an isolated antibody that specifically binds to a transferrin receptor complex may have cross-reactivity to other antigens, such as transferrin receptor molecules from other species. Moreover, the isolated antibody may be substantially free of other cellular materials and / or (by way of example, and) chemical substances.

[0149] Kabat numbering: The terms "Kabat numbering," "Kabat definitions," and "Kabat labeling" are used interchangeably herein. These terms, as recognized in the art, refer to a system for numbering amino acid residues that are more variable (i.e., hypervariable) than other amino acid residues in the heavy and light chain variable regions of an antibody or other antigen-binding portion thereof (Kabat et al. (1971) Ann. NY Acad. Sci. 190:382-391 and Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242). For the heavy chain variable region, the hypervariable regions are in the range of amino acid positions 31-35 for CDR1, 50-65 for CDR2, and 95-102 for CDR3. For the light chain variable region, the hypervariable regions are in the range of amino acid positions 24-34 for CDR1, 50-56 for CDR2, and 89-97 for CDR3.

[0150] KMT2B: When used herein, KMT2B refers to the gene encoding lysine methyltransferase 2B (also known as HRX2; MLL2; MLL4; TRX2; WBP7; DYT28; MLL1B; MRD68; WBP-7; and CXXC10), and the protein contains a CXXC zinc finger, three PHD zinc fingers, two FY-rich domains, and a SET domain. In some embodiments, KMT2B can be a human (gene ID: 9757), non-human primate (e.g., gene ID: 102115861), or rodent gene (e.g., gene ID: 75410, gene ID: 102550344). In humans, mutations in the gene encoding KMT2B are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated by, for example, GenBank RefSeq accession numbers: NM_014727.3; XM_011527561.3; XM_011527562.3; XM_047439787.1; and XR_935878.3) have been characterized.

[0151] Krabbe disease: As used herein, the terms "Krabbe disease", "KD", "KRD", "globoid cell leukodystrophy", or "galactosylceramide lipidosis" refer to a metabolic disorder in which lipids accumulate to harmful levels in different tissues throughout the body, including the nervous system, resulting in the death of cells in the central nervous system, including the brain. Krabbe disease is characterized by cells having more than one nucleus (globoid cells), which results in the breakdown of the myelin sheath on nerves. Krabbe disease results from a mutation in the GALC gene that causes a deficiency of the galactosylceramidase enzyme. Galactosylceramidase is an essential enzyme in myelin metabolism. The symptoms of Krabbe disease include irritability, a rigid posture, mental retardation, physical developmental delay, deterioration of motor skills, muscle weakness, muscle hypertonia, myoclonic seizures, spasticity, fever, blindness, difficulty swallowing, and hearing loss. Krabbe disease is most commonly found in infants (infantile form) and usually begins before the age of 1 year. 10% - 15% of Krabbe disease patients have a late onset of this disease, which occurs in the juvenile or adult form. In some embodiments, the subject has a mutation in the GALC gene. In some embodiments, the subject has an accumulation of lipids in the central nervous system (e.g., the brain). In some embodiments, the subject has globoid cells in the central nervous system (e.g., the brain). In some embodiments, the subject has a deficiency of the galactosylceramidase enzyme. In some embodiments, the subject has one or more of the following symptoms: irritability, a rigid posture, mental retardation, physical developmental delay, deterioration of motor skills, muscle weakness, muscle hypertonia, myoclonic seizures, spasticity, fever, blindness, difficulty swallowing, and hearing loss.

[0152] LRRK2: As used herein, LRRK2 refers to the gene encoding the dardarin protein, also known as leucine-rich repeat kinase 2 and PARK8. Variants of LRRK2 are associated with an increased risk of Parkinson's disease. Mutations in LRRK2 encoding the G2019S mutant of the dardarin protein have been shown to cause Parkinson's disease and are a relatively common cause of familial Parkinson's disease. This G2019S mutation results in enhanced kinase activity of the protein. Mutations in LRRK2 are the most common known cause of both familial and sporadic Parkinson's disease.

[0153] LSD: As used herein, the term "LSD" refers to the gene encoding the lysine-specific demethylase of proteins involved in neuronal differentiation and physiology. One example of LSD is LSD1 (also known as KDM1A, AOF2, BHC110, CPRF, and KDM1). In some embodiments, LSD can be a human (Gene ID: 23028), non-human primate (e.g., Gene ID: 718609), or rodent gene (e.g., Gene ID: 99982, Gene ID: 500569). In humans, mutations in the gene encoding LSD are associated with neurodegeneration such as Alzheimer's disease, tauopathy, and / or frontotemporal dementia. In addition, multiple human transcript variants of LSD such as LSD1 encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_015013.4 and NM_001009999.3) have been characterized.

[0154] MAPT: When used herein, the term "MAPT" refers to the gene encoding the microtubule-associated protein tau (also known as tau, tau-40, DDPAC, FTDP-17, MAPTL, MSTd, MTBT1, MTBT2, PPND, and PPP1R103), which is involved in the stabilization of axonal microtubules. In some embodiments, MAPT can be a human (Gene ID: 4137), non-human primate (e.g., Gene ID: 574327), or rodent gene (e.g., Gene ID: 17762, Gene ID: 29477). In humans, mutations in the MAPT gene can be associated with the development of Alzheimer's disease. Mutations in the MAPT gene can also be associated with certain tauopathies, including frontotemporal dementia. Aggregates formed by hyperphosphorylated tau protein contribute to the pathology of Alzheimer's disease. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_016835.5, NM_005910.6, and NM_001377265.1) have been characterized.

[0155] MECP2: As used herein, the term MECP2 refers to the gene encoding methyl-CpG binding protein 2, a protein that binds to methylated DNA and has an important role in mammalian development (also known as RS; RTS; RTT; PPMX; MRX16; MRX79; MRXSL; AUTSX3; MRXS13). In some embodiments, MECP2 can be a human (Gene ID: 4204), non-human primate (e.g., Gene ID: 102135563), or rodent gene (e.g., Gene ID: 17257, Gene ID: 29386). In humans, mutations in the MECP2 gene are the cause of most cases of Rett syndrome. Multiple human transcript variants encoding different protein isoforms (as annotated, e.g., by GenBank RefSeq accession numbers: NM_004992.4; NM_001110792.2; NM_001316337.2; NM_001369391.2; NM_001369392.2; NM_001369393.2; NM_001369394.2; NM_001386137.1; NM_001386138.1; NM_001386139.1) have been characterized. Suppression of MECP2 (e.g., in its mutant form) may be effective in treating Rett syndrome. Increasing the level and / or activity of methyl-CpG binding protein 2 or a functional fragment thereof may also be effective in treating Rett syndrome.

[0156] Metachromatic leukodystrophy (MLD): As used herein, the term "metachromatic leukodystrophy" or "MLD" refers to a lysosomal storage disease (LSD) characterized by a deficiency in the lysosomal enzyme arylsulfatase A (ARSA) or its sphingolipid activator protein B (SapB), which causes dysfunction and destruction of myelin sheaths in the central and peripheral nervous systems. This leads to a progressive deterioration of neurodevelopment and neurocognitive function. Metachromatic leukodystrophy is associated with mutations in the arylsulfatase A gene (ARSA) and / or the prosaposin gene (PSAP). Currently, there is no treatment effective against metachromatic leukodystrophy. In some embodiments, a subject (e.g., a subject diagnosed with MLD) has deterioration of myelin sheaths. In some embodiments, a subject (e.g., a subject diagnosed with MLD) has deterioration of neurodevelopment and / or neurocognitive function.

[0157] Molecular payload: As used herein, the term "molecular payload" refers to a molecule or species that functions to modulate biological fate. In some embodiments, the molecular payload is linked or otherwise associated with a CNS targeting agent. In some embodiments, the molecular payload is a small molecule, polypeptide (e.g., protein, peptide, antibody), gene therapy payload (e.g., nucleic acid), or oligonucleotide. In some embodiments, the molecular payload functions to regulate transcription of a DNA sequence, to regulate expression of a protein, or to regulate the activity of a protein. In some embodiments, the molecular payload is an oligonucleotide comprising a strand having a region of complementarity to a target gene. In some embodiments, the molecular payload is a polypeptide having biological activity in the context of a particular disease (e.g., a CNS disease or disorder). In some embodiments, the molecular payload is a small molecule having biological activity in the context of a particular disease (e.g., a CNS disease or disorder). In some embodiments, the molecular payload is a gene therapy payload encoding a bioactive compound (e.g., a polypeptide).

[0158] Motor neuron disease: As used herein, the term “motor neuron disease” refers to a group of progressive neurological disorders that destroy motor neurons that control skeletal muscle activity. Motor neuron diseases include diseases such as ALS, progressive bulbar palsy, primary lateral sclerosis, progressive muscular atrophy, spinal muscular atrophy, Kennedy disease, and post-polio syndrome. Various genes and mutations in them are associated with the occurrence of motor neuron diseases. For example, APOE (e.g., the APOE4 allele) is associated with the occurrence of certain types of motor neuron diseases. Thus, in some aspects, allele-specific regulation of APOE (e.g., APOE4) is useful for the treatment of motor neuron diseases. In some aspects, a subject in need of treatment for a motor neuron disease has one or more symptoms associated therewith.

[0159] MSH3: As used herein, MSH3 refers to the gene encoding the MutS homolog 3 protein. This protein is involved in the mismatch repair system. MSH3 has a significant role in tumor suppression by repairing somatic mutations on DNA in cancer, and both loss and overexpression of MSH3 expression can lead to oncogenic effects. Overexpression of MSH3 has been shown to decrease mismatch repair ability, and increased expression of MSH3 is associated with the progression of Huntington's disease. See, for example, Flower, et al. “MSH3 modifies somatic instability and disease severity in Huntington’s and myotonic dystrophy type 1” Brain 142(7):1876-1888 (2019). The evidence also suggests that mutations in MSH3 may be associated with the occurrence of spinocerebellar ataxia.

[0160] Niemann-Pick type A: As used herein, the term "Niemann-Pick type A" refers to Niemann-Pick disease type A (NPA), also known as infantile visceral neurovisceral acid sphingomyelinase deficiency. This is a fatal lysosomal neurodegenerative disorder associated with a deficiency in the activity of acid sphingomyelinase. NPA is inherited in an autosomal recessive pattern. The symptoms of NPA include developmental delay, hepatosplenomegaly, lung injury, visual abnormalities, neurodegeneration, and early death. The NPA disease is characterized by the accumulation of sphingomyelin in lysosomes, dysfunction of the autophagy-lysosomal pathway, and astrogliosis. The genetic cause of the NPA disease has been identified as a mutation in the ASM gene. See, for example, Marin, et al. “c-Abl activation linked to autophagy-lysosomal dysfunction contributes to neurological impairment in Niemann-Pick type A disease” Front Cell Devel Biol. 10: 844297 (2022).

[0161] Oligonucleotide: As used herein, the term "oligonucleotide" refers to an oligomeric nucleic acid compound up to 200 nucleotides in length. Examples of oligonucleotides include, but are not limited to, RNAi oligonucleotides (e.g., siRNA, shRNA), microRNAs, gapmers, mixmers, phosphorodiamidate morpholinos, peptide nucleic acids, aptamers, guide nucleic acids (e.g., Cas9 guide RNA), etc. The oligonucleotide can be single-stranded or double-stranded. In some embodiments, the oligonucleotide can include one or more modified nucleosides (e.g., 2'-O-methyl sugar modification, purine or pyrimidine modification). In some embodiments, the oligonucleotide can include one or more modified internucleoside linkages. In some embodiments, the oligonucleotide can include one or more phosphorothioate linkages, which can be in the Rp or Sp stereochemical conformation.

[0162] Parkinson's disease: Parkinson's disease is a neurological disorder that primarily affects the motor system and is a form of synucleinopathy associated with abnormal accumulation of the protein alpha-synuclein in the brain. The motor symptoms of the disease result from the death of cells in the substantia nigra region of the midbrain, which leads to a deficiency of dopamine. The cause of cell death is poorly understood, but it involves the accumulation of misfolded proteins in Lewy bodies in neurons. At least 11 autosomal dominant and 9 autosomal recessive gene mutations are involved in the development of Parkinson's disease, including mutations in SNCA, LRRK2, PARK3, UCHL1, GIGYF2, HTRA2, EIF4G1, TMEM230, CHCHD2, RIC3, VPS35 (autosomal dominant); and PRKN, PINK2, PARK7, ATP13A2, PLA2G6, FBXO7, DNAJC6, SYNJ1, and VPS13C (autosomal recessive). Mutations in SNCA and LRRK2 have been found to be risk factors for sporadic Parkinson's disease.

[0163] PCDH19: As used herein, the term "PCDH19" refers to the gene encoding the protein protocadherin 19 (also referred to as DEE9, EFMR, and EIEE9) that is involved in cell adhesion. In some embodiments, PCDH19 can be a human (Gene ID: 57526), non-human primate (e.g., Gene ID: 703042), or rodent gene (e.g., Gene ID: 279653, Gene ID: 317183). In humans, mutations in the gene encoding PCDH19 are associated with epilepsy and seizures. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_020766.3 and NM_001184880.2) have been characterized.

[0164] PIKFYVE: As used herein, PIKFYVE refers to the gene encoding phosphatidylinositol 3-phosphate 5-kinase type III protein (PIPKIII) (also known as CFD; FAB1; HEL37; PIP5K; PIP5K3; ZFYVE29), which phosphorylates certain phosphatidylinositols (e.g., PtdIns and PtdIns3P). In some embodiments, PIKFYVE can be a human (gene ID: 200576), non-human primate (e.g., gene ID: 710115), or rodent gene (e.g., gene ID: 18711, gene ID: 316457). PIKFYVE and mutations therein are involved in ALS and frontotemporal dementia (FTD). Multiple human transcript variants encoding different protein isoforms (as annotated, e.g., by GenBank RefSeq accession numbers: NM_015040.4; NM_152671.4; NM_001178000.2) have been characterized.

[0165] PNKD: As used herein, PNKD refers to a protein thought to play a role in the control of myofibrillogenesis, the gene encoding PNKD metallo-beta-lactamase domain-containing (also referred to as R1; MR1; PDC; DYT8; FPD1; MR-1; BRP17; MR-1S; PKND1; PNKD1; FKSG19; TAHCCP2; KIPP1184). In some embodiments, PNKD can be a human (gene ID: 25953), non-human primate (e.g., gene ID: 101867223), or rodent gene (e.g., gene ID: 56695, gene ID: 100188944). In humans, mutations in the gene encoding PNKD are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_015488.5; NM_022572.4; NM_001077399.3; XM_017003771.2; and XM_017003772.2) have been characterized.

[0166] PRKRA: As used herein, PRKRA refers to the gene encoding the protein activator (also known as RAX; PACT; DYT16; HSD14) of protein kinase EIF2AK2, an interferon-inducible protein kinase that is activated by double-stranded RNA that mediates the effects of interferon in response to viral infection. In some embodiments, PRKRA can be a human (gene ID: 8575), non-human primate (e.g., gene ID: 102116511), or rodent gene (e.g., gene ID: 23992, gene ID: 311130). In humans, mutations in the gene encoding PRKRA are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_003690.5; NM_001139517.1; NM_001139518.1; NM_001316362.2; XM_011512063.3; and XM_047446138.1) have been characterized.

[0167] PrP: As used herein, the term "PrP" refers to a protein involved in neural function that can form cytotoxic prions, the gene encoding the prion protein (also referred to as PRNP, PRIP, CD230, and CJD). In some embodiments, PrP can be a human (Gene ID: 5621), non-human primate (e.g., Gene ID: 717859), or rodent gene (e.g., Gene ID: 19122, Gene ID: 24686). In humans, mutations in the PrP gene are associated with neurodegeneration. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000311.5 and NM_183079.4) have been characterized. In some embodiments, PrP is associated with small fiber neuropathy, phenotypes related to nociception, Alexander disease, Angelman syndrome, autism spectrum disorder, retinitis pigmentosa, isolated macular dystrophy, and / or multiple sclerosis.

[0168] PRRT2: As used herein, PRRT2 refers to the gene encoding a transmembrane protein that contains a proline-rich domain in its N-terminal half, proline-rich transmembrane protein 2 (also known as PKC; EKD1; ICCA; BFIC2; BFIS2; DSPB3; DYT10; FICCA; IFITMD1). In some embodiments, PRRT2 can be a human (gene ID: 112476), non-human primate (e.g., gene ID: 102124815), or rodent gene (e.g., gene ID: 69017, gene ID: 361651). In humans, mutations in the gene encoding PRRT2 are associated with the development of hereditary dystonia. Additionally, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_145239.3; NM_001256442.2; NM_001256443.2; XM_011545715.4; XM_017022887.3; XM_017022888.3; and XM_017022889.3) have been characterized.

[0169] Recombinant antibody: As used herein, the term "recombinant human antibody" is intended to include all human antibodies prepared, expressed, made, or isolated by recombinant means, such as antibodies expressed using recombinant expression vectors transfected into host cells (described in more detail in this disclosure), antibodies isolated from recombinant combinatorial human antibody libraries (Hoogenboom H. R., (1997) TIB Tech. 15:62-70; Azzazy H., and Highsmith W. E., (2002) Clin. Biochem. 35:425-445; Gavilondo J. V., and Larrick J. W. (2002) BioTechniques 29:128-145; Hoogenboom H., and Chames P. (2000) Immunology Today 21:371-378), antibodies isolated from animals transgenic for human immunoglobulin genes (e.g., mice) (see, for example, Taylor, L. D., et al. (1992) Nucl. Acids Res. 20:6287-6295; Kellermann S-A., and Green L. L. (2002) Current Opinion in Biotechnology 13:593-597; Little M. et al (2000) Immunology Today 21:364-370), or antibodies prepared, expressed, made, or isolated by any other means involving splicing of human immunoglobulin gene sequences to other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when transgenic animals for human Ig sequences are used, in vivo somatic mutagenesis), and thus, the amino acid sequences of the VH and VL regions of the recombinant antibody are derived from and related to human germline VH and VL sequences but are sequences that may not naturally occur within the in vivo human antibody germline repertoire.One aspect of the present disclosure provides a fully human antibody that can bind to the human transferrin receptor, which can be made using techniques well known in the art. For example, a human Ig phage library such as that disclosed in Jermutus et al., PCT Publication No. WO 2005 / 007699 A2 is used, but is not limited thereto.

[0170] Complementary region: As used herein, the term "complementary region" refers to the nucleotide sequence of an oligonucleotide that is sufficiently complementary, as an example, to the corresponding nucleotide sequence of a target nucleic acid, such that the two nucleotide sequences can anneal to each other under physiological conditions (e.g., intracellularly). In some aspects, the complementary region is completely complementary to the corresponding nucleotide sequence of the target nucleic acid. However, in some aspects, the complementary region is partially complementary to the corresponding nucleotide sequence of the target nucleic acid (e.g., at least 80%, 90%, 95%, or 99% complementary). In some aspects, the complementary region contains 1, 2, 3, or 4 mismatches compared to the corresponding nucleotide sequence of the target nucleic acid.

[0171] Rett syndrome: As used herein, the term "Rett syndrome" refers to the spectrum of disorders associated with mutations in MECP2. Rett syndrome is a brain disorder that occurs almost exclusively in female children. Around 6 to 18 months of age, subjects with Rett syndrome begin to develop severe problems with language and communication, learning, coordination, and other brain functions. Early in childhood, affected subjects lose their intentional use of their hands and begin to perform repetitive hand wringing, hand washing, or clapping motions. They tend to grow more slowly than other children, and approximately 75% have microcephaly. Other possible signs and symptoms include abnormal breathing, coughing or drooling, abnormal eye movements such as intense staring or excessive blinking, cold hands and feet, irritability, sleep disorders, seizures, and scoliosis. More than 99% of subjects with Rett syndrome do not have a family history of the disease; many of these cases are due to new mutations in MECP2. As an example, suppression of mutant forms of MECP2 by antisense oligonucleotide therapy may be effective in treating Rett syndrome or its symptoms in some aspects. Increasing the level and / or activity of the protein encoded by MECP2 or a functional fragment thereof (e.g., by delivery of a gene therapy payload) may also be effective in treating Rett syndrome or its symptoms.

[0172] Sandhoff disease: As used herein, the term "Sandhoff disease," which is sometimes referred to as GM2 gangliosidosis, refers to a series of disorders that progressively damage neurons in the central nervous system. Sandhoff disease is inherited in an autosomal recessive pattern. The symptoms of Sandhoff disease include progressive weakness, seizures, lack of development, neurological disorders, cognitive impairment, and early death. Sandhoff disease is characterized by cortical, cerebellar, and spinal cord atrophy. The genetic cause of Sandhoff disease has been identified as approximately 30 mutations in the HEXB gene. See Xiao, et al. “Sandhoff Disease” 2022 Apr 14, in: Adam MP, Mirzaa GM, Pagon RA, et al., editors. GeneReviews (R) [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2023. In some embodiments, a subject in need of treatment for Sandhoff disease exhibits progressive weakness. In some embodiments, the progressive weakness includes lower limb weakness. In some embodiments, a subject in need of treatment for Sandhoff disease exhibits seizures. In some embodiments, a subject in need of treatment for Sandhoff disease exhibits lack of development. In some embodiments, the lack of development includes developmental plateauing and developmental regression. In some embodiments, a subject in need of treatment for Sandhoff disease exhibits neurological disorders. In some embodiments, the neurological disorders include loss of motor skills, excessive startle response, hypotonia, hyperreflexia, neuropathy, neuronopathy, atrophy, fasciculation, balance problems, tremors, dysarthria, dysphagia, and spasticity. In some embodiments, a subject in need of treatment for Sandhoff disease exhibits cognitive impairment. In some embodiments, the cognitive impairment includes decreased attention, reduced cognitive function, lack of executive function, and lack of memory. In some embodiments, a subject in need of treatment for Sandhoff disease does not exhibit hepatosplenomegaly. Sandhoff disease is very similar to Tay-Sachs; thus, treatments for Tay-Sachs may also be effective in treating Sandhoff disease (and vice versa).Sandhoff disease is also closely related to GM1 gangliosidosis, and thus, treatments for GM1 gangliosidosis may be effective in treating Sandhoff disease.

[0173] SCA1: As used herein, "SCA1" refers to spinocerebellar ataxia type 1 associated with a CAG repeat expansion in ATXN1. See "spinocerebellar ataxia."

[0174] SCA2: As used herein, "SCA2" refers to spinocerebellar ataxia type 2 associated with a CAG repeat expansion in ATXN2. See "spinocerebellar ataxia."

[0175] SCA3: As used herein, "SCA3" refers to spinocerebellar ataxia type 3 associated with a CAG repeat expansion in ATXN3. See "spinocerebellar ataxia."

[0176] SCN1A: As used herein, the term "SCN1A" refers to the gene (also referred to as DEE6, DEE6A, DEE6B, DRVT, EIEE6, FEB3, FEB3A, FHM3, GEFSP2, HBSCI, NAC1, Nav1.1, SCN1, and SMEI) that encodes a protein involved in the generation and propagation of action potentials in neurons. In some embodiments, SCN1A can be a human (Gene ID: 6323), non-human primate (e.g., Gene ID: 704086), or rodent gene (e.g., Gene ID: 20265, Gene ID: 81574). In humans, mutations in SCN1A, such as loss-of-function mutations in SCN1A, are associated with epilepsy and seizures, as well as Dravet syndrome (severe myoclonic epilepsy in infancy (SMEI)). Gain-of-function mutations in SCN1A are associated with other neurological disorders such as familial hemiplegic migraine, epileptic encephalopathy, and arthrogryposis. See, for example, Brunklaus, et al. “The gain of function SCN1A disorder spectrum: novel epilepsy phenotypes and therapeutic implications” Brain 145(11): 3816-3831 (2022) and Ding, et al. “SCN1A Mutation-Beyond Dravet Syndrome: A Systematic Review and Narrative Synthesis” Front Neurol. 12: 743726 (2021). The most common mutations in the SCN1A gene include Thr226Met, Leu263Val, Val422Leu, Thr1174Ser, Trp1204Arg, Pro1345Ser, Gln1489Lys, Phe1499Leu, Arg1575Cys, Val1611Phe, Leu1624Pro, Arg1648Cys, Leu1649Gln, Leu1670Trp, Gly1674Arg, and Asp1866Tyr.Mutations in SCN1A often result in reduced function of the encoded protein or no protein expression. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_006920.6 and NM_001165963.4) have been characterized. In some embodiments, SCN1A or its mutant forms are associated with painful disorders.

[0177] SCN2A: When used herein, the term "SCN2A" refers to the gene encoding the protein involved in the generation and propagation of action potentials in neurons, voltage-gated sodium channel alpha subunit 2 (also referred to as BFIC3, BFIS3, BFNIS, DEE11, EA9, EIEE11, HBA, HBSCI, HBSCII, NAC2, Nav1.2, SCN2A1, and SCN2A2). In some embodiments, SCN2A can be a human (Gene ID: 6326), non-human primate (e.g., Gene ID: 703298), or rodent gene (e.g., Gene ID: 110876, Gene ID: 24766). In humans, mutations in the SCN2A gene, such as gain-of-function mutations in SCN2A, are associated with epilepsy and seizures. Loss-of-function mutations in SCN2A are associated with other neurological disorders, including autism spectrum disorder with or without epilepsy. See, for example, Zeng, et al. “SCN2A-Related Epilepsy: The Phenotypic Spectrum, Treatment and Prognosis” Front Mol Neurosci. 15: 809951 (2022) doi: 10.3389 / fnmol.2022.809951. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_021007.3 and NM_001040142.2) have been characterized. In some embodiments, SCN2A or its mutant forms are associated with pain disorders.

[0178] SCN8A: As used herein, the term "SCN8A" refers to the gene (also referred to as BFIS5, CERIII, CIAT, DEE13, EIEE13, MED, MYOCL2, NaCh6, Nav1.6, and PN4) that encodes the protein involved in the generation and propagation of action potentials in neurons. In some embodiments, SCN8A can be a human (Gene ID: 6334), non-human primate (e.g., Gene ID: 695972), or rodent gene (e.g., Gene ID: 20273, Gene ID: 29710). In humans, mutations in the SCN8A gene, such as gain-of-function mutations in SCN8A, are associated with epilepsy and seizures. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_014191.4 and NM_001330260.2) have been characterized. In some embodiments, SCN8A or its mutant forms are associated with painful disorders.

[0179] SCN9A: As used herein, the term "SCN9A" refers to the gene (also referred to as ETHA, FEB3B, GEFSP7, HSAN2D, NE-NA, NENA, Nav1.7, PN1, and SFNP) that encodes a protein involved in the generation and propagation of action potentials in neurons, the voltage-gated sodium channel alpha subunit 9. In some embodiments, SCN9A can be a human (Gene ID: 6335), non-human primate (e.g., Gene ID: 574119), or rodent gene (e.g., Gene ID: 20274, Gene ID: 78956). In humans, mutations in the SCN9A gene are associated with various painful disorders. In some embodiments, mutations in SCN9A are associated with phenotypes related to small fiber neuropathy and nociception. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_002977.3 and NM_001365536.1) have been characterized.

[0180] SGCE: As used herein, SGCE refers to the gene encoding sarcoglycan epsilon, a member of the sarcoglycan family (also referred to as ESG; DYT11; epsilon-SG). Sarcoglycans are transmembrane proteins that are components of the dystrophin-glycoprotein complex, which links the actin cytoskeleton to the extracellular matrix. Unlike other family members that are predominantly expressed in striated muscle, epsilon sarcoglycan is more widely expressed. In some embodiments, SGCE can be a human (Gene ID: 8910), non-human primate (e.g., Gene ID: 101865326), or rodent gene (e.g., Gene ID: 20392, Gene ID: 432360). In humans, mutations in the gene encoding SGCE are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession numbers: NM_003919.3; NM_001099400.2; NM_001099401.2; NM_001301139.2; NM_001346713.2; NM_001346715.2; NM_001346717.2; NM_001346719.2; NM_001346720.2; NM_001362807.2; NM_001362808.2; and NM_001362809.2) have been characterized.

[0181] SLC2A1: As used herein, SLC2A1 refers to the gene encoding the major glucose transporter at the mammalian blood-brain barrier, solute carrier family 2 member 1 (also referred to as CSE; PED; DYT9; GLUT; DYT17; DYT18; EIG12; GLUT1; HTLVR; GLUT-1; SDCHCN; GLUT1DS). In some embodiments, SLC2A1 can be a human (Gene ID: 6513), non-human primate (e.g., Gene ID: 102144217), or rodent gene (e.g., Gene ID: 20525, Gene ID: 24778). In humans, mutations in the gene encoding SLC2A1 are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, e.g., as GenBank RefSeq accession number: NM_006516.4) have been characterized.

[0182] SMN: As used herein, SMN refers to the gene encoding the survival of motor neuron protein. The survival of motor neuron protein is involved in transcriptional splicing through its involvement in the assembly of ribonucleoprotein that binds to pre-mRNA to form spliceosomes. Lack of survival of motor neuron protein activity results in widespread splicing defects, particularly in spinal motor neurons, and degeneration of lower motor neurons in the spinal cord. The survival of motor neuron protein is encoded by the genes SMN1 and SMN2, mutations in each of which are associated with spinal muscular atrophy, and both of which can be referred to as "SMN". Molecular payloads useful for modulating SMN1 may also be useful for modulating SMN2 in the treatment of CNS diseases and disorders, and vice versa.

[0183] SNCA: As used herein, SNCA refers to the gene encoding the alpha-synuclein protein. Alpha-synuclein is a neuronal protein that regulates synaptic vesicle trafficking and subsequent neurotransmitter release and is abundant in the brain. SNCA is expressed primarily in neural tissue (e.g., neurons), but can also be found in glial cells. Alpha-synuclein is found predominantly in both free and membrane-bound forms at presynaptic terminals, and approximately 15% of the protein is membrane-bound at any given time in neurons. Alternative splicing of the SNCA transcript results in the production of at least three isoforms of alpha-synuclein. In pathological conditions characterized by the presence of Lewy bodies, including Parkinson's disease, alpha-synuclein aggregates to form insoluble fibrils. These pathological conditions are known as synucleinopathies. The mechanism of aggregation of alpha-synuclein is unknown. Some mutations in SNCA are associated with Parkinson's disease and include mutations that result in alpha-synuclein proteins with amino acid substitutions A53T, A53T, A30P, E46K, H50Q, G51D, A18T, A29S, A53E, A53V, E57A, V15A, T72M, L8I, V15D, M127I, P117S, M5T, G93A, E83Q, and A30G. The alpha-synuclein protein has been shown to interact with the dopamine transporter, parkin (ligase), phospholipase D1, SNCAIP, tau protein, and beta-amyloid.

[0184] SOD1: The term "SOD1" refers to the enzyme superoxide dismutase 1 and the gene encoding it. SOD1 is an enzyme involved in apoptosis, amyotrophic lateral sclerosis (ALS), and Parkinson's disease. The SOD1 protein is a 32 kDa homodimer containing binuclear Cu / Zn sites on each subunit. The Cu / Zn sites are responsible for breaking down free superoxide radicals in the body by catalyzing the disproportionation of superoxide to hydrogen peroxide and dioxygen. Wild-type SOD1 protein demonstrates anti-apoptotic properties in nerve culture, while mutant SOD1 protein has been shown to promote apoptosis in neurons. Mutations in the SOD1 gene are linked to familial ALS, but wild-type SOD1 is also involved in a significant proportion of sporadic ALS cases, which account for 90% of ALS patients. The most frequent SOD1 mutations are A4V, H46R, and G93S. Virtually all known ALS-related SOD1 mutations act in a dominant manner, such that a single mutant copy of the SOD1 gene is sufficient to cause the disease. The exact mechanism by which mutations in SOD1 cause ALS is unknown, but some evidence suggests that it is the result of a toxic gain of function. This is because many disease-related SOD1 mutations (including A4V and G93A) retain enzyme activity, and Sod1-deficient mice do not develop ALS. The DNA oxidation product 8-OHdG, a well-established marker of oxidative DNA damage, accumulates in the mitochondria of motor neurons in ALS patients, suggesting that oxidative damage to the DNA (such as mitochondrial DNA) of motor neurons resulting from mutant SOD1 may significantly contribute to the etiology of ALS.

[0185] Specific binding: As used herein, the term "specific binding" refers to the ability of a molecule to bind to a binding partner with a degree of affinity or avidity that allows the molecule to be used to distinguish the binding partner from an appropriate control in a binding assay or other binding context. With respect to an antibody, the term "specific binding" refers to the ability of an antibody to bind to a specific antigen with a degree of affinity or avidity that allows the antibody to be used to distinguish the specific antigen from others, as compared to an appropriate reference antigen(s), to the extent that, for example, preferential targeting to certain cells, for example CNS cells, through binding to the antigen, as described herein. In some embodiments, an antibody has a binding affinity of at least about 10 -4 M, 10 -5 M, 10 -6 M, 10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, 10 -12 M, 10 -13 M, or K for binding to fewer targets D In some embodiments, the antibody specifically binds to a target when it has the following structure: In some embodiments, the antibody specifically binds to a transferrin receptor, e.g., an epitope of the apical domain of the transferrin receptor.

[0186] Spinocerebellar ataxia: As used herein, the term "spinocerebellar ataxia" or "SCA" refers to a class of CNS disorders generally characterized by coordination problems caused by effects on the cerebellum and spinal cord (also sometimes referred to as "autosomal dominant cerebellar ataxia"). SCA often features a slowly progressive coordination disorder of walking and is often associated with poor coordination of the hands, speech, and eye movements. SCA is a progressive neurodegenerative disorder that follows an autosomal dominant pattern of inheritance. There are more than 40 types of SCA, each of which has similar causes and symptoms. The most common form of SCA is SCA3, also known as Machado-Joseph disease. Most genetic mutations associated with SCA result in significant damage to cerebellar Purkinje neurons with continuous cerebellar atrophy. In addition, other parts of the CNS, such as the spinal cord, basal ganglia, and pontine nuclei in the brainstem, may be involved. As an example, see Klockgether, et al. “Spinocerebellar ataxia” Nat Rev Dis Primers 5:24 (2019) doi:10.1038 / s41572-019-0074-3 and Bhandari, et al. “Spinocerebellar Ataxia.” [Updated 2022 Aug 10]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; Available from: ncbi.nlm.nih.gov / books / NBK557816 / . SCA1, SCA2, SCA3, and SCA related to MSH3 are particularly relevant to the present disclosure. Many forms of SCA are associated with an expansion of trinucleotide repeats within the ataxin (ATXN) genes. SCA1 is associated with a CAG repeat expansion in ATXN1, SCA2 is associated with a CAG repeat expansion in ATXN2, and SCA3 is associated with a CAG repeat expansion in ATXN3. Currently, there is no treatment available for SCA; thus, clinical intervention focuses primarily on symptom management through physical therapy, occupational therapy, and speech and language therapy.

[0187] SPR: As used herein, SPR refers to the gene encoding aldoketoreductase, sepiapterin reductase, which catalyzes the NADPH-dependent reduction of pteridine derivatives (also referred to as SDR38C1). SPR is important for the biosynthesis of tetrahydrobiopterin (BH4). Mutations in this gene result in DOPA-responsive dystonia due to sepiapterin reductase deficiency. In some embodiments, SPR can be a human (Gene ID: 6697), non-human primate (e.g., Gene ID: 102128831), or rodent gene (e.g., Gene ID: 20751, Gene ID: 29270). In humans, mutations in the gene encoding SPR are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, as in GenBank RefSeq accession number: NM_003124.5) have been characterized.

[0188] Subject: As used herein, the term "subject" refers to a mammal. In some embodiments, the subject is a non-human primate or a rodent. In some embodiments, the subject is a human. In some embodiments, the subject is a patient, e.g., a human patient having or suspected of having a disease. In some embodiments, the subject is a human patient having or suspected of having a CNS disease or disorder. In some embodiments, the subject is a human patient having one or more symptoms associated with a CNS disease or disorder, e.g., one or more symptoms disclosed herein.

[0189] SYF2: As used herein, SYF2 refers to the gene encoding the pre-mRNA splicing factor SYF2 that is primarily localized in the nucleus (also known as P29, CBPIN, NTC31, fSAP29). In some embodiments, SYF2 can be a human (Gene ID: 25949), non-human primate (e.g., Gene ID: 102139055), or rodent gene (e.g., Gene ID: 68592, Gene ID: 170933). SYF2 and mutations therein are involved in ALS and frontotemporal dementia (FTD). Multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_207170.4; NM_015484.5) have been characterized.

[0190] TAF1: As used herein, TAF1 refers to the gene encoding TATA box binding protein associated factor 1 (also referred to as OF; XDP; BA2R; CCG1; CCGS; DYT3; KAT4; P250; NSCL2; TAF2A; MRXS33; N-TAF1; TAFII250; DYT3 / TAF1; TAFII-250; TAF(II)250), which is a member of a group of evolutionarily conserved proteins known as TBP-associated factors. TAF1 encodes the largest subunit of the basal transcription factor TFIID, and this subunit binds to the core promoter sequence encompassing the transcription start site and also binds to activators and other transcriptional regulatory factors. In some embodiments, TAF1 can be a human (Gene ID: 6872), non-human primate (e.g., Gene ID: 102118965), or rodent gene (e.g., Gene ID: 270627, Gene ID: 317256). In humans, mutations in the gene encoding TAF1 are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_004606.5; NM_138923.4; NM_001286074.2; NR_104387.2; NR_104388.2; NR_104389.2; NR_104390.2; NR_104391.2; NR_104392.2; NR_104393.2; NR_104394.2; NR_104395.2; NR_104396.2; XM_005262300.3; XM_024452430.2; XM_047442391.1; XM_047442392.1; XM_047442393.1; XM_047442394.1; XM_047442395.1; XM_047442396.1; XM_047442397.1; XM_047442398.1; XM_047442399.1; XM_047442400.1; XM_047442401.1; XM_047442402.1; XM_047442403.1; XM_047442404.1; XM_047442405.1; and XM_047442406.1) have been characterized.

[0191] Tay-Sachs: As used herein, the term "Tay-Sachs" refers to a genetic disorder characterized by the destruction of nerve cells in the central nervous system. Tay-Sachs is also known as GM2 gangliosidosis. Tay-Sachs is associated with mutations in the enzyme hexosaminidase A (HEXA), which leads to the accumulation of GM2 ganglioside in lysosomes and nerve cells. Tay-Sachs overwhelmingly affects infants (infantile form), but can also occur in adolescence (juvenile form) and adulthood. Tay-Sachs is characterized by neurodegeneration, and its symptoms include: slow development, progressive loss of intellectual ability, dementia, blindness, increased startle reflex to noise, progressive loss of hearing, swallowing problems, seizures, cherry-red spot in the eye, muscle weakness, and ataxia. Conventional treatment of Tay-Sachs focuses on symptom relief and delaying progression. In some embodiments, a subject in need of treatment for Tay-Sachs has a mutation in the HEXA gene. In some embodiments, a subject in need of treatment for Tay-Sachs has neurodegeneration. In some embodiments, a subject in need of treatment for Tay-Sachs has one or more of the following symptoms: slow development, progressive loss of intellectual ability, dementia, blindness, increased startle reflex to noise, progressive loss of hearing, swallowing problems, seizures, cherry-red spot in the eye, muscle weakness, and ataxia. Tay-Sachs is very similar to Sandhoff disease; thus, treatment for Sandhoff disease may also be effective in treating Tay-Sachs (and vice versa). Tay-Sachs is also closely related to GM1 gangliosidosis; thus, treatment for GM1 gangliosidosis may be effective in treating Tay-Sachs (and vice versa).

[0192] TH: As used herein, TH refers to the gene (also referred to as TYH; DYT14; DYT5b) that encodes the protein tyrosine hydroxylase, which is involved in the conversion of tyrosine to dopamine. The tyrosine hydroxylase protein is the rate-limiting enzyme in the synthesis of catecholamines and thus plays a key role in the physiology of adrenergic neurons. In some embodiments, TH can be a human (gene ID: 7054), non-human primate (e.g., gene ID: 102134074), or rodent gene (e.g., gene ID: 21823, gene ID: 25085). In humans, mutations in the gene encoding TH are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated by, for example, GenBank RefSeq accession numbers: NM_000360.4; NM_199292.3; NM_199293.3; and XM_011520335.3) have been characterized.

[0193] THAP1: As used herein, THAP1 refers to the gene (also referred to as DYT6) that encodes the protein containing the THAP domain of the conserved DNA-binding domain, THAP domain-containing 1. This protein co-localizes with the apoptosis response protein PAWR / PAR-4 in the promyelocytic leukemia (PML) nuclear bodies and functions as an apoptosis-promoting factor that links PAWR to the PML nuclear bodies. In some embodiments, THAP1 can be a human (gene ID: 55145), non-human primate (e.g., gene ID: 101926823), or rodent gene (e.g., gene ID: 73754, gene ID: 306547). In humans, mutations in the gene encoding THAP1 are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (as annotated by, for example, GenBank RefSeq accession numbers: NM_018105.3 and NM_199003.2) have been characterized.

[0194] TOR1A: When used herein, TOR1A refers to the gene encoding Torsin family 1 member A (also referred to as DQ2; AMC5; DYT1), and a is a member of the AAA family of adenosine triphosphatases (ATPases). In some embodiments, TOR1A can be a human (Gene ID: 1861), non-human primate (e.g., Gene ID: 102124758), or rodent gene (e.g., Gene ID: 30931, Gene ID: 266606). In humans, mutations in the gene encoding TOR1A are associated with the development of hereditary dystonia. In addition, multiple human transcript variants encoding different protein isoforms (e.g., as annotated by GenBank RefSeq accession number: NM_000113.3) have been characterized.

[0195] TPP1: When used herein, "TPP1" refers to the gene encoding tripeptidyl peptidase 1 (also known as GIG1, LPIC, SCAR7, and CLN2). The tripeptidyl peptidase 1 enzyme is involved in CLN2 Batten disease. Wild-type tripeptidyl peptidase 1 mediates the cleavage of the N-terminal tripeptide from the substrate. In CLN2 Batten disease, mutations in the tripeptidyl peptidase 1 enzyme severely reduce its enzymatic activity, leading to incomplete degradation and subsequent accumulation of proteins in the lysosome. The most frequent tripeptidyl peptidase 1 mutation is a single amino acid change. The inheritance of CLN2 Batten disease is autosomal recessive. In some embodiments, CLN2 can be a human (Gene ID: 1200), non-human primate (Gene ID: 709838), or rodent (Gene ID: 12751; Gene ID: 83534) gene.

[0196] Transferrin receptor: As used herein, the term "transferrin receptor" (also known as TFRC, CD71, p90, TFR, or TFR1) refers to an internalizing cell surface receptor that binds transferrin and facilitates iron uptake by endocytosis. In some embodiments, the transferrin receptor can be of human (NCBI Gene ID 7037), non-human primate (e.g., NCBI Gene ID 711568 or NCBI Gene ID 102136007), or rodent (e.g., NCBI Gene ID 22042) origin. Additionally, multiple human transcript variants encoding different isoforms of the receptor have been characterized (as annotated, for example, by GenBank RefSeq accession numbers: NP_001121620.1, NP_003225.2, NP_001300894.1, and NP_001300895.1).

[0197] TREM2: As used herein, TREM2 refers to the gene encoding Triggering Receptor Expressed on Myeloid cells 2, a protein involved in inflammation, synapse pruning, and neuron cell survival (also referred to as PLOSL2, Trem2a, Trem2b, and Trem2c). In the brain, TREM2 is expressed in microglial cells. In some embodiments, TREM2 can be a human (Gene ID: 54209), non-human primate (e.g., Gene ID: 719740), or rodent gene (e.g., Gene ID: 83433, Gene ID: 301227). In humans, mutations in the TREM2 gene are associated with the development of Alzheimer's disease. In some embodiments, mutations in TREM2 are associated with an increased risk of Alzheimer's disease. Genetic variants of TREM2 are also associated with an increased risk of multiple neurodegenerative diseases including frontotemporal dementia and Alzheimer's disease. See, e.g., Carmona, et al. “The role of TREM2 in Alzheimer’s disease and other neurodegenerative disorders” Lancet Neurology 17(8):721-730 (2018). Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, e.g., by GenBank RefSeq accession numbers: NM_018965.4 and NM_001271821.2) have been characterized.

[0198] UBE3A: As used herein, the term "UBE3A" refers to the gene encoding the ubiquitin protein ligase E3A, a protein involved in ubiquitination and proteolysis (also referred to as E6AP, ANCR, AS, EPVE6AP, HPVE6A, and PIX1). In some embodiments, UBE3A can be a human (Gene ID: 7337), non-human primate (e.g., Gene ID: 711270), or rodent gene (e.g., Gene ID: 22215, Gene ID: 361585). In humans, mutations in the UBE3A gene are associated with Angelman syndrome and autism spectrum disorder. Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_130838.4, NM_000462.5, and NM_130839.5) have been characterized.

[0199] UNC13A: When used herein, the term "UNC13A" refers to the gene encoding a member of the UNC13 family of proteins involved in calcium-triggered synaptic vesicle release, Unc-13 homolog A (also referred to as Munc13-1 and unc-13 homolog A (C. elegans)) (see, for example, J. S. Dittman “Unc13: a multifunctional synaptic marvel” Curr Opin Neurobiol. 57:17-25 (2019)). In some embodiments, UNC13A can be a human (Gene ID: 23025), non-human primate (for example, Gene ID: 720000, Gene ID: 102123626), or rodent gene (for example, Gene ID: 382018, Gene ID: 64829). Additionally, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_001080421.3, NM_001387021.1, NM_001387022.1, NM_001387023.1, XM_011527810.3, XM_011527811.3, XM_017026502.2, XM_054320277.1, XM_054320278.1, and XM_054320279.1) have been characterized. UNC13A contains a cryptic exon that promotes nonsense-mediated decay. Certain single nucleotide polymorphisms in UNC13A are associated with an increased risk of cryptic exon inclusion in UNC13A transcripts.Such polymorphisms in UNC13A are associated with neurodegenerative diseases such as ALS and frontotemporal dementia (see, for example, Brown, et al. “TDP-43 loss and ALS-risk SNPs drive mis-splicing and depletion of UNC13A” Nature 603:131-137 (2022); and Ma, et al. “TDP-43 represses cryptic exon inclusion in the FTD-ALS gene UNC13A” Nature 603:124-130 (2022)).

[0200] VLA-4: As used herein, the term “VLA-4” refers to the gene encoding the protein very late antigen 4, which is involved in cell adhesion and signaling (also referred to as ITGA4, CD49D, and IA4). In some embodiments, VLA-4 can be a human (Gene ID: 3676), non-human primate (e.g., Gene ID: 704745), or rodent gene (e.g., Gene ID: 16401, Gene ID: 311144). In humans, mutations in the VLA-4 gene are associated with retinitis pigmentosa 26, isolated macular dystrophy, and multiple sclerosis. In addition, multiple human transcript variants encoding different protein isoforms (as annotated, for example, by GenBank RefSeq accession numbers: NM_000885.6 and NM_001316312.2) have been characterized.

[0201] 2'-Modified Nucleosides: As used herein, the terms “2'-modified nucleoside” and “2'-modified ribonucleoside” are used interchangeably and refer to a nucleoside having a sugar moiety modified at the 2' position. In some embodiments, the 2'-modified nucleoside is a 2'-4' bicyclic nucleoside, where the 2' and 4' positions of the sugar are bridged (e.g., via a methylene, ethylene, or (S)-constrained ethyl bridge). In some embodiments, the 2'-modified nucleoside is an acyclic 2'-modified nucleoside, e.g., where the 2' position of the sugar moiety is substituted. Non-limiting examples of 2'-modified nucleosides include: 2'-deoxy, 2'-fluoro (2'-F), 2'-O-methyl (2'-O-Me), 2'-O-methoxyethyl (2'-MOE), 2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), 2'-O-dimethylaminoethyloxyethyl (2'-O-DMAEOE), 2'-O-N-methylacetamide (2'-O-NMA), locked nucleic acid (LNA, methylene-bridged nucleic acid), ethylene-bridged nucleic acid (ENA), and (S)-constrained ethyl-bridged nucleic acid (cEt). In some embodiments, the 2'-modified nucleosides described herein are high-affinity modified nucleosides, and oligonucleotides containing 2'-modified nucleosides have increased affinity for a target sequence relative to unmodified oligonucleotides. Examples of the structure of 2'-modified nucleosides are provided below: [Chemical Structure] These examples are shown with phosphate groups, but any internucleoside linkage is contemplated between 2'-modified nucleosides.

[0202] II. Complexes Combinations are provided herein that include a targeting agent, such as an antibody, covalently linked to a molecular payload. In some embodiments, the combination includes a central nervous system (CNS) targeting antibody covalently linked to an oligonucleotide. The combination can include an antibody that specifically binds to a single antigenic site or binds to at least two antigenic sites that can be present on the same or different antigens.

[0203] The combination can be used to modulate the activity or function of at least one gene, protein, and / or (by way of example, and) nucleic acid in CNS cells or to alleviate the symptoms of a CNS disease disorder. In some embodiments, the molecular payload present in the combination is responsible for the regulation of genes, proteins, and / or (by way of example, and) nucleic acids. The molecular payload can be a small molecule, protein, nucleic acid, oligonucleotide, or any molecular entity that can modulate the activity or function of a gene, protein, and / or (by way of example, and) nucleic acid in a cell.

[0204] In some embodiments, the CNS targeting agent of the combination described herein includes an anti-transferrin receptor 1 (TfR1) antibody covalently linked to a molecular payload, such as an oligonucleotide, polypeptide, small molecule, or gene therapy payload.

[0205] antibody In some embodiments, the combination described herein includes an antibody that binds to human transferrin receptor 1 (TfR1). An exemplary human TfR1 amino acid sequence corresponding to NCBI sequence NP_003225.2 (transferrin receptor protein 1 isoform 1, Homo sapiens) is as follows: MMDQARSAFSNLFGGEPLSYTRFSLARQVDGDNSHVEMKLAVDEEENADNNTKANVTKPKRCSGSICYGTIAVIVFFLIGFMIGYLGYCKGVEPKTECERLAGTESPVREEPGEDFPAARRLYWDDLKRKLSEKLDSTDFTGTIKLLNENSYVPREAGSQKDENLALYVENQFREFKLSKVWRDQHFVKIQVKDSAQNSVIIVDKNGRLVYLVENPGGYVAYSKAATVTGKLVHANFGTKKDFEDLYTPVNGSIVIVRAGKITFAEKVANAESLNAIGVLIYMDQTKFPIVNAELSFFGHAHLGTGDPYTPGFPSFNHTQFPPSRSSGLPNIPVQTISRAAAEKLFGNMEGDCPSDWKTDSTCRMVTSESKNVKLTVSNVLKEIKILNIFGVIKGFVEPDHYVVVGAQRDAWGPGAAKSGVGTALLLKLAQMFSDMVLKDGFQPSRSIIFASWSAGDFGSVGATEWLEGYLSSLHLKAFTYINLDKAVLGTSNFKVSASPLLYTLIEKTMQNVKHPVTGQFLYQDSNWASKVEKLTLDNAAFPFLAYSGIPAVSFCFCEDTDYPYLGTTMDTYKELIERIPELNKVARAAAEVAGQFVIKLTHDVELNLDYERYNSQLLSFVRDLNQYRADIKEMGLSLQWLYSARGDFFRATSRLTTDFGNAEKTDRFVMKKLNDRVMRVEYHFLSPYVSPKESPFRHVFWGSGSHTLPALLENLKLRKQNNGAFNETLFRNQLALATWTIQGAANALSGDVWDIDNEF(SEQ ID NO: 21)

[0206] Table 2 provides examples of sequences of anti-TfR1 antibodies useful for the complexes provided herein.

Table 2-1

Table 2-2

[0207] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises heavy chain complementarity determining region 1 (CDR-H1) of SEQ ID NO: 1 (according to the IMGT definition system), heavy chain complementarity determining region 2 (CDR-H2) of SEQ ID NO: 2 (according to the IMGT definition system), heavy chain complementarity determining region 3 (CDR-H3) of SEQ ID NO: 3 (according to the IMGT definition system), light chain complementarity determining region 1 (CDR-L1) of SEQ ID NO: 4 (according to the IMGT definition system), light chain complementarity determining region 2 (CDR-L2) of SEQ ID NO: 5 (according to the IMGT definition system), and light chain complementarity determining region 3 (CDR-L3) of SEQ ID NO: 6 (according to the IMGT definition system).

[0208] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises heavy chain complementarity determining region 1 (CDR-H1) of SEQ ID NO: 7 (according to the Kabat definition system), heavy chain complementarity determining region 2 (CDR-H2) of SEQ ID NO: 8 (according to the Kabat definition system), heavy chain complementarity determining region 3 (CDR-H3) of SEQ ID NO: 9 (according to the Kabat definition system), light chain complementarity determining region 1 (CDR-L1) of SEQ ID NO: 10 (according to the Kabat definition system), light chain complementarity determining region 2 (CDR-L2) of SEQ ID NO: 11 (according to the Kabat definition system), and light chain complementarity determining region 3 (CDR-L3) of SEQ ID NO: 6 (according to the Kabat definition system).

[0209] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises heavy chain complementarity determining region 1 (CDR-H1) of SEQ ID NO: 12 (according to the Chothia definition system), heavy chain complementarity determining region 2 (CDR-H2) of SEQ ID NO: 13 (according to the Chothia definition system), heavy chain complementarity determining region 3 (CDR-H3) of SEQ ID NO: 14 (according to the Chothia definition system), light chain complementarity determining region 1 (CDR-L1) of SEQ ID NO: 15 (according to the Chothia definition system), light chain complementarity determining region 2 (CDR-L2) of SEQ ID NO: 5 (according to the Chothia definition system), and light chain complementarity determining region 3 (CDR-L3) of SEQ ID NO: 16 (according to the Chothia definition system).

[0210] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises a variable heavy chain region (VH) that contains at most 25 amino acid variations (e.g., at most 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) in the framework region as compared to the VH comprising the amino acid sequence of SEQ ID NO: 17. Alternatively or additionally (e.g., additionally), the anti-TfR1 antibody of the present disclosure comprises a variable light chain region (VL) that contains at most 25 amino acid variations (e.g., at most 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid variation) in the framework region as compared to the VL comprising the amino acid sequence of SEQ ID NO: 18.

[0211] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises a VH that comprises an amino acid sequence that is at least 75% (e.g., 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VH comprising the amino acid sequence of SEQ ID NO: 17 in the framework region. Alternatively or additionally (e.g., additionally), in some embodiments, the anti-TfR1 antibody of the present disclosure comprises a VL that comprises an amino acid sequence that is at least 75% (e.g., 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the VL comprising the amino acid sequence of SEQ ID NO: 18 in the framework region.

[0212] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises a VH comprising the amino acid sequence of SEQ ID NO: 17. Alternatively or additionally (e.g., additionally), in some embodiments, the anti-TfR1 antibody of the present disclosure comprises a VL comprising the amino acid sequence of SEQ ID NO: 18.

[0213] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises a heavy chain comprising an amino acid sequence that is at least 75% (by way of example, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition (by way of addition), the anti-TfR1 antibody of the present disclosure comprises a light chain comprising an amino acid sequence that is at least 75% (by way of example, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the amino acid sequence of SEQ ID NO: 20. In some embodiments, the anti-TfR1 antibody of the present disclosure is a Fab comprising a heavy chain comprising an amino acid sequence that is at least 75% (by way of example, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition (by way of addition), the anti-TfR1 antibody of the present disclosure is a Fab comprising a light chain comprising an amino acid sequence that is at least 75% (by way of example, 75%, 80%, 85%, 90%, 95%, 98%, or 99%) identical to the amino acid sequence of SEQ ID NO: 20.

[0214] In some embodiments, the anti-TfR1 antibody of the present disclosure comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition (by way of addition), the anti-TfR1 antibody of the present disclosure comprises a light chain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the anti-TfR1 antibody of the present disclosure is a Fab comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition (by way of addition), the anti-TfR1 antibody of the present disclosure is a Fab comprising a light chain comprising the amino acid sequence of SEQ ID NO: 20.

[0215] In some embodiments, the anti-TfR1 antibodies provided herein may have one or more post-translational modifications. In some embodiments, N-terminal cyclization, also referred to as pyroglutamic acid formation (pyroGlu), may occur on the antibody at the N-terminal glutamate (Glu) and / or glutamine (Gln) residues during production. Thus, it should be understood that antibodies designated as having a sequence containing an N-terminal glutamate or glutamine residue include antibodies that have undergone pyroglutamic acid formation resulting from post-translational modification. In some embodiments, pyroglutamic acid formation occurs in the heavy chain sequence. In some embodiments, pyroglutamic acid formation occurs in the light chain sequence.

[0216] Molecular payload Some aspects of the present disclosure provide, by way of example, a molecular payload for modulating biological outcomes such as, by way of example, transcription of DNA sequences, processing of pre-mRNA transcripts, stability of pre-mRNA or mRNA transcripts, protein expression (e.g., translation of mRNA), or protein activity, which can be linked to the anti-TfR1 antibodies described herein (e.g., the anti-TfR1 antibodies in Table 2). In some embodiments, such molecular payloads are targeted to CNS cells, for example, via specific binding to nucleic acids or proteins within or on CNS cells after delivery of the linked anti-TfR1 antibody to the CNS cells. It should be understood that various types of molecular payloads can be used in accordance with the present disclosure. For example, the molecular payload can include, or consist of, an oligonucleotide (e.g., an antisense oligonucleotide or an RNA interference oligonucleotide), a polypeptide (e.g., a peptide, protein, or antibody that binds to a nucleic acid or protein in CNS cells), a small molecule (e.g., a small molecule that modulates the function of a nucleic acid or protein in CNS cells), or a gene therapy payload (e.g., a nucleic acid encoding a polypeptide having biological activity in CNS cells).

[0217] In some embodiments, the molecular payload is an oligonucleotide comprising a strand having a region of complementarity to a gene (e.g., a gene transcript) provided in Table 3. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7]

[0218] In some embodiments, the molecular payload is an oligonucleotide comprising a strand having a region of complementarity to a gene (e.g., a gene transcript) provided in Table 4. [Table 4-1] [Table 4-2] [Table 4-3]

[0219] Oligonucleotide Payload In some embodiments, the oligonucleotide is useful for treating various CNS diseases and disorders. For example, the oligonucleotide can be useful for regulating the expression or activity of various genes involved in CNS diseases and disorders by, for example, regulating gene transcription, regulating the stability of mRNA molecules encoded by the gene, regulating the translation of mRNA molecules encoded by the gene, and regulating the splicing of pre-mRNA transcripts encoded by the gene. The oligonucleotide can be used to treat various CNS diseases and disorders, for example, by facilitating the delivery of the oligonucleotide into cells of the CNS. In some embodiments, the oligonucleotides disclosed herein can be delivered into cells of the CNS using the complexes disclosed herein (e.g., an anti-TfR1 antibody complex comprising an oligonucleotide).

[0220] In some embodiments, the oligonucleotide is useful for treating a neuromuscular disease or disorder (e.g., Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy); amyotrophic lateral sclerosis; Parkinson's disease; Huntington's disease; Alzheimer's disease; epilepsy; a pain disorder; a glycogen synthesis disorder; neurodegeneration; small fiber neuropathy; a phenotype related to nociception; Alexander disease; Angelman syndrome; an autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis.

[0221] In some embodiments, the oligonucleotide is useful for treating essential tremor and / or hereditary dystonia.

[0222] In some embodiments, the oligonucleotide is useful for the regulation of one or more genes associated with CNS diseases or disorders. In some embodiments, one or more genes associated with CNS diseases or disorders are DMPK, DMD, SMN, FXN, SOD1, C9orf72, ATXN2, FUS, LRRK2, SNCA, HTT, MSH3, TREM2, APOE, MAPT, APP, GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, SCN9A, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, or PCDH19. In some embodiments, one or more genes associated with CNS diseases or disorders are TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1. In some embodiments, one or more genes associated with CNS diseases or disorders are PIKFYVE, SYF2, UNC13A, ATXN1, ATXN3, GRN, GRIN2A, TPP1, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2.

[0223] In some embodiments, any 5' or 3' nucleoside (such as the terminal nucleoside) of any one of the oligonucleotides described herein is optionally conjugated to an amine group via a spacer. In some embodiments, the spacer includes an aliphatic moiety. In some embodiments, the spacer includes a polyethylene glycol moiety. In some embodiments, a phosphodiester linkage is present between the spacer and the 5' or 3' nucleoside of the oligonucleotide. In some embodiments, any 5' or 3' nucleoside (such as the terminal nucleoside) of any one of the oligonucleotides described herein is conjugated to a spacer, which is a substituted or unsubstituted aliphatic, substituted or unsubstituted heteroaliphatic, substituted or unsubstituted carbocyclylene, substituted or unsubstituted heterocyclylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, -O-, -N(RA )-, -S-, -C(=O)-, -C(=O)O-, -C(=O)NR A -, -NR A C(=O)-, -NR A C(=O)R A -, -C(=O)R A -, -NR A C(=O)O-, -NR A C(=O)N(R A )-, -OC(=O)-, -OC(=O)O-, -OC(=O)N(R A )-, -S(O)2NR A -, -NR A S(O)2-, or a combination thereof; each R A is independently hydrogen or substituted or unsubstituted alkyl. In certain embodiments, the spacer is substituted or unsubstituted alkylene, substituted or unsubstituted heterocyclylene, substituted or unsubstituted heteroarylene, -O-, -N(R A )-, or -C(=O)N(R A )2, or a combination thereof.

[0224] In some embodiments, the 5' or 3' nucleoside of any one of the oligonucleotides described herein is conjugated to a compound of the formula -NH2-(CH2) n -, wherein n is an integer from 1 to 12. In some embodiments, n is 6, 7, 8, 9, 10, 11, or 12. In some embodiments, a phosphodiester linkage is present between a compound of the formula NH2-(CH2) n - and the 5' or 3' nucleoside of the oligonucleotide. In some embodiments, a compound of the formula NH2-(CH2)6- is conjugated to the oligonucleotide via a reaction between 6-amino-1-hexanol (NH2-(CH2)6-OH) and the 5'-phosphate of the oligonucleotide.

[0225] In some embodiments, the oligonucleotide is conjugated to a targeting agent, such as a CNS targeting agent such as an anti-TfR1 antibody, via, for example, an amine group.

[0226] a. Oligonucleotide Size / Sequence Oligonucleotides can be of various different lengths, depending, for example, on the format. In some embodiments, the oligonucleotide is 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 75, or more nucleotides in length. In some embodiments, the oligonucleotide is 8 - 50 nucleotides in length, 8 - 40 nucleotides in length, 8 - 30 nucleotides in length, 10 - 15 nucleotides in length, 10 - 20 nucleotides in length, 15 - 25 nucleotides in length, 21 - 23 nucleotides in length, 20 - 25 nucleotides in length, etc.

[0227] In some embodiments, the nucleic acid sequence of the oligonucleotide for the purposes of the present disclosure is "complementary" to the target nucleic acid when it is capable of specifically hybridizing to the target nucleic acid. In some embodiments, an oligonucleotide hybridizing to a target nucleic acid (e.g., a transcript provided in Table 3, provided by any one of SEQ ID NOs: 392 - 702) results in modulation of the activity or expression of the target (e.g., decreased mRNA translation, modulated pre-mRNA splicing, exon skipping, target mRNA degradation, etc.). In some embodiments, an oligonucleotide hybridizing to a target nucleic acid (e.g., a transcript provided in Table 4, provided by any one of SEQ ID NOs: 705 - 803) results in modulation of the activity or expression of the target (e.g., decreased mRNA translation, modulated pre-mRNA splicing, exon skipping, target mRNA degradation, etc.). In some embodiments, an oligonucleotide hybridizing to a target nucleic acid (e.g., a transcript provided in Table 3 or Table 4, provided by any one of SEQ ID NOs: 143 - 148, 167 - 169, 810 - 875, and 1059 - 1068) results in modulation of the activity or expression of the target (e.g., decreased mRNA translation, modulated pre-mRNA splicing, exon skipping, target mRNA degradation, etc.). In some embodiments, an oligonucleotide hybridizing to a target nucleic acid results in an increase in the activity or expression of the target (e.g., increased mRNA translation such as the wild-type form of the mRNA; modulated pre-mRNA splicing; exon skipping; target mRNA stabilization, etc.). In some embodiments, the nucleic acid sequence of the oligonucleotide has a sufficient degree of complementarity to its target nucleic acid such that under conditions where avoidance of non-specific binding is desired, e.g., under physiological conditions, it does not hybridize to non-target sequences.Thus, in some embodiments, the oligonucleotide can be at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complementary to consecutive nucleotides of the target nucleic acid. In some embodiments, the complementary nucleotide sequence need not be 100% complementary to that of its target in order to be capable of specifically hybridizing or being specific to the target nucleic acid. In certain embodiments, the oligonucleotide contains one or more mismatched nucleobases relative to the target nucleic acid. In certain embodiments, the activity with respect to the target is reduced by such mismatches, while the activity with respect to non-targets is reduced by a greater amount (i.e., the selectivity for the target nucleic acid is increased and the off-target effect is decreased).

[0228] In some embodiments, the oligonucleotide contains a region of complementarity to a target nucleic acid that ranges in length from 8 to 15, 8 to 30, 8 to 40, or 10 to 50, or 5 to 50, 15 to 20, 20 to 25, or 5 to 40 nucleotides. In some embodiments, the region of complementarity of the oligonucleotide to the target nucleic acid is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 nucleotides in length. In some embodiments, the region of complementarity is complementary to at least 8 consecutive nucleotides of the target nucleic acid. In some embodiments, the oligonucleotide can contain 1, 2 or 3 base mismatches compared to a portion of consecutive nucleotides of the target nucleic acid. In some embodiments, the oligonucleotide can have up to 3 mismatches in 15 bases, or up to 2 mismatches in 10 bases.

[0229] In some embodiments, the oligonucleotide is (e.g., at least 85%, at least 90%, at least 95%, or 100%) complementary to any one of the target sequences of the oligonucleotides described herein (e.g., the oligonucleotides listed in Tables 5-19). In some embodiments, the oligonucleotide is (e.g., at least 85%, at least 90%, at least 95%, or 100%) complementary to the target sequences provided herein (e.g., the transcripts listed in Table 3 provided by any one of SEQ ID NOs: 392-702). In some embodiments, the oligonucleotide is (e.g., at least 85%, at least 90%, at least 95%, or 100%) complementary to the target sequences provided herein (e.g., the transcripts listed in Table 4 provided by any one of SEQ ID NOs: 705-803). In some embodiments, the oligonucleotide is (e.g., at least 85%, at least 90%, at least 95%, or 100%) complementary to the target sequences provided herein (e.g., the transcripts listed in Table 3 or Table 4 provided by any one of SEQ ID NOs: 143-148, 167-169, 810-875, and 1059-1068).

[0230] In some embodiments, oligonucleotides useful for targeting the transcripts provided herein include sequences comprising at least 8 (e.g., 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more) consecutive nucleobases of the oligonucleotide sequences provided herein (e.g., the oligonucleotide sequences listed in any one of Tables 5-19). In some embodiments, oligonucleotides useful for targeting the transcripts provided herein include sequences comprising a region of complementarity of at least 8 (e.g., 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, or more) consecutive nucleobases that are complementary to a target sequence of the oligonucleotide sequences provided herein (e.g., the oligonucleotide sequences listed in any one of Tables 5-19).

[0231] In some embodiments, it should be understood that methylation of the nucleobase uracil at the C5 position forms thymine. Therefore, in some embodiments, a nucleotide or nucleoside having C5-methylated uracil (or 5-methyl-uracil) can be equivalently identified as a thymine nucleotide or nucleoside.

[0232] In some embodiments, any one or more of the thymine bases (T) in any one of the oligonucleotides provided herein (e.g., the oligonucleotides listed in any one of Tables 5-19) can independently and optionally be uracil bases (U), and / or any one or more of the Us in the oligonucleotides provided herein can independently and optionally be Ts.

[0233] b. Oligonucleotide modification: The oligonucleotides described herein can be modified. By way of example, it can include modified sugar moieties, modified internucleoside linkages, modified nucleotides or nucleosides, and / or combinations thereof (by way of example, and). In addition, in some embodiments, the oligonucleotide can exhibit one or more of the following properties: not mediating alternative splicing; not being immunostimulatory; being nuclease resistant; having improved cellular uptake compared to unmodified oligonucleotides; not being toxic to cells or mammals; having improved endosomal escape inside cells; minimizing TLR stimulation; or avoiding pattern recognition receptors. Any of the modified chemistries or formats of the oligonucleotides described herein can be combined with each other. For example, 1, 2, 3, 4, 5, or more different types of modifications can be incorporated within the same oligonucleotide.

[0234] In some embodiments, certain nucleotide or nucleoside modifications that make the oligonucleotides into which they are incorporated more resistant to nuclease digestion than natural state oligodeoxynucleotide or oligoribonucleotide molecules can be used; these modified oligonucleotides survive intact for a longer time than unmodified oligonucleotides. Specific examples of modified oligonucleotides include those with modified backbones, such as modified internucleoside linkages, such as phosphorothioates, phosphotriesters, methylphosphonates, short chain alkyl or cycloalkyl sugar linkages, or short chain heteroatom or heterocyclic sugar linkages. Thus, the oligonucleotides of the present disclosure can be stabilized against nuclease degradation, by way of example, by the incorporation of modifications, by way of example, nucleotide or nucleoside modifications.

[0235] In some embodiments, the oligonucleotide can be up to 50 or up to 100 nucleotides in length, wherein 2 to 10, 2 to 15, 2 to 16, 2 to 17, 2 to 18, 2 to 19, 2 to 20, 2 to 25, 2 to 30, 2 to 40, 2 to 45, or more nucleotides or nucleosides of the oligonucleotide are modified nucleotides / nucleosides. The oligonucleotide can be 8 to 30 nucleotides in length, wherein 2 to 10, 2 to 15, 2 to 16, 2 to 17, 2 to 18, 2 to 19, 2 to 20, 2 to 25, 2 to 30 nucleotides or nucleosides of the oligonucleotide are modified nucleotides / nucleosides. The oligonucleotide can be 8 to 15 nucleotides in length, wherein 2 to 4, 2 to 5, 2 to 6, 2 to 7, 2 to 8, 2 to 9, 2 to 10, 2 to 11, 2 to 12, 2 to 13, 2 to 14 nucleotides or nucleosides of the oligonucleotide are modified nucleotides / nucleosides. Optionally, the oligonucleotide can be modified at every nucleotide or nucleoside, except for 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides / nucleosides. Oligonucleotide modifications are further described herein.

[0236] c. Modified nucleoside In some embodiments, the oligonucleotides described herein include at least one nucleoside having a modified 2'-position of the sugar. In some embodiments, the oligonucleotide includes at least one 2'-modified nucleoside. In some embodiments, all of the nucleosides on the oligonucleotide are 2'-modified nucleosides.

[0237] In some embodiments, the oligonucleotides described herein include one or more non-bicyclic 2'-modified nucleosides, such as 2'-deoxy, 2'-fluoro (2'-F), 2'-O-methyl (2'-O-Me), 2'-O-methoxyethyl (2'-MOE), 2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), 2'-O-dimethylaminoethyloxyethyl (2'-O-DMAEOE), or 2'-O-N-methylacetamide (2'-O-NMA) modified nucleosides.

[0238] In some embodiments, the oligonucleotides described herein include one or more 2'-4' bicyclic nucleosides, wherein the ribose ring includes a bridging moiety connecting two atoms on the ring, and by way of example, connects the 2'-O atom to the 4'-C atom via a methylene (LNA) bridge, an ethylene (ENA) bridge, or an (S)-constrained ethyl (cEt) bridge. Examples of LNA are described in International Patent Application Publication WO / 2008 / 043753 entitled "RNA Antagonist Compounds For The Modulation Of PCSK9", published Apr. 17, 2008. The contents thereof are incorporated herein by reference in their entirety. Examples of ENA are provided in International Patent Publication No. WO 2005 / 042777, published May 12, 2005, entitled "APP / ENA Antisense"; Morita et al., Nucleic Acid Res., Suppl 1:241-242, 2001; Surono et al., Hum.Gene Ther., 15:749-757, 2004; Koizumi, Curr.Opin. Mol. Ther., 8:144-149, 2006, and Horie et al., Nucleic Acids Symp.Ser (Oxf), 49:171-172, 2005; the disclosures of which are incorporated herein by reference in their entirety. Examples of cEt are provided in U.S. Patents 7,101,993; 7,399,845, and 7,569,686, each of which is incorporated herein by reference in its entirety.

[0239] In some embodiments, the oligonucleotide comprises modified nucleosides disclosed in one of the following U.S. patents or patent application publications: U.S. Patent 7,399,845, titled "6-Modified Bicyclic Nucleic Acid Analogs," issued July 15, 2008; U.S. Patent 7,741,457, titled "6-Modified Bicyclic Nucleic Acid Analogs," issued June 22, 2010; U.S. Patent 8,022,193, titled "6-Modified Bicyclic Nucleic Acid Analogs," issued September 20, 2011; U.S. Patent 7,569,686, titled "Compounds And Methods For Synthesis Of Bicyclic Nucleic Acid Analogs," issued August 4, 2009; U.S. Patent 7,335,765, titled "Novel Nucleoside And Oligonucleotide Analogues," issued February 26, 2008; U.S. Patent 7,314,923, titled "Novel Nucleoside And Oligonucleotide Analogues," issued January 1, 2008; U.S. Patent 7,816,333, titled "Oligonucleotide Analogues And Methods Utilizing The Same," issued October 19, 2010, and U.S. Publication No. 2011 / 0009471, titled "Oligonucleotide Analogues And Methods Utilizing The Same," published February 17, 2015, now U.S. Patent 8,957,201. The entire contents of each of these are hereby incorporated by reference herein for all purposes.

[0240] In some embodiments, the oligonucleotide comprises at least one modified nucleoside, which results in an increase in the Tm of the oligonucleotide in the range of 1 °C, 2 °C, 3 °C, 4 °C, or 5 °C compared to an oligonucleotide having no modified nucleosides. The oligonucleotide can have a plurality of modified nucleosides that result in a total increase in the Tm of the oligonucleotide in the range of 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, 10 °C, 15 °C, 20 °C, 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, or more compared to an oligonucleotide having no modified nucleosides.

[0241] The oligonucleotide can comprise a mix of different types of nucleosides. For example, the oligonucleotide can comprise a mix of 2'-deoxyribonucleosides or ribonucleosides and 2'-fluoro-modified nucleosides. The oligonucleotide can comprise a mix of deoxyribonucleosides or ribonucleosides and 2'-O-Me modified nucleosides. The oligonucleotide can comprise a mix of 2'-fluoro-modified nucleosides and 2'-O-Me modified nucleosides. The oligonucleotide can comprise a mix of 2'-4'-bicyclic nucleosides and 2'-MOE, 2'-fluoro, or 2'-O-Me modified nucleosides. The oligonucleotide can comprise a mix of non-bicyclic 2'-modified nucleosides (e.g., 2'-MOE, 2'-fluoro, or 2'-O-Me) and 2'-4'-bicyclic nucleosides (e.g., LNA, ENA, cEt).

[0242] Oligonucleotides can contain alternating types of nucleosides. For example, an oligonucleotide can contain alternating 2'-deoxyribonucleosides or ribonucleosides and 2'-fluoro-modified nucleosides. An oligonucleotide can contain alternating deoxyribonucleosides or ribonucleosides and 2'-O-Me-modified nucleosides. An oligonucleotide can contain alternating 2'-fluoro-modified nucleosides and 2'-O-Me-modified nucleosides. An oligonucleotide can contain alternating 2'-4'-bicyclic nucleosides and 2'-MOE, 2'-fluoro, or 2'-O-Me-modified nucleosides. An oligonucleotide can contain alternating acyclic 2'-modified nucleosides (by way of example, 2'-MOE, 2'-fluoro, or 2'-O-Me) and 2'-4'-bicyclic nucleosides (by way of example, LNA, ENA, cEt).

[0243] In some embodiments, the oligonucleotides described herein contain 5'-vinylphosphonate modifications, one or more abasic residues, and / or one or more inverted abasic residues.

[0244] d. Inter-nucleoside Linkage / Backbone In some embodiments, an oligonucleotide can contain phosphorothioate or other modified inter-nucleoside linkages. In some embodiments, an oligonucleotide contains phosphorothioate inter-nucleoside linkages. In some embodiments, an oligonucleotide contains phosphorothioate inter-nucleoside linkages between at least two nucleosides. In some embodiments, an oligonucleotide contains phosphorothioate inter-nucleoside linkages between all nucleosides. For example, in some embodiments, an oligonucleotide contains a modified inter-nucleoside linkage in the first, second, and / or (by way of example, and) third inter-nucleoside linkages at the 5' or 3' end of the nucleotide sequence.

[0245] Phosphorus-containing linkages that can be used include phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl phosphotriesters, methyl and other alkyl phosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3'-aminophosphoramidates, as well as aminoalkyl phosphoramidates, thionophosphoramidates, thionoalkyl phosphonates, thionoalkyl phosphotriesters, and boranophosphates having normal 3'-5' linkages, 2'-5' linkage analogs thereof, and those having an inverted polarity where adjacent pairs of nucleoside units are linked from 3'-5' to 5'-3' or from 2'-5' to 5'-2', but are not limited thereto; see U.S. Patent Nos. 3,687,808; 4,469,863; 4,476,301; 5,023,243; 5,177,196; 5,188,897; 5,264,423; 5,276,019; 5,278,302; 5,286,717; 5,321,131; 5,399,676; 5,405,939; 5,453,496; 5,455,233; 5,466,677; 5,476,925; 5,519,126; 5,536,821; 5,541,306; 5,550,111; 5,563,253; 5,571,799; 5,587,361; and 5,625,050.

[0246] In some embodiments, the oligonucleotide can have a heteroatom backbone, such as a methylene(methylimino) or MMI backbone; an amide backbone (see De Mesmaeker et al. Ace. Chem. Res. 1995, 28:366 - 374); a morpholino backbone (see Summerton and Weller, U.S. Pat. No. 5,034,506); or a peptide nucleic acid (PNA) backbone (wherein the phosphodiester backbone of the oligonucleotide is replaced by a polyamide backbone and the nucleotides are attached directly or indirectly to the azanitrogen atoms of the polyamide backbone. See Nielsen et al., Science 1991, 254, 1497).

[0247] e. Stereospecific oligonucleotides In some embodiments, the internucleotidic phosphorus atoms of the oligonucleotide are chiral and the properties of the oligonucleotide are modulated based on the configuration of the chiral phosphorus atoms. In some embodiments, suitable methods can be used to synthesize P-chiral oligonucleotide analogs in a stereocontrolled manner (for example, as described in Oka N, Wada T, Stereocontrolled synthesis of oligonucleotide analogs containing chiral internucleotidic phosphorus atoms. Chem Soc Rev. 2011 Dec;40(12):5829-43). In some embodiments, phosphorothioate-containing oligonucleotides are provided that contain nucleoside units linked together by phosphorothioate sugar linkages that are substantially all Sp or substantially all Rp. In some embodiments, such phosphorothioate oligonucleotides having substantially enantiopure sugar linkages are prepared by enzymatic or chemical synthesis, as described, for example, in U.S. Patent 5,587,261, issued December 12, 1996. The contents of which are incorporated herein by reference in their entirety. In some embodiments, chirally controlled oligonucleotides provide a selective cleavage pattern of a target nucleic acid. For example, in some embodiments, a chirally controlled oligonucleotide provides cleavage at a single site within a complementary sequence of a nucleic acid. For example, as described in U.S. Patent Application Publication 20170037399 A1, entitled "CHIRAL DESIGN," published February 2, 2017, the contents of which are incorporated herein by reference in their entirety.

[0248] f. Morpholino In some embodiments, the oligonucleotide can be a morpholino-based compound. Morpholino-based oligomeric compounds are described in Dwaine A. Braasch and David R. Corey, Biochemistry, 2002, 41(14), 4503-4510); Genesis, volume 30, issue 3, 2001; Heasman, J., Dev. Biol., 2002, 243, 209-214; Nasevicius et al., Nat. Genet., 2000, 26, 216-220; Lacerra et al., Proc. Natl. Acad. Sci., 2000, 97, 9591-9596; and U.S. Pat. No. 5,034,506, issued Jul. 23, 1991. In some embodiments, the morpholino-based oligomeric compound is a phosphorodiamidate morpholino oligomer (PMO) (as described, for example, in Iverson, Curr. Opin. Mol. Ther., 3:235-238, 2001; and Wang et al., J. Gene Med., 12:354-364, 2010; the disclosures of which are incorporated herein by reference in their entirety).

[0249] g. Peptide nucleic acid (PNA) In some embodiments, both the sugar of the nucleotide unit and the internucleoside linkage (backbone) of the oligonucleotide are replaced by novel groups. In some embodiments, the base units are maintained for hybridization to a suitable nucleic acid target compound. One such oligomeric compound, an oligonucleotide mimetic that has been shown to have excellent hybridization properties, is called a peptide nucleic acid (PNA). In a PNA compound, the sugar backbone of the oligonucleotide is replaced by an amide-containing backbone, such as an aminoethylglycine backbone. The nucleobases are retained and are attached directly or indirectly to the azanitrogen atoms of the amide portions of the backbone. Representative publications reporting the preparation of PNA compounds include, but are not limited to, U.S. Patent Nos. 5,539,082; 5,714,331; and 5,719,262. Each of these is incorporated herein by reference. Further teachings of PNA compounds can be found in Nielsen et al., Science, 1991, 254, 1497-1500.

[0250] h. Mixmer In some embodiments, the oligonucleotides described herein can be mixmers or can contain mixmer sequence patterns. Generally, mixmers are oligonucleotides that contain both naturally occurring and non-naturally occurring nucleosides, or typically contain two different types of non-naturally occurring nucleosides in an alternating pattern. Mixmers generally have higher binding affinities than unmodified oligonucleotides and can be used, for example, to specifically bind to a target molecule to block a binding site on the target molecule. Generally, mixmers do not recruit RNase to the target molecule and thus do not promote cleavage of the target molecule. Such oligonucleotides that cannot recruit RNase H have been described. See, for example, WO2007 / 112754 or WO2007 / 112753.

[0251] In some embodiments, the mixmer comprises, or consists of, a repeating pattern of nucleoside analogs and naturally occurring nucleosides, or a first type of nucleoside analog and a second type of nucleoside analog. However, the mixmer need not comprise a repeating pattern, and instead may comprise any arrangement of modified nucleosides and naturally occurring nucleosides, or any arrangement of a first type of modified nucleoside and a second type of modified nucleoside. The repeating pattern can be, for example, where every second or every third nucleoside is a modified nucleoside such as an LNA, and the remaining nucleosides are naturally occurring nucleosides such as DNA, or 2'-substituted nucleoside analogs such as 2'-MOE or 2'-fluoro analogs, or any other modified nucleoside described herein. It is recognized that repeating patterns of modified nucleosides such as LNA units can be combined with modified nucleosides at fixed positions, such as at the 5' or 3' termini, by way of example.

[0252] In some embodiments, the mixmer does not contain a region of more than 5, more than 4, more than 3, or more than 2 consecutive natural nucleosides, such as DNA nucleosides. In some embodiments, the mixmer comprises a region consisting of at least, at least 2 consecutive modified nucleosides, such as at least 2 consecutive LNAs. In some embodiments, the mixmer comprises a region consisting of at least, at least 3 consecutive modified nucleoside units, such as at least 3 consecutive LNAs.

[0253] In some embodiments, the mixmer does not contain a region of more than 7, more than 6, more than 5, more than 4, more than 3, or more than 2 consecutive nucleoside analogs, such as LNAs. In some embodiments, the LNA units can be replaced by other nucleoside analogs such as those referenced herein.

[0254] Mixmers can be designed to include, in non-limiting examples, mixtures of affinity-enhancing modified nucleosides such as LNA nucleosides and 2'-O-Me nucleosides. In some embodiments, the mixmer includes modified internucleoside linkages (e.g., phosphorothioate internucleoside linkages or other linkages) between at least two, at least three, at least four, at least five, or more nucleosides.

[0255] Mixmers can be produced using any suitable method. Representative U.S. patents, U.S. patent publications, and PCT publications that teach the preparation of mixmers include U.S. Patent Publication Nos. US20060128646, US20090209748, US20090298916, US20110077288, and US20120322851, as well as U.S. Patent No. 7687617.

[0256] In some embodiments, the mixmer includes one or more morpholino nucleosides. For example, in some embodiments, the mixmer can include morpholino nucleosides mixed (e.g., in an alternating fashion) with one or more other nucleosides (e.g., DNA, RNA nucleosides) or modified nucleosides (e.g., LNA, 2'-O-Me nucleosides).

[0257] In some embodiments, mixmers are useful for splice correction or exon skipping. For example, as described in Touznik A., et al., LNA / DNA mixmer-based antisense oligonucleotides correct alternative splicing of the SMN2 gene and restore SMN protein expression in type 1 SMA fibroblasts Scientific Reports, volume 7, Article number: 3672 (2017), Chen S. et al., Synthesis of a Morpholino Nucleic Acid (MNA)-Uridine Phosphoramidite, and Exon Skipping Using MNA / 2’-O-Methyl Mixmer Antisense Oligonucleotide, Molecules 2016, 21, 1582. The contents of each of these are hereby incorporated by reference.

[0258] i. Multimer In some embodiments, the molecular payload can include multimers (such as concatemers) of two or more oligonucleotides connected by linkers. In this way, in some embodiments, the oligonucleotide payload of the complex can be increased over the available ligation sites on the targeting agent (such as the available thiol sites on an antibody), or otherwise tuned to achieve a particular payload content. The oligonucleotides in the multimer can be the same or different (such as targeting different genes, or different sites on the same gene, or their products).

[0259] In some embodiments, the multimer comprises two or more oligonucleotides linked together by a cleavable linker. However, in some embodiments, the multimer comprises two or more oligonucleotides linked together by a non-cleavable linker. In some embodiments, the multimer comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, or more oligonucleotides linked together. In some embodiments, the multimer comprises 2 to 5, 2 to 10, or 4 to 20 oligonucleotides linked together.

[0260] In some embodiments, the multimer comprises two or more oligonucleotides linked end-to-end (in a linear arrangement). In some embodiments, the multimer comprises two or more oligonucleotides linked end-to-end via an oligonucleotide-based linker (e.g., a poly-dT linker, a abasic linker). In some embodiments, the multimer comprises the 5' end of one oligonucleotide linked to the 3' end of another oligonucleotide. In some embodiments, the multimer comprises the 3' end of one oligonucleotide linked to the 3' end of another oligonucleotide. In some embodiments, the multimer comprises the 5' end of one oligonucleotide linked to the 5' end of another oligonucleotide. Additionally, in some embodiments, the multimer can include a branched structure comprising a plurality of oligonucleotides linked together by a branched linker.

[0261] Further examples of multimers that can be used in the complexes provided herein are disclosed, for example, in: US Patent Application No. 2015 / 0315588 A1, entitled Methods of delivering multiple targeting oligonucleotides to a cell using cleavable linkers, published November 5, 2015; US Patent Application No. 2015 / 0247141 A1, entitled Multimeric Oligonucleotide Compounds, published September 3, 2015; US Patent Application No. US 2011 / 0158937 A1, entitled Immunostimulatory Oligonucleotide Multimers, published June 30, 2011; and US Patent No. 5,693,773, entitled Triplex-Forming Antisense Oligonucleotides Having Abasic Linkers Targeting Nucleic Acids Comprising Mixed Sequences Of Purines And Pyrimidines, issued December 2, 1997. The entire contents of each of these are hereby incorporated by reference herein.

[0262] j. Gapmer In some embodiments, the oligonucleotides described herein are gapmers. Gapmer oligonucleotides generally have the formula 5'-X-Y-Z-3', having X and Z as flanking regions around the gap region Y. In some embodiments, the flanking region X of the formula 5'-X-Y-Z-3' is also referred to as the X region, flanking sequence X, 5' wing region X, or 5' wing fragment. In some embodiments, the flanking region Z of the formula 5'-X-Y-Z-3' is also referred to as the Z region, flanking sequence Z, 3' wing region Z, or 3' wing fragment. In some embodiments, the gap region Y of the formula 5'-X-Y-Z-3' is also referred to as the Y region, Y fragment, or gap fragment Y. In some embodiments, each nucleoside on the gap region Y is a 2'-deoxyribonucleoside, and neither the 5' wing region X nor the 3' wing region Z contains any 2'-deoxyribonucleoside. In some embodiments, the gapmer oligonucleotide comprises a region of complementarity to at least 15 consecutive nucleosides (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, or 20 consecutive nucleosides) of a target nucleic acid sequence provided herein (e.g., a transcript listed in Table 3 provided by any one of SEQ ID NOs: 392 to 702, or a target sequence of any of the oligonucleotides listed in Tables 5 to 19), and / or at least 15 consecutive nucleosides (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, or 20 consecutive nucleosides) of the nucleotide sequence of the oligonucleotide sequence in any one of Tables 5 to 19, wherein each thymine base (T) can be independently and optionally replaced by a uracil base (U), and each U can be independently and optionally replaced by a T.In some embodiments, the gapmer oligonucleotide comprises a region of complementarity to at least 15 consecutive nucleosides (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, or 20 consecutive nucleosides) of a target nucleic acid sequence provided herein (e.g., a transcript listed in Table 4 provided by any one of SEQ ID NOs: 705-803). In some embodiments, the gapmer oligonucleotide comprises a region of complementarity to at least 15 consecutive nucleosides (e.g., at least 15, at least 16, at least 17, at least 18, at least 19, or 20 consecutive nucleosides) of a target nucleic acid sequence provided herein (e.g., a transcript listed in Table 3 or Table 4 provided by any one of SEQ ID NOs: 143-148, 167-169, 810-875, and 1059-1068).

[0263] In some embodiments, the Y region is a stretch of nucleotides capable of recruiting an RNase, such as RNase H, e.g., a region of 6 or more DNA nucleotides. In some embodiments, the gapmer binds to the target nucleic acid, at which point the RNase is recruited and can then cleave the target nucleic acid. In some embodiments, the Y region is flanked on both the 5' and 3' by regions X and Z that include 1 to 6 high-affinity modified nucleosides, e.g., 1 to 6 high-affinity modified nucleosides. Examples of high-affinity modified nucleosides include, but are not limited to, 2'-modified nucleosides (e.g., 2'-MOE, 2'O-Me, 2'-F) or 2'-4' bicyclic nucleosides (e.g., LNA, cEt, ENA). In some embodiments, the flanking sequences X and Z can be 1 to 20 nucleotides, 1 to 8 nucleotides, or 1 to 5 nucleotides in length. The flanking sequences X and Z can be of similar length or dissimilar length. In some embodiments, the gap fragment Y can be a nucleotide sequence 5 to 20 nucleotides, 5 to 15 nucleotides, 5 to 12 nucleotides, or 6 to 10 nucleotides in length.

[0264] In some embodiments, the gap region of the gapmer oligonucleotide can contain, in addition to DNA nucleosides, modified nucleosides known to be tolerated for efficient RNase H activity, such as C4'-substituted nucleosides, acyclic nucleosides, and arabino-configured nucleosides. In some embodiments, the gap region includes one or more unmodified intervening nucleosides. In some embodiments, one or both of the flanking regions each independently include at least two, at least three, at least four, at least five, or more internucleotide linkages (e.g., phosphorothioate internucleotide linkages or other linkages) between one or more nucleotides. In some embodiments, the gap region and the two flanking regions each independently include at least two, at least three, at least four, at least five, or more internucleotide linkages (e.g., phosphorothioate internucleotide linkages or other linkages) between one or more nucleotides.

[0265] Gapmers can be produced using suitable methods. Representative U.S. patents, U.S. patent publications, and PCT publications that teach the preparation of gapmers include, but are not limited to: U.S. Patent Nos. 5,013,830; 5,149,797; 5,220,007; 5,256,775; 5,366,878; 5,403,711; 5,491,133; 5,565,350; 5,623,065; 5,652,355; 5,652,356; 5,700,922; 5,898,031; 7,015,315; 7,101,993; 7,399,845; 7,432,250; 7,569,686; 7,683,036; 7,750,131; 8,580,756; 9,045,754; 9,428,534; 9,695,418; 10,017,764; 10,260,069; 9,428,534; 8,580,756; U.S. Patent Publication Nos. US20050074801, US20090221685; US20090286969, US20100197762, and US20110112170; PCT Publication Nos. WO2004069991; WO2005023825; WO2008049085, and WO2009090182; and EP Patent No. EP2,149,605. Each of these is hereby incorporated by reference in its entirety.

[0266] In some embodiments, the gapmer is 10 to 40 nucleotides in length. For example, the gapmer can be 10 to 40, 10 to 35, 10 to 30, 10 to 25, 10 to 20, 10 to 15, 15 to 40, 15 to 35, 15 to 30, 15 to 25, 15 to 20, 20 to 40, 20 to 35, 20 to 30, 20 to 25, 25 to 40, 25 to 35, 25 to 30, 30 to 40, 30 to 35, or 35 to 40 nucleotides in length. In some embodiments, the gapmer is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 nucleotides in length.

[0267] In some embodiments, the gap region Y on the gapmer is 5 to 20 nucleotides in length. For example, the gap region Y can be 5 to 20, 5 to 15, 5 to 10, 10 to 20, 10 to 15, or 15 to 20 nucleotides in length. In some embodiments, the gap region Y is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length. In some embodiments, each nucleotide on the gap region Y is a 2'-deoxyribonucleotide. In some embodiments, all nucleotides on the gap region Y are 2'-deoxyribonucleotides. In some embodiments, one or more of the nucleotides on the gap region Y are modified nucleotides (e.g., 2'-modified nucleotides such as those described herein). In some embodiments, one or more cytosines on the gap region Y are optionally 5-methyl-cytosine. In some embodiments, each cytosine on the gap region Y is 5-methyl-cytosine.

[0268] In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) are each, independently, 1 to 20 nucleosides in length. For example, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) can each, independently, be 1 to 20, 1 to 15, 1 to 10, 1 to 7, 1 to 5, 1 to 3, 1 to 2, 2 to 5, 2 to 7, 3 to 5, 3 to 7, 5 to 20, 5 to 15, 5 to 10, 10 to 20, 10 to 15, or 15 to 20 nucleotides in length. In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) are each, independently, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleosides in length. In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) are of the same length. In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) are of different lengths. In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) is longer than the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format). In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) is shorter than the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format).

[0269] In some embodiments, the gapmer is 5-10-5, 4-12-4, 3-14-3, 2-16-2, 1-18-1, 3-10-3, 2-10-2, 1-10-1, 2-8-2, 4-6-4, 3-6-3, 2-6-2, 4-7-4, 3-7-3, 2-7-2, 4-8-4, 3-8-3, 2-8-2, 1-8-1, 2-9-2, 1-9-1, 2-10-2, 1-10-1, 1-12-1, 1-16-1, 2-15-1, 1-15-2, 1-14-3, 3-14-1, 2-14-2, 1-13-4, 4-13-1, 2-13-3, 3-13-2, 1-12-5, 5-12-1, 2-12-4, 4-12-2, 3-12-3, 1-11-6, 6-11-1, 2-11-5, 5-11-2, 3-11-4, 4-11-3, 1-17-1, 2-16-1, 1-16-2, 1-15-3, 3-15-1, 2-15-2, 1-14-4, 4-14-1, 2-14-3, 3-14-2, 1-13-5, 5-13-1, 2-13-4, 4-13-2, 3-13-3, 1-12-6, 6-12-1, 2-12-5, 5-12-2, 3-12-4, 4-12-3, 1-11-7, 7-11-1, 2-11-6, 6-11-2, 3-11-5, 5-11-3, 4-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 1-16-3, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 5-14-1, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 3-16-1, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4,1-19-1、1-18-2、2-18-1、1-17-3、3-17-1、2-17-2、1-16-4、4-16-1、2-16-3、3-16-2、1-15-5、2-15-4、4-15-2、3-15-3、1-14-6、6-14-1、2-14-5、5-14-2、3-14-4、4-14-3、1-13-7、7-13-1、2-13-6、6-13-2、3-13-5、5-13-3、4-13-4、1-12-8、8-12-1、2-12-7、7-12-2、3-12-6、6-12-3、4-12-5、5-12-4、2-11-8、8-11-2、3-11-7、7-11-3、4-11-6、6-11-4、5-11-5、1-20-1、1-19-2、2-19-1、1-18-3、3-18-1、2-18-2、1-17-4、4-17-1、2-17-3、3-17-2、1-16-5、2-16-4、4-16-2、3-16-3、1-15-6、6-15-1、2-15-5、5-15-2、3-15-4、4-15-3、1-14-7、7-14-1、2-14-6、6-14-2、3-14-5、5-14-3、4-14-4、1-13-8、8-13-1、2-13-7、7-13-2、3-13-6、6-13-3、4-13-5、5-13-4、2-12-8、8-12-2、3-12-7、7-12-3、4-12-6、6-12-4、5-12-5、3-11-8、8-11-3、4-11-7、7-11-4、5-11-6、6-11-5、1-21-1、1-20-2、2-20-1、1-20-3、3-19-1、2-19-2、1-18-4、4-18-1、2-18-3、3-18-2、1-17-5、2-17-4、4-17-2、3-17-3、1-16-6、6-16-1、2-16-5、5-16-2、3-16-4、4-16-3、1-15-7、7-15-1、2-15-6、6-15-2、3-15-5、5-15-3、4-15-4、1-14-8、8-14-1、2-14-7、7-14-2、3-14-6、6-14-3、4-14-5、5-14-4、2-13-8、8-13-2、3-13-7、7-13-3、4-13-6、6-13-4、5-13-5、1-12-10、10-12-1、2-12-9、9-12-2、3-12-8、8-12-3、4-12-7、7-12-4、5-12-6、6-12-5、Comprising 5'-X-Y-Z-3' of 4-11-8, 8-11-4, 5-11-7, 7-11-5, 6-11-6, 1-22-1, 1-21-2, 2-21-1, 1-21-3, 3-20-1, 2-20-2, 1-19-4, 4-19-1, 2-19-3, 3-19-2, 1-18-5, 2-18-4, 4-18-2, 3-18-3, 1-17-6, 6-17-1, 2-17-5, 5-17-2, 3-17-4, 4-17-3, 1-16-7, 7-16-1, 2-16-6, 6-16-2, 3-16-5, 5-16-3, 4-16-4, 1-15-8, 8-15-1, 2-15-7, 7-15-2, 3-15-6, 6-15-3, 4-15-5, 5-15-4, 2-14-8, 8-14-2, 3-14-7, 7-14-3, 4-14-6, 6-14-4, 5-14-5, 3-13-8, 8-13-3, 4-13-7, 7-13-4, 5-13-6, 6-13-5, 4-12-8, 8-12-4, 5-12-7, 7-12-5, 6-12-6, 5-11-8, 8-11-5, 6-11-7, or 7-11-6. The numbers indicate the number of nucleotides in the X, Y, and Z regions on the 5'-X-Y-Z-3' gapmer.,

[0270] In some embodiments, one or more nucleosides on the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) or the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) are modified nucleosides (e.g., high-affinity modified nucleosides). In some embodiments, the modified nucleoside (e.g., high-affinity modified nucleoside) is a 2'-modified nucleoside. In some embodiments, the 2'-modified nucleoside is a 2'-4' bicyclic nucleoside or a non-bicyclic 2'-modified nucleoside. In some embodiments, the high-affinity modified nucleoside is a 2'-4' bicyclic nucleoside (e.g., LNA, cEt, or ENA) or a non-bicyclic 2'-modified nucleoside (e.g., 2'-fluoro (2'-F), 2'-O-methyl (2'-O-Me), 2'-O-methoxyethyl (2'-MOE), 2'-O-aminopropyl (2'-O-AP), 2'-O-dimethylaminoethyl (2'-O-DMAOE), 2'-O-dimethylaminopropyl (2'-O-DMAP), 2'-O-dimethylaminoethyloxyethyl (2'-O-DMAEOE), or 2'-O-N-methylacetamide (2'-O-NMA)).

[0271] In some embodiments, one or more nucleosides on the 5' wing region (X in the 5'-X-Y-Z-3' format) of the gapmer are high-affinity modified nucleosides. In some embodiments, each nucleoside on the 5' wing region (X in the 5'-X-Y-Z-3' format) of the gapmer is a high-affinity modified nucleoside. In some embodiments, one or more nucleosides on the 3' wing region (Z in the 5'-X-Y-Z-3' format) of the gapmer are high-affinity modified nucleosides. In some embodiments, each nucleoside on the 3' wing region (Z in the 5'-X-Y-Z-3' format) of the gapmer is a high-affinity modified nucleoside. In some embodiments, one or more nucleosides on the 5' wing region (X in the 5'-X-Y-Z-3' format) of the gapmer are high-affinity modified nucleosides and one or more nucleosides on the 3' wing region (Z in the 5'-X-Y-Z-3' format) of the gapmer are high-affinity modified nucleosides. In some embodiments, each nucleoside on the 5' wing region (X in the 5'-X-Y-Z-3' format) of the gapmer is a high-affinity modified nucleoside and each nucleoside on the 3' wing region (Z in the 5'-X-Y-Z-3' format) of the gapmer is a high-affinity modified nucleoside.

[0272] In some embodiments, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) contains the same high-affinity nucleosides as the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format). For example, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) may contain one or more non-bicyclic 2'-modified nucleosides (e.g., 2'-MOE or 2'-O-Me). In another example, the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) may contain one or more 2'-4' bicyclic nucleosides (e.g., LNA or cEt). In some embodiments, each nucleoside on the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) is a non-bicyclic 2'-modified nucleoside (e.g., 2'-MOE or 2'-O-Me). In some embodiments, each nucleoside on the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) is a 2'-4' bicyclic nucleoside (e.g., LNA or cEt).

[0273] In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently 1 to 7 (e.g., 1, 2, 3, 4, 5, 6, or 7) nucleotides in length, Y is 6 to 10 (e.g., 6, 7, 8, 9, or 10) nucleotides in length, and where each nucleotide on X and Z is an acyclic 2’-modified nucleotide (e.g., 2’-MOE or 2’-O-Me), and each nucleotide on Y is a 2’-deoxyribonucleotide. In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently 1 to 7 (e.g., 1, 2, 3, 4, 5, 6, or 7) nucleotides in length, Y is 6 to 10 (e.g., 6, 7, 8, 9, or 10) nucleotides in length, and where each nucleotide on X and Z is a 2’-4’ bicyclic nucleotide (e.g., LNA or cEt), and each nucleotide on Y is a 2’-deoxyribonucleotide. In some embodiments, the 5’ wing region of the gapmer (X in the 5’-X-Y-Z-3’ formula) comprises different high-affinity nucleotides from the 3’ wing region of the gapmer (Z in the 5’-X-Y-Z-3’ formula). For example, the 5’ wing region of the gapmer (X in the 5’-X-Y-Z-3’ formula) may comprise one or more acyclic 2’-modified nucleotides (e.g., 2’-MOE or 2’-O-Me), and the 3’ wing region of the gapmer (Z in the 5’-X-Y-Z-3’ formula) may comprise one or more 2’-4’ bicyclic nucleotides (e.g., LNA or cEt). In another example, the 3’ wing region of the gapmer (Z in the 5’-X-Y-Z-3’ formula) may comprise one or more acyclic 2’-modified nucleotides (e.g., 2’-MOE or 2’-O-Me), and the 5’ wing region of the gapmer (X in the 5’-X-Y-Z-3’ formula) may comprise one or more 2’-4’ bicyclic nucleotides (e.g., LNA or cEt).

[0274] In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently nucleosides having a length of 1 to 7 (by way of example, 1, 2, 3, 4, 5, 6, or 7), Y is a nucleoside having a length of 6 to 10 (by way of example, 6, 7, 8, 9, or 10), where each nucleoside on X is an acyclic 2’-modified nucleoside (by way of example, 2’-MOE or 2’-O-Me), each nucleoside on Z is a 2’-4’ bicyclic nucleoside (by way of example, LNA or cEt), and each nucleoside on Y is a 2’-deoxyribonucleoside. In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently nucleosides having a length of 1 to 7 (by way of example, 1, 2, 3, 4, 5, 6, or 7), Y is a nucleoside having a length of 6 to 10 (by way of example, 6, 7, 8, 9, or 10), where each nucleoside on X is a 2’-4’ bicyclic nucleoside (by way of example, LNA or cEt), each nucleoside on Z is an acyclic 2’-modified nucleoside (by way of example, 2’-MOE or 2’-O-Me), and each nucleoside on Y is a 2’-deoxyribonucleoside.

[0275] In some embodiments, the 5’ wing region of the gapmer (X in the 5’-X-Y-Z-3’ formula) comprises one or more acyclic 2’-modified nucleosides (by way of example, 2’-MOE or 2’-O-Me) and one or more 2’-4’ bicyclic nucleosides (by way of example, LNA or cEt). In some embodiments, the 3’ wing region of the gapmer (Z in the 5’-X-Y-Z-3’ formula) comprises one or more acyclic 2’-modified nucleosides (by way of example, 2’-MOE or 2’-O-Me) and one or more 2’-4’ bicyclic nucleosides (by way of example, LNA or cEt). In some embodiments, both the 5’ wing region of the gapmer (X in the 5’-X-Y-Z-3’ formula) and the 3’ wing region of the gapmer (Z in the 5’-X-Y-Z-3’ formula) comprise one or more acyclic 2’-modified nucleosides (by way of example, 2’-MOE or 2’-O-Me) and one or more 2’-4’ bicyclic nucleosides (by way of example, LNA or cEt).

[0276] In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently 2 to 7 (e.g., 2, 3, 4, 5, 6, or 7) nucleotides in length, Y is 6 to 10 (e.g., 6, 7, 8, 9, or 10) nucleotides in length, where at least one (e.g., 1, 2, 3, 4, 5, or 6), but not all, of positions 1, 2, 3, 4, 5, 6, or 7 on X (where the most 5’ position is position 1) is a non-bicyclic 2’-modified nucleotide (e.g., 2’-MOE or 2’-O-Me), where the remainder of the nucleotides on both X and Z are 2’-4’ bicyclic nucleotides (e.g., LNA or cEt), and where each nucleotide on Y is a 2’-deoxyribonucleotide. In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently 2 to 7 (e.g., 2, 3, 4, 5, 6, or 7) nucleotides in length, Y is 6 to 10 (e.g., 6, 7, 8, 9, or 10) nucleotides in length, where at least one (e.g., 1, 2, 3, 4, 5, or 6), but not all, of positions 1, 2, 3, 4, 5, 6, or 7 on Z (where the most 5’ position is position 1) is a non-bicyclic 2’-modified nucleotide (e.g., 2’-MOE or 2’-O-Me), where the remainder of the nucleotides on both X and Z are 2’-4’ bicyclic nucleotides (e.g., LNA or cEt), and where each nucleotide on Y is a 2’-deoxyribonucleotide.In some embodiments, the gapmer comprises a 5’-X-Y-Z-3’ configuration, where X and Z are independently nucleosides of length 2 to 7 (by way of example, 2, 3, 4, 5, 6, or 7), Y is a nucleoside of length 6 to 10 (by way of example, 6, 7, 8, 9, or 10), where at least one (by way of example, 1, 2, 3, 4, 5, or 6) but not all of positions 1, 2, 3, 4, 5, 6, or 7 (the most 5’ position being position 1) on X and at least one (by way of example, 1, 2, 3, 4, 5, or 6) but not all of positions 1, 2, 3, 4, 5, 6, or 7 (the most 5’ position being position 1) on Z are acyclic 2’-modified nucleosides (by way of example, 2’-MOE or 2’-O-Me), where the remainder of the nucleosides on both X and Z are 2’-4’ bicyclic nucleosides (by way of example, LNA or cEt), where each nucleoside on Y is a 2’-deoxyribonucleoside.

[0277] Non-limiting examples of gapmer constructs having a mix of acyclic 2'-modified nucleosides (e.g., 2'-MOE or 2'-O-Me) and 2'-4' bicyclic nucleosides (e.g., LNA or cEt) in the 5' wing region of the gapmer (X in the 5'-X-Y-Z-3' format) and / or the 3' wing region of the gapmer (Z in the 5'-X-Y-Z-3' format) include: BBB-(D)n-BBBAA; KKK-(D)n-KKKAA; LLL-(D)n-LLLAA; BBB-(D)n-BBBEE; KKK-(D)n-KKKEE; LLL-(D)n-LLLEE; BBB-(D)n-BBBAA; KKK-(D)n-KKKAA; LLL-(D)n-LLLAA; BBB-(D)n-BBBEE; KKK-(D)n-KKKEE; LLL-(D)n-LLLEE; BBB-(D)n-BBBAAA; KKK-(D)n-KKKAAA; LLL-(D)n-LLLAAA; BBB-(D)n-BBBEEE; KKK-(D)n-KKKEEE; LLL-(D)n-LLLEEE; BBB-(D)n-BBBAAA; KKK-(D)n-KKKAAA; LLL-(D)n-LLLAAA; BBB-(D)n-BBBEEE; KKK-(D)n-KKKEEE; LLL-(D)n-LLLEEE; BABA-(D)n-ABAB; KAKA-(D)n-AKAK; LALA-(D)n-ALAL; BEBE-(D)n-EBEB; KEKE-(D)n-EKEK; LELE-(D)n-ELEL; BABA-(D)n-ABAB; KAKA-(D)n-AKAK; LALA-(D)n-ALAL; BEBE-(D)n-EBEB; KEKE-(D)n-EKEK; LELE-(D)n-ELEL; ABAB-(D)n-ABAB; AKAK-(D)n-AKAK; ALAL-(D)n-ALAL; EBEB-(D)n-EBEB; EKEK-(D)n-EKEK; ELEL-(D)n-ELEL; ABAB-(D)n-ABAB; AKAK-(D)n-AKAK; ALAL-(D)n-ALAL; EBEB-(D)n-EBEB; EKEK-(D)n-EKEK; ELEL-(D)n-ELEL; AABB-(D)n-BBAA; BBAA-(D)n-AABB; AAKK-(D)n-KKAA; AALL-(D)n-LLAA; EEBB-(D)n-BBEE;EEKK-(D)n-KKEE;EELL-(D)n-LLEE;AABB-(D)n-BBAA;AAKK-(D)n-KKAA;AALL-(D)n-LLAA;EEBB-(D)n-BBEE;EEKK-(D)n-KKEE;EELL-(D)n-LLEE;BBB-(D)n-BBA;KKK-(D)n-KKA;LLL-(D)n-LLA;BBB-(D)n-BBE;KKK-(D)n-KKE;LLL-(D)n-LLE;BBB-(D)n-BBA;KKK-(D)n-KKA;LLL-(D)n-LLA;BBB-(D)n-BBE;KKK-(D)n-KKE;LLL-(D)n-LLE;BBB-(D)n-BBA;KKK-(D)n-KKA;LLL-(D)n-LLA;BBB-(D)n-BBE;KKK-(D)n-KKE;LLL-(D)n-LLE;ABBB-(D)n-BBBA;AKKK-(D)n-KKKA;ALLL-(D)n-LLLA;EBBB-(D)n-BBBE;EKKK-(D)n-KKKE;ELLL-(D)n-LLLE;ABBB-(D)n-BBBA;AKKK-(D)n-KKKA;ALLL-(D)n-LLLA;EBBB-(D)n-BBBE;EKKK-(D)n-KKKE;ELLL-(D)n-LLLE;ABBB-(D)n-BBBAA;AKKK-(D)n-KKKAA;ALLL-(D)n-LLLAA;EBBB-(D)n-BBBEE;EKKK-(D)n-KKKEE;ELLL-(D)n-LLLEE;ABBB-(D)n-BBBAA;AKKK-(D)n-KKKAA;ALLL-(D)n-LLLAA;EBBB-(D)n-BBBEE;EKKK-(D)n-KKKEE;ELLL-(D)n-LLLEE;AABBB-(D)n-BBB;AAKKK-(D)n-KKK;AALLL-(D)n-LLL;EEBBB-(D)n-BBB;EEKKK-(D)n-KKK;EELLL-(D)n-LLL;AABBB-(D)n-BBB;AAKKK-(D)n-KKK;AALLL-(D)n-LLL;EEBBB-(D)n-BBB;EEKKK-(D)n-KKK;EELLL-(D)n-LLL;AABBB-(D)n-BBBA;AAKKK-(D)n-KKKA;AALLL-(D)n-LLLA;EEBBB-(D)n-BBBE;EEKKK-(D)n-KKKE;EELLL-(D)n-LLLE; AABBB-(D)n-BBBA; AAKKK-(D)n-KKKA; AALLL-(D)n-LLLA; EEBBB-(D)n-BBBE; EEKKK-(D)n-KKKE; EELLL-(D)n-LLLE; ABBAABB-(D)n-BB; AKKAAKK-(D)n-KK; ALLAALLL-(D)n-LL; EBBEEBB-(D)n-BB; EKKEEKK-(D)n-KK; ELLEELL-(D)n-LL; ABBAABB-(D)n-BB; AKKAAKK-(D)n-KK; ALLAALL-(D)n-LL; EBBEEBB-(D)n-BB; EKKEEKK-(D)n-KK; ELLEELL-(D)n-LL; ABBABB-(D)n-BBB; AKKAKK-(D)n-KKK; ALLALLL-(D)n-LLL; EBBEBB-(D)n-BBB; EKKEKK-(D)n-KKK; ELLELL-(D)n-LLL; ABBABB-(D)n-BBB; AKKAKK-(D)n-KKK; ALLALL-(D)n-LLL; EBBEBB-(D)n-BBB; EKKEKK-(D)n-KKK; ELLELL-(D)n-LLL; EEEK-(D)n-EEEEEEEE; EEK-(D)n-EEEEEEEEE; EK-(D)n-EEEEEEEEEE; EK-(D)n-EEEKK; K-(D)n-EEEKEKE; K-(D)n-EEEKEKEE; K-(D)n-EEKEK; EK-(D)n-EEEEKEKE; EK-(D)n-EEEKEK; EEK-(D)n-KEEKE; EK-(D)n-EEKEK; EK-(D)n-KEEK; EEK-(D)n-EEEKEK; EK-(D)n-KEEEKEE; EK-(D)n-EEKEKE; EK-(D)n-EEEKEKE; and EK-(D)n-EEEEKEK; wherein, "A" represents a 2'-modified nucleoside; "B" represents a 2',4'-bicyclic nucleoside; "K" represents a constrained ethyl nucleoside (cEt); "L" represents an LNA nucleoside; "E" represents a 2'-MOE-modified ribonucleoside; "D" represents a 2'-deoxyribonucleoside; "n" represents the length of the gap fragment (Y in the 5'-X-Y-Z-3' configuration), which is an integer from 1 to 20.;

[0278] In some embodiments, any one of the gapmers described herein includes one or more modified nucleoside linkages (e.g., phosphorothioate linkages) in each of the X, Y, and Z regions. In some embodiments, each internucleoside linkage in any one of the gapmers described herein is a phosphorothioate linkage. In some embodiments, each of the X, Y, and Z regions independently includes a mix of phosphorothioate linkages and phosphodiester linkages. In some embodiments, each internucleoside linkage in the gap region Y is a phosphorothioate linkage, the 5' wing region X includes a mix of phosphorothioate linkages and phosphodiester linkages, and the 3' wing region Z includes a mix of phosphorothioate linkages and phosphodiester linkages.

[0279] Polypeptide payload In some embodiments, polypeptides (e.g., peptides, proteins including but not limited to enzymes, antibodies, etc.) are useful for the treatment of various CNS diseases and disorders. For example, polypeptides may be useful for modulating the expression or activity of various genes involved in CNS diseases and disorders, such as by modulating the expression or activity of proteins involved in a CNS disease or disorder. In one non-limiting example, an enzyme that modifies, degrades, or otherwise affects a particular biological molecule (e.g., a protein or nucleic acid) may be useful for the treatment of a CNS disease or disorder in which that particular biological molecule is involved. Polypeptides may be used to treat various CNS diseases and disorders, such as by facilitating delivery of the polypeptide into cells of the CNS. In some embodiments, the polypeptides disclosed herein may be delivered into cells of the CNS using the complexes disclosed herein (e.g., an anti-TfR1 antibody complex comprising a polypeptide).

[0280] In some embodiments, the polypeptide is useful for the treatment of neuromuscular diseases or disorders (by way of example, Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy); amyotrophic lateral sclerosis; Parkinson's disease; Huntington's disease; Alzheimer's disease; epilepsy; pain disorders; glycogen synthesis disorders; neurodegeneration; small fiber neuropathy; phenotypes related to nociception; Alexander disease; Angelman syndrome; autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis.

[0281] In some embodiments, the polypeptide is useful for the treatment of essential tremor and / or hereditary dystonia.

[0282] In some embodiments, the polypeptide is useful for the treatment of spinocerebellar ataxia, motor neuron diseases, Draver syndrome, Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, Gaucher disease types II and III, and / or Rett syndrome.

[0283] In some embodiments, the polypeptide is useful for regulating one or more genes associated with a CNS disease or disorder. In some embodiments, one or more genes associated with a CNS disease or disorder are DMPK, DMD, SMN, FXN, SOD1, C9orf72, ATXN2, FUS, LRRK2, SNCA, HTT, MSH3, TREM2, APOE, MAPT, APP, GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, SCN9A, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, or PCDH19. In some embodiments, one or more genes associated with a CNS disease or disorder are TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1. In some embodiments, one or more genes associated with a CNS disease or disorder are PIKFYVE, SYF2, UNC13A, ATXN1, ATXN3, GRN, GRIN2A, TPP1, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2.

[0284] Small molecule payload In some embodiments, the small molecule is useful for treating various CNS diseases and disorders. For example, the small molecule can be useful for regulating the expression or activity of various genes involved in CNS diseases and disorders, such as by regulating the expression or activity of a protein involved in a CNS disease or disorder. In one non-limiting example, a small molecule that increases, decreases, or otherwise affects the expression of a particular biological molecule (e.g., a protein or nucleic acid) can be useful for treating a CNS disease or disorder in which that particular biological molecule is involved. The small molecule can be used to treat various CNS diseases and disorders, such as by facilitating delivery of the small molecule into cells of the CNS. In some embodiments, the small molecules disclosed herein can be delivered into cells of the CNS using the complexes disclosed herein (e.g., an anti-TfR1 antibody complex comprising a small molecule).

[0285] In some embodiments, the small molecule is useful for the treatment of a neuromuscular disease or disorder (by way of example, Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy); amyotrophic lateral sclerosis; Parkinson's disease; Huntington's disease; Alzheimer's disease; epilepsy; a pain disorder; a glycogen synthesis disorder; neurodegeneration; small fiber neuropathy; a phenotype related to nociception; Alexander disease; Angelman syndrome; an autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis.

[0286] In some embodiments, the small molecule is useful for the treatment of essential tremor and / or hereditary dystonia.

[0287] In some embodiments, the small molecule is useful for the treatment of spinocerebellar ataxia, a motor neuron disease, Dravet syndrome, Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, Gaucher disease types II and III, and / or Rett syndrome.

[0288] In some embodiments, the small molecule is useful for the regulation of one or more genes associated with a CNS disease or disorder. In some embodiments, one or more genes associated with a CNS disease or disorder are DMPK, DMD, SMN, FXN, SOD1, C9orf72, ATXN2, FUS, LRRK2, SNCA, HTT, MSH3, TREM2, APOE, MAPT, APP, GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, SCN9A, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, or PCDH19. In some embodiments, one or more genes associated with a CNS disease or disorder are TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1. In some embodiments, one or more genes associated with a CNS disease or disorder are PIKFYVE, SYF2, UNC13A, ATXN1, ATXN3, GRN, GRIN2A, TPP1, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2.

[0289] The compounds described herein (e.g., small molecule payloads) can contain one or more asymmetric centers and, therefore, can exist in various stereoisomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein can be in the form of individual enantiomers, diastereomers, or geometric isomers, or in the form of a mixture of stereoisomers. This includes racemic mixtures and mixtures enriched in one or more stereoisomers. Isomers can be isolated from the mixtures by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or the preferred isomers can be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, E.L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, S.H., Tables of Resolving Agents and Optical Resolutions p. 268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The disclosure also includes the compounds as individual isomers substantially free of other isomers and alternatively as mixtures of various isomers.

[0290] Gene therapy payload In some embodiments, gene therapy payloads (e.g., nucleic acids encoding bioactive or other therapeutic molecules) are useful for treating various CNS diseases and disorders. For example, gene therapy can be useful for modulating the expression or activity of various genes involved in CNS diseases and disorders, such as by encoding a protein involved in a CNS disease or disorder. In one non-limiting example, a gene therapy payload encoding a particular biological molecule (e.g., a protein or nucleic acid) can be useful for treating a CNS disease or disorder in which that particular biological molecule is involved (e.g., a disease or disorder in which abnormally low expression of the biological molecule or expression of an inactive form of the biological molecule is involved in its etiology). Gene therapy can be used to treat various CNS diseases and disorders, for example, by facilitating delivery of the gene therapy payload into cells of the CNS. In some embodiments, the gene therapy disclosed herein can be delivered into cells of the CNS using the complexes disclosed herein (e.g., an anti-TfR1 antibody complex comprising a gene therapy payload).

[0291] In some embodiments, gene therapy is useful for treating neuromuscular diseases or disorders (e.g., Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy); amyotrophic lateral sclerosis; Parkinson's disease; Huntington's disease; Alzheimer's disease; epilepsy; pain disorders; glycogen synthesis disorders; neurodegeneration; small fiber neuropathy; phenotypes related to nociception; Alexander disease; Angelman syndrome; autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis.

[0292] In some embodiments, gene therapy is useful for treating essential tremor and / or hereditary dystonia.

[0293] In some embodiments, gene therapy is useful for the treatment of spinocerebellar ataxia, motor neuron disease, Dravet syndrome, Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, Gaucher disease types II and III, and / or Rett syndrome.

[0294] In some embodiments, gene therapy is useful for the regulation of one or more genes associated with a CNS disease or disorder. In some embodiments, one or more genes associated with a CNS disease or disorder are DMPK, DMD, SMN, FXN, SOD1, C9orf72, ATXN2, FUS, LRRK2, SNCA, HTT, MSH3, TREM2, APOE, MAPT, APP, GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, SCN9A, SCN1A, SCN2A, SCN8A, CLN3, GRIA1, or PCDH19. In some embodiments, one or more genes associated with a CNS disease or disorder are TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1. In some embodiments, one or more genes associated with a CNS disease or disorder are PIKFYVE, SYF2, UNC13A, ATXN1, ATXN3, GRN, GRIN2A, TPP1, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2.

[0295] Molecular payload for the treatment of ALS Various molecular payloads, including oligonucleotides, polypeptides (such as peptides, proteins, enzymes, antibodies, etc.), small molecules (such as small molecule inhibitors, etc.), and gene therapies (such as nucleic acids and / or nucleic acid vectors encoding therapeutic molecules such as therapeutic proteins), may be useful for the treatment of ALS. In some embodiments, the molecular payload useful for the treatment of ALS may include a molecular payload that regulates (such as increases or decreases) the expression or activity of SOD1, ATXN2, C9orf72, and / or FUS.

[0296] Examples of oligonucleotides useful for the treatment of ALS, such as oligonucleotides that target (such as directly or indirectly regulate the expression or activity of) genes associated with ALS (such as SOD1, ATXN2, C9orf72, FUS, etc.), include those listed in Table 5 below. Each oligonucleotide provided in Table 5 may have any of the modification patterns disclosed herein.

Table 5-1

Table 5-2

[0297] Examples of oligonucleotides useful for the treatment of ALS, such as oligonucleotides that target (such as directly or indirectly regulate the expression or activity of) genes associated with ALS (such as PIKFYVE, SYF2, UNC13A, etc.), include those listed in Table 6 below. Each oligonucleotide provided in Table 6 may have any of the modification patterns disclosed herein.

Table 6-1

Table 6-2

Table 6-3

[0298] Examples of small molecules useful for the treatment of ALS include the following: [Chemical formula] And their pharmaceutically acceptable salts, co-crystals, tautomers, stereoisomers, solvates, hydrates, polymorphs, isotope-enriched compounds, and prodrugs. Additional examples of small molecules useful for the treatment of ALS include the following: [Chemical formula] Apilimod, APY0201, YM-201636, and their pharmaceutically acceptable salts, co-crystals, tautomers, stereoisomers, solvates, hydrates, polymorphs, isotope-enriched compounds, and prodrugs.

[0299] An example of a polypeptide useful for the treatment of ALS is (R)-2-amino-N-((S)-1-(((S)-5-amino-1-(3-benzyl-1,2,4-oxadiazol-5-yl)pentyl)amino)-3-(4-hydroxy-2,6-dimethylphenyl)-1-oxopropan-2-yl)-5-guanidinopentanamide.

[0300] Molecular payload targeting SOD1 The superoxide dismutase 1 (SOD1) gene and mutations therein are involved in ALS, which affects predominantly upper and lower motor neurons. Thus, in some embodiments, modulation of SOD1 expression and activity (e.g., by suppressing the expression and / or activity of mutant SOD1 protein) may have a therapeutic effect in subjects having ALS.

[0301] Oligonucleotide In some embodiments, SOD1 expression and / or activity can be modulated by use of oligonucleotides that target the SOD1 sequence.

[0302] In some embodiments, oligonucleotides useful for treating ALS, such as those that target SOD1 (e.g., that directly or indirectly modulate its expression or activity), include regions of complementarity to the SOD1 transcript provided in Table 3, provided by SEQ ID NO: 392, for example.

[0303] In some embodiments, examples of oligonucleotides useful for the treatment of ALS that target SOD1 (e.g., that directly or indirectly modulate its expression or activity) are provided below: Smith, et al., “Antisense oligonucleotide therapy for neurodegenerative disease” J. Clin. Invest. (2006) 116(8): 2290-96 doi:10.1172 / JCI25424; van Zundert, et al., “Silencing strategies for therapy of SOD1-mediated ALS” Neurosis. Lett. (2017) 636:32-39 doi:10.1016 / j.neulet.2016.07.059; US Patent Application Publication No. 20040091919A1, published May 13, 2004, entitled “Antisense Modulation of Superoxide Dismutase 1, Soluble Expression”; US Patent Application Publication No. 20090306005A1, published December 10, 2009, entitled “Compounds and methods for modulating expression of PCSK9”; US Patent Application Publication No. 20140378533A1, published December 25, 2014, entitled “Modulation of RNA by repeat targeting”; US Patent Application Publication No. 20150184154A1, published July 2, 2015, entitled “New Treatment for Neurodegenerative Diseases”; US Patent Application Publication No. 20160272976A1, published September 22, 2016, entitled “Products and Methods for Treatment of Familial Amyotrophic Lateral Sclerosis”; US Patent Application Publication No., published August 4, 2016, entitled “Compositions and Methods for Treating Amyotrophic Lateral Sclerosis”US Patent Application Publication No. 20160222391A1; US Patent Application Publication No. 20170037399A1, published on February 9, 2017, entitled "Chiral Design"; US Patent Application Publication No. 20170037410A1, published on February 9, 2017, entitled "Compositions for Modulating SOD-1 Expression"; US Patent Application Publication No. 20170152517A1, published on June 1, 2017, entitled "Treatment of Amyotrophic Lateral Sclerosis"; US Patent Application Publication No. 20160089453A1, published on March 31, 2016, entitled "RNA-Modulating Agents"; US Patent Application Publication No. 20180282732A1, published on October 4, 2018, entitled "Compositions and Methods of Treating Amyotrophic Lateral Sclerosis (ALS)"; International Patent Application Publication No. WO2016180784A1, published on November 17, 2016, entitled "Improved Treatments Using Oligonucleotides"; US Patent Application Publication No. 20180161357A1, published on June 14, 2018, entitled "MIR-155 Inhibitors for Treating Amyotrophic Lateral Sclerosis (ALS)"; US Patent Application Publication No. 20180195072A1, published on July 12, 2018, entitled "Nucleic acid molecules targeting superoxide dismutase 1 (sod1)"; US Patent Application Publication No. 20180216107A1, published on August 2, 2018, entitled "Oligonucleotide compositions and methods thereof"; US Patent Application Publication No., published on July 29, 2021, entitled "Oligonucleotide compositions and methods thereof"US Patent Application Publication No. 20210228615A1; US Patent Application Publication No. 20190167815A1, published on June 6, 2019, entitled "Methods and compositions for the treatment of rare diseases"; US Patent Application Publication No. 20210054383A1, published on February 25, 2021, entitled "Oligonucleotides for modulating tmem106b expression"; US Patent Application Publication No. 20210269881A1, published on September 2, 2021, entitled "Long non-coding RNAs (lncRNAs) for the diagnosis and therapeutics of brain disorders, in particular cognitive disorders"; US Patent Publication No. 10808247B2, published on October 20, 2020, entitled "Methods for treating neurological disorders using a synergistic small molecule and nucleic acids therapeutic approach"; US Patent Publication No. 11118179B2, published on September 14, 2021, entitled "Mixed tricyclo-DNA, 2’-modified RNA oligonucleotide compositions and uses thereof"; International Patent Application Publication No. WO2020198270A1, published on October 1, 2020, entitled "Compositions and methods for treating neurodegenerative disorders"; US Patent Application Publication No. 20220170025A1, published on June 2, 2022, entitled "Compositions and methods for inhibiting gene expression in the central nervous system"; US Patent Publication No., published on January 8, 2019, entitled "Compositions and methods for treating amyotrophic lateral sclerosis"10174328B2; International Patent Application Publication No. WO2020222182A1, published on November 5, 2020, entitled "Treatment for SOD1 associated disease"; International Patent Application Publication No. WO2020247419A2, published on December 10, 2020, entitled "Oligonucleotides and methods of use for treating neurological diseases"; International Patent Application Publication No. WO2021029896A1, published on February 18, 2021, entitled "Splice modulating oligonucleotides targeting receptor for advanced glycation end products and methods of use"; US Patent Application Publication No. 20220090036A1, published on March 24, 2022, entitled "Compositions and methods for the targeting of SOD1"; International Patent Application Publication No. WO2021108602A1, published on November 25, 2020, entitled "Methods and compositions for neuroprotection"; International Patent Application Publication No. WO2021156832A1, published on February 6, 2021, entitled "Use of miRNA-485 inhibitors for treating amyotrophic lateral sclerosis (ALS)"; US Patent Application Publication No. 20220073930A1, published on March 10, 2022, entitled "Compositions and methods for treating and preventing amyotrophic lateral sclerosis"; The entire contents of each of these are hereby incorporated by reference into this specification.

[0304] Certain oligonucleotides provided in this section may be useful in treating ALS by modulating the activity of genes and / or gene products other than the SOD1 gene / gene product, such as the C9orf72, ATXN2, and / or FUS gene / gene product.

[0305] Polypeptide In some embodiments, SOD1 expression and / or activity can be modulated by the use of polypeptides that can interact with SOD1 (e.g., to modulate its enzymatic activity) or SOD1 polypeptides.

[0306] In some embodiments, examples of polypeptides useful for the treatment of ALS (e.g., peptides; proteins such as enzymes; antibodies, etc.) are provided below: Martin, et al. “GNX-4728, a novel small molecule drug inhibitor of mitochondrial permeability transition, is therapeutic in a mouse model of amyotrophic lateral sclerosis” Front. Cell Neurosci. 8: article 433 (2014); doi: 10.3389 / fncel.2014.00433; U.S. Patent Application Publication No. 20090124993A1, published May 14, 2009, entitled “Treating neurological disorders”; U.S. Patent Application Publication No. 20140044722A1, published February 13, 2014, entitled “Anti-SOD1 Antibodies and Uses Thereof”; U.S. Patent Application Publication No. 20140301945, published October 9, 2014, entitled “Human Anti-SOD1 Antibodies”; International Patent Application Publication No. WO2013106672A1, published July 18, 2013, entitled “Methods and Compositions for the Treatment of Neurodegenerative Disease”; U.S. Patent Application Publication No. 20150184154A1, published July 2, 2015, entitled “New Treatment for Neurodegenerative Diseases”; U.S. Patent Application Publication No. 20150259391A1, published September 17, 2015, entitled “Treatment of Amyotrophic Lateral Sclerosis”; U.S. Patent Application Publication No., published April 28, 2016, entitled “Single Domain Antibodies Against SOD1 and Their Use in Medicine”US Patent Application Publication No. 20160115245A1; US Patent Application Publication No. 20190022179A1, published on January 24, 2019, entitled "Composition and method for treating amyotrophic lateral sclerosis"; US Patent Application Publication No. 20200247854A1, published on August 6, 2020, entitled "Pharmaceutical composition for preventing or treating neurodegenerative disease comprising nckap1 protein or gene encoding same"; US Patent Application Publication No. 20210206876A1, published on July 8, 2021, entitled "DPP3 binder directed to and binding to specific DPP3 - epitopes and its use in the prevention or treatment of diseases / acute conditions that are associated with oxidative stress"; International Patent Application Publication No. WO2019104311A1, published on May 31, 2019, entitled "Compositions and methods for suppressing neurological disease"; US Patent Application Publication No. 20210100869A1, published on April 8, 2021, entitled "Compositions and methods of using same for treating amyotrophic lateral sclerosis (ALS)"; US Patent Application Publication No. 20200172590A1, published on June 4, 2020, entitled "Methods of treating neurological diseases"; US Patent Application Publication No., published on June 3, 2021, entitled "Regenerating functional neurons for treatment of spinal cord injury and ALS"20210162002A1; US Patent Publication No. 10808247B2, published on October 20, 2020, entitled "Methods for treating neurological disorders using a synergistic small molecule and nucleic acids therapeutic approach"; US Patent Application Publication No. 20210284702A1, published on September 16, 2021, entitled "Fusion proteins comprising progranulin"; US Patent Application Publication No. 20220017634A1, published on January 20, 2022, entitled "Engineered bispecific proteins"; US Patent Application Publication No. 20220034907A1, published on February 3, 2022, entitled "Neurofilament protein for guiding therapeutic intervention in amyotrophic lateral sclerosis"; the entire contents of each of these are incorporated herein by reference.

[0307] Certain polypeptides provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the SOD1 gene / gene product, such as the C9orf72, ATXN2, and / or FUS gene / gene product.

[0308] Small molecule In some embodiments, SOD1 expression and / or activity can be modulated by the use of small molecules that can modulate SOD1 (e.g., to modulate its enzymatic activity or its expression).

[0309] In some embodiments, examples of small molecules useful for the treatment of ALS are provided below: U.S. Patent Application Publication No. 20040219552A1, published November 4, 2004, entitled "Novel Molecular Target for Neurotoxicity"; U.S. Patent Application Publication No. 20030130357A1, published July 10, 2003, entitled "Use of Polyamine Analogs for Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No. 20100152125A1, published June 17, 2010, entitled "Compositions and Methods for the Diagnosis, Treatment, and Prevention of Amyotrophic Lateral Sclerosis and Related Neurological Diseases"; U.S. Patent Application Publication No. 20100331417A1, published December 30, 2010, entitled "Treatment of Neural Diseases or Conditions"; U.S. Patent Application Publication No. 20110076236A1, published March 31, 2011, entitled "Compositions and Methods of Treatment Using Modulators of Motoneuron Diseases"; U.S. Patent Application Publication No. 20110166115A1, published July 7, 2011, entitled "Use of Mifepristone for the Treatment of Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No., published June 18, 2015, entitled "Compounds, Compositions and Methods for Treating or Preventing Neurodegenerative Disorders"US Patent Application Publication No. 20150164901A1, published on March 24, 2016, titled "Methods, Compositions and Kits for Promoting Motor Neuron Survival and Treating and Diagnosing Neurodegenerative Disorders"; US Patent Application Publication No. 20160082015A1, published on March 24, 2016, titled "Novel Methods for Treating Neurodegenerative Diseases"; US Patent Application Publication No. 20150210679A1, published on July 30, 2015, titled "Small Molecule Inhibitors of Superoxide Dismutase Expression"; International PCT Application Publication No. WO2021174167A1, published on September 2, 2021, titled "Compounds and methods for modulating splicing"; US Patent Application Publication No. 20180028520A1, published on February 1, 2018, titled "Methods and Pharmaceutical Compositions for Treatment of Amyotrophic Lateral Sclerosis"; International Patent Application Publication No. WO2016114655A1, published on July 21, 2016, titled "Treating neuromuscular or neurologic disease through reducing gabaergic and / or glycinergic inhibitory neurotransmitter overstimulation"; US Patent Application Publication No., published on June 8, 2017, titled "Methods of Using GM604 in Modulating ALS Disease Biomarkers Leading to Prognosis and Therapeutic Treatment for ALS Disease"US Patent Application Publication No. 20170157197A1, published on December 14, 2017, entitled "Diterpenoid derivatives and methods of use thereof"; US Patent Application Publication No. 20180289655A1, published on October 11, 2018, entitled "Methods and Compositions for the Intravenous Administration of Fumarates for the Treatment of Neurological Diseases"; US Patent Application Publication No. 20170226127A1, published on August 10, 2017, entitled "Compound, compositions, and methods"; US Patent Application Publication No. 20170362206A1, published on June 15, 2017, entitled "Compound, compositions, and methods"; US Patent Application Publication No. 20180327391A1, published on November 15, 2018, entitled "Compound, compositions, and methods"; US Patent Application Publication No. 20190300537A1, published on October 3, 2019, entitled "Compound, compositions, and methods"; US Patent Application Publication No. 20190194170A1, published on June 27, 2019, entitled "Polymorphs and solid forms of a pyrimidinylamino-pyrazole compound, and methods of production"; US Patent Publication No. 9669014B2, published on June 6, 2017, entitled "Small molecule inhibitors of superoxide dismutase expression"; US Patent Application Publication No., published on May 6, 2021, entitled "Modulators of eukaryotic initiation factor 2"20210130308A1; US Patent Application Publication No. 20180353480A1, published on December 13, 2018, entitled "Isoxazolidine derived inhibitors of receptor interacting protein kinase 1 (RIPK1)"; US Patent Application Publication No. 20200079784A1, published on March 12, 2020, entitled "Compound, compositions, and methods"; US Patent Application Publication No. 20200331900A1, published on October 22, 2020, entitled "Compounds, compositions, and methods"; US Patent Application Publication No. 20210147435A1, published on May 20, 2021, entitled "Compounds, compositions, and methods"; US Patent Application Publication No. 20210292311A1, published on September 23, 2021, entitled "Compounds, compositions, and methods"; US Patent Application Publication No. 20220177456A1, published on June 9, 2022, entitled "Compounds, compositions, and methods"; US Patent Application Publication No. 20200087319A1, published on March 19, 2020, entitled "Kinase Inhibitors and Uses Thereof"; US Patent Application Publication No. 20190359634A1, published on November 28, 2019, entitled "ASK1 inhibiting agents"; US Patent Application Publication No. 20200368267A1, published on November 26, 2020, entitled "Prophylactic and / or therapeutic agent for amyotrophic lateral sclerosis"; US Patent Application Publication No. 20210115020A1, published on April 22, 2021, entitled "ASK1 inhibiting agents"; US Patent Application Publication No. 20210115020A1, published on October 14, 2021, entitled "ASK1 inhibiting agents"20210317103A1; US Patent Application Publication No. 20210300946A1, published on September 30, 2021, entitled "Pyridine Macrocycle Compounds as ASK1 Inhibiting Agents"; US Patent Application Publication No. 20210353611A1, published on November 18, 2021, entitled "Methods of treating amyotrophic lateral sclerosis"; US Patent Application Publication No. US20210023062A1, published on January 28, 2021, entitled "Compositions and Methods for the Treatment of Amyotrophic Lateral Sclerosis, Parkinson’s Disease, Parkinson’s Disease with Dementia, Dementia with Lewy Bodies, and Multiple System Atrophy"; International Patent Application Publication No. WO2021040627A1, published on March 4, 2021, entitled "A method of promoting survival and / or function of a motor neuron and related agents, uses and methods"; the entire contents of each of these are incorporated herein by reference.

[0310] In some embodiments, the small molecule is a pharmaceutically acceptable salt, co-crystal, tautomer, stereoisomer, solvate, hydrate, polymorph, isotopically enriched compound, or prodrug of the small molecules provided herein.

[0311] Certain small molecules provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the SOD1 gene / gene product, such as the C9orf72, ATXN2, and / or FUS gene / gene product.

[0312] Gene Therapy In some embodiments, SOD1 expression and / or activity can be modulated by the use of gene therapy that can regulate SOD1 (by way of example, by delivery of nucleic acids encoding SOD1, or other molecules that interact with SOD1).

[0313] In some embodiments, provided below are gene therapies useful for the treatment of ALS, for example, those involving the administration of compounds encoding useful therapeutic agents: U.S. Patent Application Publication No. 20030161814A1, published August 28, 2003, entitled "Adeno-Associated Virus-Mediated Delivery of GDNF to Skeletal Muscles"; U.S. Patent Application Publication No. 20130287736A1, published October 31, 2013, entitled "Gene Therapy for Neurodegenerative Disorders"; U.S. Patent Application Publication No. 20150182637A1, published July 2, 2015, entitled "Widespread Gene Delivery of Gene Therapy Vectors"; U.S. Patent Application Publication No. 20150259391A1, published September 17, 2015, entitled "Treatment of Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No. 20160272976A1, published September 22, 2016, entitled "Products and Methods for Treatment of Familial Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No. 20160130567A1, published May 12, 2016, entitled "Messenger UNA Molecules and Uses Thereof"; U.S. Patent Application Publication No. 20180021364A1, published January 25, 2018, entitled "Central Nervous System Targeting Polynucleotides"; U.S. Patent Application Publication No. 20200297868A1, published September 24, 2020, entitled "Methods and compositions for the treatment of ALS"; U.S. Patent Application Publication No., published November 15, 2018, entitled "Translatable molecules and synthesis thereof"US Patent Application Publication No. 20180327471A1; US Patent Application Publication No. 20200247854A1 published on August 6, 2020, entitled "Pharmaceutical composition for preventing or treating neurodegenerative disease comprising nckap1 protein or gene encoding same"; US Patent Application Publication No. 20210024907A1 published on January 28, 2021, entitled "Nucleic acid-based therapeutics"; US Patent Application Publication No. 20210254103A1 published on August 19, 2021, entitled "Treatment of amyotrophic lateral sclerosis and disorders associated with the spinal cord"; US Patent Application Publication No. 20200172590A1 published on June 4, 2020, entitled "Methods of treating neurological diseases"; US Patent Application Publication No. 20220090036A1 published on March 24, 2022, entitled "Compositions and methods for the targeting of SOD1"; International Patent Application Publication No. WO2021205010A1 published on October 14, 2021, entitled "Nucleic acids encoding human FUS protein and use in the treatment of amyotrophic lateral sclerosis (ALS)"; International Patent Application Publication No. WO2022060857A1 published on March 24, 2022, entitled "Compositions and methods for treating amyotrophic lateral sclerosis (ALS) with aav-miR-SOD1"; The entire contents of each of these are hereby incorporated by reference into this specification.

[0314] Certain gene therapies provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the SOD1 gene / gene product, such as the C9orf72, ATXN2, and / or FUS gene / gene product.

[0315] Molecular payload targeting ATXN2 The ATXN2 gene encoding the ataxin-2 protein and mutations therein are involved in ALS, which affects predominantly upper and lower motor neurons. Thus, in some embodiments, modulation of ATXN2 expression and activity (by way of example, suppressing the expression and / or activity of mutant ATXN2 protein and / or its interaction with other proteins) may have a therapeutic effect in subjects having ALS.

[0316] Oligonucleotide In some embodiments, the expression and / or activity of ATXN2 (and / or the ataxin-2 protein encoded by ATXN2) may be modulated by use of an oligonucleotide that targets the ATXN2 sequence.

[0317] In some embodiments, an oligonucleotide useful for treating ALS that targets ATXN2 (by way of example, directly or indirectly modulating its expression or activity) comprises a region of complementarity to an ATXN2 transcript provided in Table 3 provided by any one of SEQ ID NOs: 396-400.

[0318] In some embodiments, examples of oligonucleotides that target ATXN2 (e.g., that directly or indirectly modulate its expression or activity) and are useful for the treatment of ALS are provided below: Becker et al. (2017) “Therapeutic reduction of ataxin-2 extends lifespan and reduces pathology in TDP-43 mice” Nature 544:367-371; Scoles et al. (2017) “Antisense oligonucleotide therapy for spinocerebellar ataxia type 2” Nature 544:362-366; US Patent Application Publication No. 20110142789A1, published June 16, 2011, entitled “Compositions and Methods for the Diagnosis and Treatment of Amyotrophic Lateral Sclerosis”; US Patent Application Publication No. 20130225659A1, published August 29, 2013, entitled “Modulation of nuclear-retained RNA”; US Patent Publication No. 1107486B2, published August 3, 2021, entitled “Compounds and methods for reducing ATXN2 expression”; US Patent Application Publication No. 20220064639A1, published March 3, 2022, entitled “Compounds and methods for reducing ATXN2 expression”; US Patent Publication No. 10533178B2, published January 14, 2020, entitled “Methods for modulating Ataxin 2 expression”; US Patent Publication No. 10006027B2, published June 26, 2018, entitled “Methods for modulating Ataxin 2 expression”; US Patent Publication No., published June 4, 2019, entitled “Compositions for modulating Ataxin 2 expression”10308934B2; US Patent Publication No. 11111494B2, published September 7, 2021, entitled "Compositions for modulating Ataxin 2 expression"; US Patent Publication No. 11345915B2, published May 31, 2022, entitled "RNA modulating oligonucleotides with improved characteristics for the treatment of neuromuscular disorders"; US Patent Application Publication No. 20140378533A1, published December 25, 2014, entitled "Modulation of RNA by repeat targeting"; US Patent Application Publication No. 20150148404A1, published May 28, 2015, entitled "RNA Modulating Oligonucleotides with Improved Characteristics for the Treatment of Neuromuscular Disorders"; US Patent Application Publication No. 20210169914A1, published June 10, 2021, entitled "Nucleic acids and nucleic acid analogs for treating, preventing, and disrupting pathological polynucleotide-binding protein inclusions"; US Patent Application Publication No. 20160040163A1, published February 11, 2016, entitled "DNAi for the modulation of genes"; US Patent Publication No. 10174328B2, published January 8, 2019, entitled "Compositions and methods for treating amyotrophic lateral sclerosis"; US Patent Application Publication No. 20220162615A1, published May 26, 2022, entitled "Methods for reducing ataxin-2 expression"; the entire contents of each of these are hereby incorporated by reference herein.

[0319] Certain oligonucleotides provided in this section may be useful in treating ALS by modulating the activity of genes and / or gene products other than the ATXN2 gene / gene product, such as the C9orf72, SOD1, and / or FUS gene / gene product.

[0320] Polypeptide In some embodiments, ATXN2 expression and / or activity may be modulated by the use of a polypeptide that can interact with ataxin-2 or an ataxin-2 polypeptide (e.g., to modulate its biological activity and / or its interaction with other biomolecules).

[0321] In some embodiments, examples of polypeptides useful for the treatment of ALS (e.g., peptides; proteins such as enzymes; antibodies, etc.) are provided below: U.S. Patent Application Publication No. 20110142789A1, published June 16, 2011, entitled "Compositions and Methods for the Diagnosis and Treatment of Amyotrophic Lateral Sclerosis"; U.S. Patent Publication No. 10066007B2, published September 4, 2019, entitled "Dipeptide-repeat proteins as therapeutic target in neurodegenerative diseases with hexanucleotide repeat expansion"; U.S. Patent Publication No. 11273149B2, published March 15, 2022, entitled "Compositions and methods for the treatment of amyotrophic lateral sclerosis, Parkinson’s disease, Parkinson’s disease with dementia, dementia with Lewy bodies, and multiple system atrophy"; U.S. Patent Publication No. 8673852B2, entitled "Methods of treating neuronal disorders using MNTF peptides and analogs thereof"; International PCT Application Publication No. WO2021222168A2, published November 4, 2021, entitled "Compositions and methods for the treatment of tdp-43 proteinopathies"; the entire contents of each of these are incorporated herein by reference.

[0322] Certain polypeptides provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the ATXN2 gene / gene product, such as the C9orf72, SOD1, and / or FUS gene / gene product.

[0323] Small molecule In some embodiments, ATXN2 expression and / or activity can be modulated by the use of small molecules that can modulate ATXN2 (e.g., to regulate its biological activity, its expression, and / or its interaction with other biomolecules).

[0324] In some embodiments, examples of small molecules useful for the treatment of ALS are provided in: U.S. Patent Application Publication No. 20110142789A1, published June 16, 2011, entitled "Compositions and Methods for the Diagnosis and Treatment of Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No. 20130303562A1, published November 14, 2013, entitled "Chemical and RNAi suppressors of neurotoxicity in Huntington’s disease"; U.S. Patent Application Publication No. 20140228333A1, published March 29, 2016, entitled "Methods for inhibiting muscle atrophy"; International PCT Application Publication No. WO2021174167A1, published September 2, 2021, entitled "Compounds and methods for modulating splicing"; International PCT Patent Application Publication No. WO2013043669A1, published March 28, 2013, entitled "Peptoid compositions for the treatment of Alzheimer’s disease and polyglutamine expansion disorder"; U.S. Patent Publication No. 10159670B2, published December 25, 2018, entitled "Methods of diagnosing and treating motor neuron diseases and other cellular stress-related diseases"; U.S. Patent Publication No. 9790188B2, published October 17, 2017, entitled "Benzimidazole derivatives and uses thereof"; the entire contents of each of which are incorporated herein by reference.

[0325] In some embodiments, the small molecule is a pharmaceutically acceptable salt, co-crystal, tautomer, stereoisomer, solvate, hydrate, polymorph, isotope-enriched compound, or prodrug of the small molecules provided herein.

[0326] Certain small molecules provided in this section may be useful in treating ALS by modulating the activity of genes and gene products other than the ATXN2 gene / gene product, such as the C9orf72, SOD1, and / or FUS gene / gene product.

[0327] Gene therapy In some embodiments, ATXN2 expression and / or activity can be modulated by the use of gene therapy that can modulate ATXN2 (e.g., by delivery of a nucleic acid encoding ATXN2, or another molecule that interacts with ATXN2).

[0328] In some embodiments, gene therapies useful in treating ALS, such as those involving administration of a compound encoding a useful therapeutic agent, are provided below: U.S. Patent Application Publication No. 20110142789A1, published on June 16, 2011, entitled "Compositions and Methods for the Diagnosis and Treatment of Amyotrophic Lateral Sclerosis"; U.S. Patent Application Publication No. 20100047261A1, published on February 25, 2010, entitled "Base-modified RNA for increasing the expression of a protein"; U.S. Patent Application Publication No. 20100203076A1, published on August 12, 2010, entitled "Complexes of RNA and cationic peptides for transfection and for immunostimulation"; U.S. Patent Publication No. 10815463B2, published on October 27, 2020, entitled "Messenger UNA molecules and uses thereof"; U.S. Patent Publication No. 11155817B2, published on October 26, 2021, entitled "Therapeutic for treatment of diseases including the central nervous system"; U.S. Patent Application Publication No. 20180327471A1, published on November 15, 2018, entitled "Translatable molecules and synthesis thereof"; the entire contents of each of these are hereby incorporated by reference herein.

[0329] Certain gene therapies provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the ATXN2 gene / gene product, such as the C9orf72, SOD1, and / or FUS gene / gene product.

[0330] Molecular payload targeting C9orf72 The C9orf72 gene encoding the chromosome 9 open reading frame 72 protein and mutations therein are involved in ALS, which predominantly affects upper and lower motor neurons. Thus, in some aspects, modulation of C9orf72 expression and activity (e.g., by suppressing the expression of mutant C9orf72 and / or the activity of the protein encoded thereby) may have a therapeutic effect in subjects having ALS.

[0331] Oligonucleotide In some aspects, C9orf72 expression and / or activity can be modulated by use of an oligonucleotide that targets the C9orf72 sequence.

[0332] In some aspects, oligonucleotides useful for the treatment of ALS that target C9orf72 (e.g., that directly or indirectly modulate its expression or activity) include regions of complementarity to the C9orf72 transcripts provided in Table 3 provided by any one of SEQ ID NOs: 393 - 395.

[0333] In some embodiments, examples of oligonucleotides useful for the treatment of ALS that target C9orf72 (e.g., directly or indirectly regulate its expression or activity) are provided below: U.S. Patent Publication No. 10577604B2, published March 3, 2020, entitled "Methods for monitoring C9ORF72 expression"; U.S. Patent Publication No. 10443052B2, published October 15, 2019, entitled "Compositions for modulating C9ORF72 expression"; U.S. Patent Publication No. 10793855B2, published October 6, 2020, entitled "Compositions for modulating expression of C9ORF72 antisense transcript"; U.S. Patent Publication No. 10815483B2, published October 27, 2020, entitled "Compositions for modulating C9ORF72 expression"; U.S. Patent Publication No. 11260073B2, published March 1, 2022, entitled "Compositions and methods for modulating C9ORF72"; U.S. Patent Publication No. 10407678B2, published October 9, 2019, entitled "Compositions for modulating expression of C9ORF72 antisense transcript"; U.S. Patent Publication No. 11162096B2, published November 2, 2021, entitled "Compositions for modulating expression of C9ORF72 antisense transcript"; U.S. Patent Publication No. US10066228B2, published September 4, 2018, entitled "Oligonucleotides for treating expanded repeat diseases"; U.S. Patent Publication No., published May 31, 2022, entitled "RNA modulating oligonucleotides with improved characteristics for the treatment of neuromuscular disorders"US Patent No. 11345915B2; US Patent Publication No. 9963699B2, published May 8, 2018, titled "Methods for modulating C9ORF72 expression"; US Patent Publication No. 10221414B2, published March 5, 2019, titled "Compositions for modulating C9ORF72 expression"; US Patent Application Publication No. 20160108396A1, published April 21, 2016, titled "Oligomers targeting hexanucleotide repeat expansion in human C9ORF72 gene"; US Patent Publication No. 10538762, published January 21, 2020, titled "Allele selective inhibition of mutant C9orf72 foci expression by duplex RNAS targeting the expanded hexanucleotide repeat"; US Patent Publication No. 10597660B2, published March 24, 2020, titled "Compositions and methods of treating amyotrophic lateral sclerosis (ALS)"; US Patent Publication No. 11118179B2, published September 14, 2021, titled "Mixed tricyclo-DNA, 2’-modified RNA oligonucleotide compositions and uses thereof"; US Patent Application Publication No. 20210284629A1, published September 16, 2021, titled "Methods and compounds for the treatment of genetic disease"; US Patent Application Publication No., published September 2, 2021, titled "Compositions and methods for reducing spliceopathy and treating rna dominance disorders"20210269825A1; US Patent Application Publication No. 20200385737A1, published on December 10, 2020, entitled "OLIGONUCLEOTIDE - OLIGONUCLEOTIDE - BASED MODULATION OF C9orf72"; US Patent Application Publication No. 20200385723A1, published on December 10, 2020, entitled "Anti - c9orf72 oligonucleotides and related methods"; US Patent Application Publication No. 20220145300A1, published on May 12, 2022, entitled "Oligonucleotide compositions and methods of use thereof"; US Patent Application Publication No. 20210032620A1, published on February 4, 2021, entitled "Oligonucleotide compositions and methods thereof"; International PCT Application Publication No. WO2021119226A1, published on December 10, 2020, entitled "Human chromosome 9 open reading frame 72 (c9orf72) irna agent compositions and methods of use thereof"; US Patent Application Publication No. 20210340535A1, published on November 4, 2021, entitled "DUAL - ACTING siRNA BASED MODULATION OF C9orf72"; International PCT Application Publication No. WO2021205005A2, published on October 14, 2021, entitled "Antisense sequences for treating amyotrophic lateral sclerosis"; the entire contents of each of these are hereby incorporated by reference into this specification.

[0334] Certain oligonucleotides provided in this section may be useful for treating ALS by modulating the activity of genes and / or gene products other than the C9orf72 gene / gene product, such as the ATXN2, SOD1, and / or FUS genes / gene products.

[0335] Polypeptide In some embodiments, the expression of C9orf72 and / or the activity of the protein encoded thereby can be regulated by the use of polypeptides, such as polypeptides that can interact with C9orf72 and / or the protein encoded thereby (e.g., to regulate its biological activity and / or its interaction with other biomolecules).

[0336] Examples of polypeptides (e.g., peptides; proteins such as enzymes; antibodies, etc.) useful for the treatment of ALS in some embodiments are provided in: U.S. Patent Publication No. 10295547B2, published May 21, 2019, entitled "Use and treatment of di-amino acid repeat-containing proteins associated with ALS"; U.S. Patent Publication No. 11197911B2, published December 14, 2021, entitled "Peptidylic inhibitors targeting C9ORF72 hexanucleotide repeat-mediated neurodegeneration"; U.S. Patent Application Publication No. 20220153874A1, published May 19, 2022, entitled "Human-derived anti-(poly-ga) dipeptide repeat (dpr) antibody"; U.S. Patent Publication No. 9329182B2, published May 3, 2016, entitled "Method of treating motor neuron disease with an antibody that agonizes MuSK"; the entire contents of each of these are incorporated herein by reference.

[0337] Certain polypeptides provided in this section may be useful for treating ALS by regulating the activity of genes and gene products other than the C9orf72 gene / gene product, such as the ATXN2, SOD1, and / or FUS gene / gene product.

[0338] Small molecule In some embodiments, C9orf72 expression and / or the activity of the protein it encodes can be modulated by the use of small molecules that can modulate C9orf72 (e.g., to regulate its biological activity, its expression, and / or its interaction with other biomolecules).

[0339] In some embodiments, examples of small molecules useful for the treatment of ALS are provided in: U.S. Patent Publication No. 10675293B2, published June 9, 2020, entitled "Nucleoside agents for the reduction of the deleterious activity of extended nucleotide repeat containing genes"; International PCT Application Publication No. WO2021174167A1, published September 2, 2021, entitled "Compounds and methods for modulating splicing"; U.S. Patent Publication No. 11241417B2, published February 8, 2022, entitled "Compositions and methods for the treatment and prevention of neurological disorders"; the entire contents of each of these are incorporated herein by reference.

[0340] In some embodiments, the small molecule is a pharmaceutically acceptable salt, co-crystal, tautomer, stereoisomer, solvate, hydrate, polymorph, isotopically enriched compound, or prodrug of the small molecule provided herein.

[0341] Certain small molecules provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the C9orf72 gene / gene product, such as the ATXN2, SOD1, and / or FUS gene / gene product.

[0342] Gene Therapy In some embodiments, C9orf72 expression and / or activity can be modulated by the use of gene therapy that can regulate C9orf72 (e.g., by delivery of nucleic acids encoding C9orf72, or other molecules that interact with the protein it encodes).

[0343] In some embodiments, gene therapies useful for the treatment of ALS, e.g., those involving administration of compounds encoding useful therapeutic agents, are provided in: U.S. Patent Publication No. 10597660B2, published Mar. 24, 2020, entitled “Compositions and methods of treating amyotrophic lateral sclerosis (ALS)”; U.S. Patent Application Publication No. 20210269825A1, published Sep. 2, 2021, entitled “Compositions and methods for reducing spliceopathy and treating rna dominance disorders”; U.S. Patent Publication No. 10801027B2, published Oct. 13, 2020, entitled “Inhibitors of SRSF1 to treat neurodegenerative disorders”; International PCT Application Publication No. WO2021160464A1, published Aug. 19, 2021, entitled “Gene therapy”; the entire contents of each of these are incorporated herein by reference.

[0344] Certain gene therapies provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the C9orf72 gene / gene product, e.g., the ATXN2, SOD1, and / or FUS gene / gene product.

[0345] Molecular payload targeting FUS The FUS gene encoding the RNA-binding protein FUS / TLS and mutations therein are involved in ALS, which affects predominantly upper and lower motor neurons. Thus, in some aspects, modulation of FUS expression and activity (e.g., by suppressing the expression of mutant FUS and / or the activity of the protein encoded thereby) may have a therapeutic effect in subjects having ALS.

[0346] Oligonucleotide In some aspects, FUS expression and / or activity can be modulated by use of oligonucleotides that target the FUS sequence.

[0347] In some aspects, oligonucleotides useful for the treatment of ALS, e.g., targeting FUS (e.g., directly or indirectly modulating its expression or activity), include regions of complementarity to the FUS transcript provided in Table 3 provided by any one of SEQ ID NOs: 401-404.

[0348] In some embodiments, examples of oligonucleotides useful for the treatment of ALS that target FUS (e.g., directly or indirectly modulate its expression or activity) are provided below: International PCT Application Publication No. WO2020243292A1, published December 3, 2020, entitled "Compounds and methods for reducing fus expression"; US Patent Publication No. 11332733B2, published May 17, 2022, entitled "Modified compounds and uses thereof"; US Patent Application Publication No. 20100256223A1, published October 7, 2010, entitled "Oligonucleotides for modulating target rna activity"; US Patent Publication No. 9150860B2, published October 6, 2015, entitled "FUS / TLS-based compounds and methods for diagnosis, treatment and prevention of amyotrophic lateral sclerosis and related motor neuron diseases"; US Patent Application Publication No. 20120252875A1, published October 4, 2012, entitled "Methods and compositions for treating diseases, disorders or injury of the CNS"; US Patent Publication No. 10781445B2, published September 22, 2020, entitled "Decoy oligonucleotides for the treatment of diseases"; US Patent Application Publication No. 20190127733A1, published May 2, 2019, entitled "Oligonucleotide compositions and methods thereof"; US Patent Publication No., published December 14, 2021, entitled "Inhibition of stress granule formation through manipulation of UBAP2L"11197883B2; U.S. Patent Application Publication No. 20210169914A1, published on June 10, 2021, entitled "Nucleic acids and nucleic acid analogs for treating, preventing, and disrupting pathological polynucleotide-binding protein inclusions"; International PCT Application Publication No. WO2021203043A2, published on October 7, 2021, entitled "Targeted inhibition using engineered oligonucleotides"; International PCT Application Publication No. WO2021207854A1, published on October 21, 2021, entitled "Compositions and methods for inhibiting tdp-43 and fus aggregation"; the entire contents of each of these are incorporated herein by reference.

[0349] Certain oligonucleotides provided in this section may be useful for treating ALS by modulating the activity of genes and / or gene products other than the FUS gene / gene product, such as the ATXN2, SOD1, and / or C9orf72 gene / gene product.

[0350] Polypeptide In some embodiments, FUS expression and / or the activity of the protein encoded thereby can be modulated by the use of polypeptides, such as polypeptides that can interact with the FUS nucleic acid and / or the protein encoded thereby (e.g., to modulate its biological activity, its intracellular localization, and / or its interaction with other biomolecules).

[0351] In some embodiments, examples of polypeptides (such as peptides; proteins such as enzymes; antibodies, etc.) useful for the treatment of ALS are provided below: U.S. Patent Publication No. 1132504B2, published May 17, 2022, entitled "Methods of reducing FUS / TLS- or TDP-43-mediated neuronal cytotoxicity by UPF1"; U.S. Patent Application Publication No. 20180360925A1, published December 20, 2018, entitled "Extracellular dna as a therapeutic target in neurodegeneration"; the entire contents of each of these are incorporated herein by reference.

[0352] Certain polypeptides provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the FUS gene / gene product, such as the ATXN2, SOD1, and / or C9orf72 gene / gene product.

[0353] Small molecule In some embodiments, FUS expression and / or the activity of the protein it encodes can be modulated by the use of small molecules that can modulate FUS (e.g., to modulate its biological activity, its expression, its localization, and / or its interaction with other biomolecules).

[0354] In some embodiments, examples of small molecules useful for the treatment of ALS are provided below: US Patent Application Publication No. 20120272345A1, published on October 25, 2012, entitled "Diagnosis marker, diagnosis method and therapeutic agent for amyotrophic lateral sclerosis, and animal model and cell model developing amyotrophic lateral sclerosis"; US Patent Publication No. 10159670B2, published on December 25, 2018, entitled "Methods of diagnosing and treating motor neuron diseases and other cellular stress-related diseases"; US Patent Application Publication No. 20200216563A1, published on July 9, 2020, entitled "Hdac6 and protein aggregation"; International PCT Application Publication No. WO2021174167A1, published on September 2, 2021, entitled "Compounds and methods for modulating splicing"; US Patent Application Publication No. 20200368267A1, published on November 26, 2020, entitled "Prophylactic and / or therapeutic agent for amyotrophic lateral sclerosis"; US Patent Application Publication No. 20220071955A1, published on March 10, 2022, entitled "Methods of Treatment, Prevention and Diagnosis"; the entire contents of each of these are incorporated herein by reference.

[0355] In some embodiments, the small molecule is a pharmaceutically acceptable salt, co-crystal, tautomer, stereoisomer, solvate, hydrate, polymorph, isotope-enriched compound, or prodrug of the small molecule provided herein.

[0356] Certain small molecules provided in this section may be useful in treating ALS by modulating the activity of genes and gene products other than the FUS gene / gene product, such as the ATXN2, SOD1, and / or C9orf72 gene / gene product.

[0357] Gene therapy In some embodiments, FUS expression and / or activity can be modulated by the use of gene therapy that can modulate FUS, e.g., by delivery of nucleic acids encoding FUS, or an FUS transcript or other molecule that interacts with the protein encoded by FUS.

[0358] In some embodiments, gene therapies useful in the treatment of ALS, e.g., those involving administration of a compound encoding a useful therapeutic agent, are provided in International PCT Application Publication No. WO2021205010A1, published October 14, 2021, entitled "Nucleic acids encoding human FUS protein and use in the treatment of amyotrophic lateral sclerosis (ALS)"; the entire contents of each of which are incorporated herein by reference.

[0359] Certain gene therapies provided in this section may be useful in treating ALS by modulating the activity of genes and gene products other than the FUS gene / gene product, such as the ATXN2, SOD1, and / or C9orf72 gene / gene product.

[0360] Molecular payload targeting PIKFYVE The PIKFYVE gene encoding phosphatidylinositol-3-phosphate 5-kinase type III (PIPKIII) protein and mutations therein are involved in ALS. Thus, in some aspects, modulation of PIKFYVE expression and activity (e.g., by suppressing the expression and / or activity of mutant PIPKIII protein and / or its interaction with other proteins) may have a therapeutic effect in subjects with ALS.

[0361] Oligonucleotide In some aspects, the expression and / or activity of PIKFYVE (and / or the PIPKIII protein encoded by PIKFYVE) can be modulated by the use of oligonucleotides that target the PIKFYVE sequence.

[0362] In some aspects, oligonucleotides useful for the treatment of ALS that target PIKFYVE (e.g., that directly or indirectly modulate its expression or activity) include regions of complementarity to the PIKFYVE transcripts provided in Table 3 provided by any one of SEQ ID NOs: 143-148.

[0363] In some aspects, examples of oligonucleotides useful for the treatment of ALS that target PIKFYVE (e.g., that directly or indirectly modulate its expression or activity) are provided in US20220411804A1 published December 29, 2022, entitled "Pikfyve antisense oligonucleotides"; the entire content of which is incorporated herein by reference.

[0364] Certain oligonucleotides provided in this section may be useful for treating ALS by modulating the activity of genes and / or gene products other than the PIKFYVE gene / gene product, e.g., other genes / gene products associated with ALS.

[0365] Polypeptide In some embodiments, PIKFYVE expression and / or activity can be regulated by the use of a polypeptide that can interact with PIPKIII (e.g., to modulate its biological activity and / or its interaction with other biomolecules) or a PIPKIII polypeptide.

[0366] In some embodiments, examples of polypeptides useful for the treatment of ALS (e.g., peptides; proteins such as enzymes; antibodies, etc.) include PIPKIII proteins and functional fragments thereof.

[0367] Certain polypeptides provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the PIKFYVE gene / gene product, e.g., other genes / gene products associated with ALS.

[0368] Small molecule In some embodiments, PIKFYVE expression and / or activity can be regulated by the use of small molecules that can regulate PIPKIII (e.g., to modulate its biological activity, its expression, and / or its interaction with other biomolecules).

[0369] In some embodiments, examples of small molecules useful for the treatment of ALS are provided below: US20190192527A1, published June 27, 2019, entitled "Compositions comprising pikfyve inhibitors and methods related to inhibition of rank signaling"; WO2017040971A1, published March 9, 2017, entitled "Methods of using inhibitors of pikfyve for the treatment of lysosomal storage disorders and neurodegenerative diseases"; WO2022086993A1, published April 28, 2022, entitled "Novel inhibitors of pikfyve and methods using same"; US20210139505A1, published May 13, 2021, entitled "PIKfyve Inhibitors"; US10758545B2, published September 1, 2020, entitled "Methods to treat neurological diseases"; US11066410B2, published July 20, 2021, entitled "Fused triazolo-pyrimidine compounds having useful pharmaceutical application"; the entire contents of each of these are incorporated herein by reference.

[0370] In some embodiments, the small molecule is a pharmaceutically acceptable salt, co-crystal, tautomer, stereoisomer, solvate, hydrate, polymorph, isotopically enriched compound, or prodrug of the small molecule provided herein.

[0371] Certain small molecules provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the PIKFYVE gene / gene product, such as other genes / gene products associated with ALS.

[0372] Gene therapy In some embodiments, PIKFYVE expression and / or activity can be regulated by the use of gene therapy that can modulate PIKFYVE, for example, by delivery of nucleic acids encoding PIKFYVE, or other molecules that interact with PIKFYVE.

[0373] In some embodiments, gene therapies useful for the treatment of ALS, for example, those involving administration of compounds encoding useful therapeutic agents, include payloads encoding PIPKIII or functional fragments thereof.

[0374] Certain gene therapies provided in this section may be useful for treating ALS by modulating the activity of genes and gene products other than the PIKFYVE gene / gene product, for example, other genes / gene products associated with ALS.

[0375] Molecular payload targeting SYF2 The SYF2 gene encoding the pre-mRNA splicing factor SYF2 protein and mutations therein are involved in ALS. Thus, in some embodiments, modulation of SYF2 expression and activity, for example, by suppressing the expression and / or activity of mutant pre-mRNA splicing factor SYF2 protein and / or its interaction with other proteins, may have a therapeutic effect in subjects with ALS.

[0376] Oligonucleotide In some embodiments, the expression and / or activity of SYF2 (and / or the pre-mRNA splicing factor SYF2 protein encoded by SYF2) can be regulated by the use of oligonucleotides that target the SYF2 sequence.

[0377] In some embodiments, oligonucleotides useful for the treatment of ALS that target, e.g., SYF2 (e.g., directly or indirectly modulate its expression or activity) include regions of complementarity to SYF2 transcripts provided in Table 3 provided by any one of SEQ ID NOs: 167-168.

[0378] In some embodiments, examples of oligonucleotides useful for the treatment of ALS that target, e.g., SYF2 (e.g., directly or indirectly modulate its expression or activity) are provided in US20230066380A1, published March 2, 2023, entitled "Antagonism as a therapy for tdp-43 proteinopathies"; the entire content of which is incorporated herein by reference.

[0379] Certain oligonucleotides provided in this section may be useful for treating ALS by modulating the activity of genes and / or gene products other than the SYF2 gene / gene product, e.g., other genes / gene products associated with ALS.

[0380] Polypeptide In some embodiments, SYF2 expression and / or activity can be modulated by the use of a polypeptide that can interact with the pre-mRNA splicing factor SYF2 (e.g., to modulate its biological activity and / or its interaction with other biomolecules) or the pre-mRNA splicing factor SYF2 polypeptide.

[0381] In some embodiments, examples of polypeptides useful for the treatment of ALS (e.g., peptides; proteins such as enzymes; antibodies, etc.) include the pre-mRNA splicing factor SYF2 protein and functional fragments thereof.

[0382] Certain polypeptides provided in this section may be useful in treating ALS by modulating the activity of genes and gene products other than the SYF2 gene / gene product, such as other genes / gene products associated with ALS.

[0383] Small molecule In some embodiments, SYF2 expression and / or activity can be modulated by the use of small molecules that can modulate the pre-mRNA splicing factor SYF2 protein (e.g., to modulate its biological activity, its expression, and / or its interaction with other biomolecules).

[0384] In some embodiments, examples of small molecules useful in treating ALS are small molecules that increase or decrease the expression of SYF2 and / or increase or decrease the level or activity of the pre-mRNA splicing factor SYF2 protein.

[0385] Certain small molecules provided in this section may be useful in treating ALS by modulating the activity of genes and gene products other than the SYF2 gene / gene product, such as other genes / gene products associated with ALS.

[0386] Gene therapy In some embodiments, SYF2 expression and / or activity can be modulated by the use of gene therapy that can modulate SYF2 (e.g., by deliver...

Claims

1. A complex comprising an anti-TfR1 antibody covalently linked to a molecular payload for treating a central nervous system (CNS) disease or disorder, wherein the anti-TfR1 antibody is (i) heavy chain complementarity determining region 1 (CDR-H1) of SEQ ID NO: 1, heavy chain complementarity determining region 2 (CDR-H2) of SEQ ID NO: 2, heavy chain complementarity determining region 3 (CDR-H3) of SEQ ID NO: 3, light chain complementarity determining region 1 (CDR-L1) of SEQ ID NO: 4, light chain complementarity determining region 2 (CDR-L2) of SEQ ID NO: 5, and light chain complementarity determining region 3 (CDR-L3) of SEQ ID NO: 6; (ii) CDR-H1 of SEQ ID NO: 7, CDR-H2 of SEQ ID NO: 8, CDR-H3 of SEQ ID NO: 9, CDR-L1 of SEQ ID NO: 10, CDR-L2 of SEQ ID NO: 11, and CDR-L3 of SEQ ID NO: 6; or (iii) CDR-H1 of SEQ ID NO: 12, CDR-H2 of SEQ ID NO: 13, CDR-H3 of SEQ ID NO: 14, CDR-L1 of SEQ ID NO: 15, CDR-L2 of SEQ ID NO: 5, and CDR-L3 of SEQ ID NO: 16; wherein the complex delivers the molecular payload to cells of the CNS comprising the said complex.

2. The complex according to claim 1, wherein the anti-TfR1 antibody comprises a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 17 and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO:

18.

3. The complex according to claim 1 or claim 2, wherein the anti-TfR1 antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 19 and a light chain comprising the amino acid sequence of SEQ ID NO:

20.

4. The complex according to any one of claims 1 to 3, wherein the anti-TfR1 antibody is a Fab.

5. The complex according to any one of claims 1 to 4, wherein the molecular payload is configured to regulate the expression of a gene associated with a CNS disease or disorder.

6. The complex according to any one of claims 1 to 5, wherein the molecular payload comprises an oligonucleotide, a polypeptide, a small molecule, or a gene therapy payload, optionally wherein the gene therapy payload comprises a messenger RNA (mRNA) molecule.

7. The complex according to any one of claims 1 to 6, wherein the anti-TfR1 antibody is covalently linked to the molecular payload via a linker comprising the structure of formula (I): 【Chemical 1】 wherein n is any number from 0 to 10, m is any number from 0 to 10, optionally n is 3, and / or m is 4; L1 is a spacer which is a substituted or unsubstituted aliphatic, substituted or unsubstituted heteroaliphatic, substituted or unsubstituted carbocyclylene, substituted or unsubstituted heterocyclylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, -O-, -N(R A )-, -S-, -C(=O)-, -C(=O)O-, -C(=O)NR A -, -NR A C(=O)-, -NR A C(=O)R A -, -C(=O)R A -, -NR A C(=O)O-, -NR A C(=O)N(R A )-, -OC(=O)-, -OC(=O)O-, -OC(=O)N(R A )-, -S(O) 2 NR A -, -NR A S(O) 2 -, or a combination thereof, wherein each R A is independently hydrogen or substituted or unsubstituted alkyl.

8. The complex according to any one of claims 1 to 7, wherein the complex comprises the structure of formula (J): 【Chemical Formula 2】 wherein n is any number from 0 to 10, m is any number from 0 to 10, optionally, n is 3, and / or m is 4. **Claim 9** The complex according to any one of claims 1 to 8, wherein the complex delivers a molecular payload to cells of the CNS by passing through the blood-brain barrier. **Claim 10** The complex according to any one of claims 1 to 8, wherein the complex delivers a molecular payload to cells of the CNS by passing through the choroid plexus. **Claim 11** A gene associated with a CNS disease or disorder is (i) DMPK, DMD, SMN, or FXN; (ii) SOD1, C9orf72, ATXN2, or FUS; (iii) LRRK2 or SNCA; (iv) HTT or MSH3; (v) TREM2, APOE, MAPT, or APP; (vi) GYS1, PrP, VLA-4, GFAP, UBE3A, LSD, or SCN9A; or (vii) SCN1A, SCN2A, SCN8A, SCN9A, CLN3, GRIA1, or PCDH19 The complex according to any one of claims 5 to 10. **Claim 12** The complex according to any one of claims 5 to 10, wherein a gene associated with a CNS disease or disorder is TOR1A, THAP1, ANO3, GNAL, KMT2B, GCH1, TH, SPR, TAF1, PRKRA, ATP1A3, SGCE, PNKD, PRRT2, SLC2A1, or ECHS1. **Claim 13** A gene associated with a CNS disease or disorder is (i) PIKFYVE, SYF2, or UNC13A; (ii) GRIN2A; (iii) ATXN1, ATXN2, ATXN3, or MSH3; (iv) GRN, C9orf72, MAPT, PIKFYVE, SYF2, or UNC13A; (v) TPP1 or CLN3; or (vi) APOE, SCN1A, GLB1, ASM, ARSA, GALC, HEXA, HEXB, GBA, or MECP2 The complex according to any one of claims 5 to 10. **Claim 14** The complex according to any one of claims 1 to 13, wherein the molecular payload comprises an oligonucleotide comprising a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 392 to 702 or to a target sequence of an oligonucleotide listed in any one of Tables 5 to 19, and optionally, wherein the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 to 19.

15. The complex according to any one of claims 1 to 13, wherein the molecular payload comprises an oligonucleotide comprising a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 705 to 803 or to a target sequence of an oligonucleotide listed in any one of Tables 5 to 19, and optionally, wherein the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 to 19.

16. The complex according to any one of claims 1 to 13, wherein the molecular payload comprises an oligonucleotide comprising a region of complementarity to a transcript submitted with any one of SEQ ID NOs: 143 to 148, 167 to 169, 810 to 875, and 1059 to 1068 or to a target sequence of an oligonucleotide listed in any one of Tables 5 to 19, and optionally, wherein the oligonucleotide comprises an oligonucleotide structure listed in any one of Tables 5 to 19.

17. The complex according to any one of claims 1 to 16, wherein the CNS disease or disorder is a neuromuscular disease or disorder, and optionally, wherein the neuromuscular disease or disorder is Duchenne muscular dystrophy, myotonic dystrophy, Friedreich's ataxia, or spinal muscular atrophy.

18. The CNS disease or disorder is (i) amyotrophic lateral sclerosis; (ii) Parkinson's disease; (iii) essential tremor; (iv) Huntington's disease; (v) Alzheimer's disease; (vi) hereditary dystonia; (vii) epilepsy; (viii) a pain disorder; or (ix) a glycogen synthesis disorder; neurodegeneration; small fiber neuropathy; phenotypes related to nociception; Alexander disease; Angelman syndrome; autism spectrum disorder; retinitis pigmentosa; isolated macular dystrophy; and / or multiple sclerosis and is the complex according to any one of claims 1 to 16:

19. The CNS disease or disorder is (i) spinocerebellar ataxia (SCA); (ii) Frontotemporal dementia (FTD); (iii) Motor neuron disease; (iv) Dravet syndrome; (v) Batten disease; (vi) GM1 gangliosidosis; (vii) Niemann-Pick type A; (viii) Metachromatic leukodystrophy; (ix) Krabbe disease; (x) Tay-Sachs; (xi) Sandhoff disease; (xii) Gaucher disease type II or III; or (xiii) Rett syndrome. The complex according to any one of claims 1 to 16, which is such.

20. The molecular payload is the molecular payload disclosed in any one of paragraphs 0216 to 1208, and optionally, where the molecular payload is paragraphs 0296 to 0299, 0404 to 0406, 0468 to 0470, 0500 to 0502, 0535 to 0539, 0601 to 0604, 0666 to 0668, 0757 to 0759, 0779 to 0781, 0896 to 0901, 0916 to 0918, 0946 to 0948, 1049 to 1056, 1070 to 1078, 1092 to 1102, 1116 to 1124, 1138 to 1141, 1155 to 1158, 1172 to 1177, and 1191 to 1193. The complex according to any one of claims 1 to 19, which is the molecular payload disclosed in any one of them.

21. A method for treating a CNS disease or disorder, comprising administering the complex according to any one of claims 1 to 20 to a subject in need thereof.

22. A method for delivering a molecular payload to the CNS of a subject, comprising administering the complex according to any one of claims 1 to 20 to the subject.

23. The method according to claim 22, wherein the complex is administered intravenously to the subject.

24. The method according to claim 22 or 23, wherein the complex is detectable in the cortex of the subject after administration.

25. The method according to any one of claims 22 to 24, wherein the complex is detectable in the cerebellum of the subject after administration.

26. The method according to any one of claims 22 to 25, wherein the complex is detectable in the deep brain tissue of the subject after administration, and optionally, where the deep brain tissue is the thalamus, caudate nucleus, and / or putamen of the subject.

27. The method according to any one of claims 22 to 26, wherein the complex is detectable in the cortical neurons, motor neurons, cerebellar cells, and / or choroid plexus cells of the subject after administration.

28. The method according to any one of claims 22 to 27, wherein the molecular payload comprises a protein, and optionally, wherein the protein is an enzyme. **Claim 29** The method according to claim 28, wherein the subject is diagnosed with or suspected of having Batten disease, GM1 gangliosidosis, Niemann-Pick type A, metachromatic leukodystrophy, Krabbe disease, Tay-Sachs, Sandhoff disease, or Gaucher disease. **Claim 30** The method according to any one of claims 22 to 27, wherein the payload comprises an oligonucleotide. **Claim 31** The method according to claim 30, wherein the subject is diagnosed with or suspected of having ALS, Angelman syndrome, Rett syndrome, Parkinson's, Lewy body dementia, Alzheimer's disease (which may or may not be associated with cerebral amyloid angiopathy (CAA) or frontotemporal dementia), epilepsy, Alexander disease, spinal muscular atrophy, Batten disease, Huntington's disease, spinocerebellar ataxia, motor neuron disease, or Dravet syndrome.

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