AUF1 compositions and methods for promoting generation of neuromuscular junctions

Abstract

The present application relates to a method of promoting regeneration of neuromuscular junctions in a subject in need thereof. This method involves administering, to a subject in need thereof, a composition comprising a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof under conditions effective to promote regeneration of neuromuscular junctions in the subject.

Description

AUF1 COMPOSITIONS AND METHODS FOR PROMOTING GENERATION OF NEUROMUSCULAR JUNCTIONS

[0001] This application claims the priority benefit of U.S. Provisional Patent Application Serial No. 63 / 637,693, filed April 23, 2024, which is hereby incorporated by reference in its entirety.

[0002] This invention was made with government support under R01 AR074430 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0003] The present disclosure relates to methods of promoting regeneration of neuromuscular junctions in a subject in need thereof by administering compositions comprising a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof.BACKGROUND

[0004] Muscle contraction is triggered by transduction of the acetylcholine signal at the neuromuscular junction (NMJ) and relies on a high density of acetylcholine receptors (AChRs) on the surface of the postsynaptic membrane in the muscle cell. AChRs are heteromeric membrane proteins that cluster to ensure proper signal transduction and are essential for neurotransmission at the neuromuscular junction. Several NMJ-associated proteins are essential for NMJ function. Rapsyn, an intracellular peripheral membrane protein that binds AChRs, is essential for synaptic differentiation. Rapsyn is crucial for anchoring AChRs in the postsynaptic membrane. Musk, another NMJ-associated protein, is essential for the formation and maintenance of NMJs and initiates postsynaptic differentiation, priming the muscle for synapse formation.

[0005] Many conditions such as traumatic injuries, aging or denervation induces destabilization of AChRs and results in muscle denervation.

[0006] There are no approved treatments to increase NMJ function or formation. Thus, there remains an urgent need for effective therapeutic options that increase NMJ function and formation.

[0007] The present disclosure is directed to overcoming these and other deficiencies in the art.SUMMARY

[0008] One aspect of the present disclosure relates to a method of promoting regeneration of neuromuscular junctions in a subject in need thereof. This method involves administering, to the subject in need thereof, a composition comprising a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof under conditions effective to promote regeneration of neuromuscular junctions in the subject.

[0009] There are currently no approved therapeutic interventions to promote regeneration of functional skeletal muscle neuro-muscular junctions (NMJs) which are essential to provide muscle function. Neuromuscular junctions (NMJs) are specialized muscle synapses that innervate and control muscle function. They are essentially composed by acetylcholine receptors (AChRs). Loss of NMJs occurs as a result traumatic injury resulting in muscle denervation, neurogenic myopathies, age related muscle loss and sarcopenia, genetic neuromuscular wasting diseases such as amyotrophic lateral sclerosis (ALS), spinal muscular atrophy (SMA), Duchenne Muscular Dystrophy (DMD), neurogenic inflammatory disorders such as Multiple Sclerosis (MS), nerve trauma, botulinum toxicity, disuse muscle atrophy, and many common and rare neuromuscular degenerative diseases and muscle wasting diseases.

[0010] The Examples of the present disclosure demonstrate that the key mRNAs that program the generation of NMJs are regulated in their stability and translation by the mRNA binding protein AUF1. The Examples of the present disclosure show that gene transfer of AUF1 to injured muscle, aging muscle, sarcopenic muscle, or muscle undergoing wasting as a result of muscular dystrophy, promotes very effective and rapid regeneration of NMJs and restores muscle function compared to untreated controls, in mouse studies.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIGs. 1 A-1D show an increase of essential components of neuromuscular junctions (acetylcholine, Rapsyn, Musk, and Gabpa receptors) mRNA induced by AUF1 supplementation. Mice at 2 months of age were prophylaxed i.v. with AAV8 or AAV8 AUF1 at 2E13 viral genome copies / kg (vg / kg), for 1 month then the Tibialis Anterior (TA) muscle was injured by direct injection of 1.2% BaCh, which causes instant necrosis of 40-50% of the TA muscle. FIG 1 A shows AChR encoded Chm mRNA levels in injured TA muscle at 7 days and 14 days in AAV control and AAV AUF1 supplemented mice. Rapsyn (FIG. IB), Musk (FIG.1C) and Gabpa (FIG. ID) mRNAs levels are also increased in injured TA muscle at day 7 and day 14 in AAV control and AAV AUF1 supplemented mice.

[0012] FIGs. 2A-2B show denervated myofibers stained with NCAM antibody in uninjured muscle, and injured muscle after 14 and 30 days post-injury. Prophylactic i.v. AAV8 or AAV8 AUF1 supplementation (at 2E13 vg / kg) strongly reduced the percent of denervated myofiber 14 days post injury (FIG. 2A). Quantification of NCAM+staining at 14 days is provided. Mean ± SEM, n=3-4 studies (FIG. 2B). *P<0.05; **P<0.01 by ANOVA-test.

[0013] FIG. 3 shows Immunofluorescence (IF) images of a-bungarotoxin stain that images AChRs in post-synaptic NMJs, and synaptophysin stain that images the presynaptic nerve sinus in normal uninjured TA muscle and BaCE injured TA muscle, with or without AAV8 or AAV8 AUF1 prophylaxis supplementation (2E13 vg / kg). Data show that AUF1 supplementation induces exact overlay of a-bungarotoxin and synaptophysin imaging. This is an established index of NMJ innervation, which is increased dramatically by AUF1 supplementation therapy.

[0014] FIGs. 4A-4B shows the increase of essential components of neuromuscular junctions (acetylcholine, Chrn, Rapsyn, Musk) mRNAs by AUF1 supplementation therapy.Mice at 3 months of age were administered intramuscular injection of AAV8 or AAV8 AUF1 24 hours after 1.2% BaCh injury of the TA muscle (at 2E11 vg / kg). FIG. 4A shows AChR Chrn encoded mRNA levels in injured TA muscle at 14 days post-injury in AAV8 control and AAV8 AUF1 supplemented mice. Rapsyn and Musk (FIG. 4B) mRNA levels are also increased in uninjured and injured TA muscles at 14 days in AAV8 control and AAV8 AUF1 supplemented mice.

[0015] FIG. 5 shows denervated myofibers stained with NCAM antibody in uninjured muscle, and in injured muscle 14 days post-injury, when AAV8 or AAV8 AUF1 was administrated by TA intramuscular injection 24 hours post-injury (at 2E11 vg / kg).Quantification of NCAM+staining at 14 days post-injury is shown. Mean ± SEM, n=4. *P<0.05 by t-test.

[0016] FIG. 6 shows IF images of a-bungarotoxin that stains AChRs in the post-synaptic NMJ, and synaptophysin that stains the presynaptic nerve sinus in normal uninjured TA muscle and BaCh injured TA muscle, with AAV8 or AAV8 AUF1 supplementation by TA intramuscular injection 24 hours post-injury (at 2E11 vg / kg). Data show that AUF1 induces an exact overlay of a-bungarotoxin and synaptophysin an established index of NMJ innervation, which is increased dramatically by AUF1 supplementation therapy.

[0017] FIG. 7 shows that AUF1 supplementation protects muscle against atrophy and muscle strength loss after injury. AAV8 or AAV8 AUF1 was injected in the TA muscle 24hours post-injury of the TA muscle with BaCh (at 2E11 vg / kg). Mean ± SEM, n=5.***p<0 001 by t-test. ns means not significant. Animals were tested for rear leg grip strength.

[0018] FIG. 8 shows that supplementation of AUF1 in the C2C12 myoblast cell line in tissue culture delivered by lentivirus vector (LV) increases AChR Chrn gene expression as well as rapsyn expression, during myoblast differentiation into myofibers. Mean ± SEM, n=6. **P<0.01; ***P<0.001; ****P<0.0001 by t-test.DETAILED DESCRIPTIONDefinitions

[0019] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present disclosure herein described for which they are suitable as would be understood by a person skilled in the art.

[0020] Singular forms “a”, “an”, and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, a reference to “a method” includes one or more methods and / or steps of the type described herein and / or which will become apparent to a person of ordinary skill in the art upon reading this disclosure. In another example, reference to “a compound” includes both a single compound and a plurality of different compounds.

[0021] The term “about” or “approximately” includes being within a statistically meaningful range of a value. Such a range can be within an order of magnitude, such as within 50%, or within 20%, or within 10%, or within 5% (or any amount or range within 5-50%) of a given value or range. The allowable variation encompassed by the term “about” or “approximately” may depend on the context.

[0022] The term “and / or” as used herein means that the listed features are present, or used, individually or in combination. In effect, this term means that “at least one of’ or “one or more” of the listed features is used or present.

[0023] As will be understood by a person of ordinary skill in the art, for any and all purposes, such as in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof, as well as any value within a range. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, and so on. As a non-limiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, and so on. As will also be understood by a person of ordinary skill in the art all language such as “up to,” “at least,” and the like include thenumber recited and refer to ranges which can be subsequently broken down into subranges or specific values therein as discussed above. Finally, as will be understood by a person of ordinary skill in the art, and as discussed above, a range includes each individual value.

[0024] In understanding the scope of the present disclosure, the term “comprising” and its derivatives, as used herein, are intended to be open-ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, “including”, “involving”, “having”, and their derivatives. The term “consisting” and its derivatives, as used herein, are intended to be closed terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The term “consisting essentially of’, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps. In embodiments or claims where the term comprising (or the like) is used as the transition phrase, such embodiments can also be envisioned with replacement of the term “comprising” with the terms “consisting of’ or “consisting essentially of.” The methods, kits, systems, and / or compositions of the present disclosure can comprise, consist essentially of, or consist of, the components disclosed.

[0025] In some embodiments comprising an “additional” or “second” component, the second component as used herein is different from the other components or first component. A “third” component is different from the other, first, and second components, and further enumerated or “additional” components are similarly different.

[0026] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, some embodiments of the methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0027] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting, since the scope of the present disclosure will be limited only by the appended claims.Methods of Promoting Regeneration of Neuromuscular Junctions

[0028] One aspect of the present disclosure relates to a method of promoting regeneration of neuromuscular junctions in a subject in need thereof. This method involves administering, to the subject in need thereof, a composition comprising a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof under conditions effective to promote regeneration of neuromuscular junctions in the subject.

[0029] As used herein, the terms “promote,” “promotion,” and “promoting” refer to an increase in an activity, response, condition, or other biological parameter, including the production, presence, expression, or function of cells, biomolecules or bioactive molecules. The terms “promote,” “promotion,” and “promoting include, but are not limited to, initiation of an activity, response, or condition, as well as initiation of the production, presence, or expression of cells, biomolecules, or bioactive molecules. The terms “promote,” “promotion,” and “promoting” may also include measurably increasing an activity, response, or condition, or measurably increasing the production, presence, expression, or function of cells, biomolecules, or bioactive molecules, as compared to a native or control level.

[0030] Suitable subjects according to the present disclosure include, without limitation, mammals such as humans, non-human primates, rodents (mice or rats), cats, dogs, rabbits, horses, sheep, pigs, and cows. In some embodiments, the subject is a human subject. Exemplary human subjects include, without limitation, infants, children, adults, and elderly subjects.

[0031] Genes involved in rapid response to cell stimuli are highly regulated and typically encode mRNAs that are selectively and rapidly degraded to quickly terminate protein expression and reprogram the cell (Moore et al., “Physiological Networks and Disease Functions of RNA- binding Protein AUF1,” Wiley Interdiscip. Rev. RNA 5(4):549-64 (2014), which is hereby incorporated by reference in its entirety). These include growth factors, inflammatory cytokines (Moore et al., “Physiological Networks and Disease Functions of RNA-binding Protein AUF1,” Wiley Interdiscip Rev RNA 5(4):549-64 (2014) and Zhang et al., “Purification, Characterization, and cDNA Cloning of an AU-rich Element RNA-binding Protein, AUF1,” Mol. Cell. Biol. 13(12):7652-65 (1993), which are hereby incorporated by reference in their entirety), and tissue stem cell fate-determining mRNAs (Chenette et al., “Targeted mRNA Decay by RNA Binding Protein AUF1 Regulates Adult Muscle Stem Cell Fate, Promoting Skeletal Muscle Integrity,”Cell Rep. 16(5): 1379-90 (2016), which is hereby incorporated by reference in its entirety) that have very short half-lives of 5-30 minutes.

[0032] Short-lived mRNAs typically contain an AU-rich element (“ARE”) in the 3' untranslated region (“3'UTR”) of the mRNA, having the repeated sequence AUUUA (Moore et al., “Physiological Networks and Disease Functions of RNA-binding Protein AUF1,” Wiley Interdiscip Rev. RNA 5(4):549-64 (2014), which is hereby incorporated by reference in its entirety), which confers rapid decay. The ARE serves as a binding site for regulatory proteins known as AU-rich binding proteins (AUBPs) that control the stability and in some cases the translation of the mRNA (Moore et al., “Physiological Networks and Disease Functions of RNA- binding Protein AUF1,” Wiley Interdiscip. Rev. RNA 5(4):549-64 (2014); Zhang et al., “Purification, Characterization, and cDNA Cloning of an AU-rich Element RNA-binding Protein, AUF1,” Mol. Cell. Biol. 13(12):7652-65 (1993); and Halees et al., “ARED Organism: Expansion of ARED Reveals AU-rich Element Cluster Variations Between Human and Mouse,” Nucleic Acids Res 36(Database issue):D137-40 (2008), which are hereby incorporated by reference in their entirety).

[0033] AU-rich mRNA binding factor 1 (AUF1; HNRNPD) binds with high affinity to repeated AU-rich elements (“AREs”) located in the 3' untranslated region (“3' UTR”) found in approximately 5% of mRNAs. Although AUF1 typically targets ARE-mRNAs for rapid degradation, while not as well understood, it can oppositely stabilize and increase the translation of some ARE-mRNAs (Moore et al., “Physiological Networks and Disease Functions of RNA- Binding Protein AUF1,” Wiley Interdiscip. Rev. RNA 5(4): 549-564 (2014), which is hereby incorporated by reference in its entirety). It was previously reported that mice with AUF1 deficiency undergo an accelerated loss of muscle mass due to an inability to carry out the myogenesis program (Chenette et al., “Targeted mRNA Decay by RNA Binding Protein AUF1 Regulates Adult Muscle Stem Cell Fate, Promoting Skeletal Muscle Integrity,” Cell Rep.16(5): 1379-90 (2016), which is hereby incorporated by reference in its entirety). It was also found that AUF1 expression is severely reduced with age in skeletal muscle, and this significantly contributes to loss and atrophy of muscle, loss of muscle mass, and reduced strength (Abbadi et al., “Muscle Development and Regeneration Controlled by AUFl-mediated Stage-specific Degradation of Fate-determining Checkpoint mRNAs,” Proc. Natl. Acad. Sci. USA 116(23): 11285-11290 (2019), which is hereby incorporated by reference in its entirety). It was also found that AUF1 controls all major stages of skeletal muscle development, starting with satellite cell programming once activated and lineage commitment, by selectively targeting for rapid degradation the major differentiation checkpoint mRNAs that block entry into each nextstep of muscle development and promoting the stabilization and increased translation of pro- myogenic ARE-mRNAs (Abbadi et al., “Muscle Development and Regeneration Controlled by AUFl-Mediated Stage-Specific Degradation of Fate-Determining Checkpoint mRNAs,” Proc. Natl. Acad. Sci. USA 116(23): 11285-11290 (2019); Abbadi et al., “AUF1 Gene Transfer Increases Exercise Performance and Improves Skeletal Muscle Deficit in Adult Mice”, Molecular Therapy 22:222-236 (2021)).

[0034] AUF1 has four related protein isoforms identified by their molecular weight(p37AUF1, p40AUF1, p42AUF1, p45AUF1) derived by differential splicing of a single pre-mRNA (Moore et al., “Physiological Networks and Disease Functions of RNA-Binding Protein AUF1,” Wiley Interdiscip. Rev. RNA 5(4):549-564 (2014); Chen & Shyu, “AU-Rich Elements: Characterization and Importance in mRNA Degradation,” Trends Biochem. Sci. 20(11):465-470 (1995); and Kim et al., “Emerging Roles of RNA and RNA-Binding Protein Network in Cancer Cells,” BMB Rep. 42(3): 125-130 (2009), which are hereby incorporated by reference in their entirety). Each of these four isoforms include two centrally-positioned, tandemly arranged RNA recognition motifs (“RRMs”) which mediate RNA binding (DeMaria et al., “Structural Determinants in AUF 1 Required for High Affinity Binding to A+U-rich Elements,” J. Biol. Chem. 21221635-216N3 (1997), which is hereby incorporated by reference in its entirety).

[0035] The term “fragment” or “portion” when used herein with respect to a given polypeptide sequence (e.g., AUF1), refers to a contiguous stretch of amino acids of the given polypeptide’s sequence that is shorter than the given polypeptide’s full-length sequence. A fragment of a polypeptide may be defined by its first position and its final position, in which the first and final positions each correspond to a position in the sequence of the given full-length polypeptide. The sequence position corresponding to the first position is situated N-terminal to the sequence position corresponding to the final position. The sequence of the fragment or portion is the contiguous amino acid sequence or stretch of amino acids in the given polypeptide that begins at the sequence position corresponding to the first position and ends at the sequence position corresponding to the final position. Functional or active fragments are fragments that retain functional characteristics, e.g., of the native sequence or other reference sequence.Typically, active fragments are fragments that retain substantially the same activity as the wildtype protein. A fragment may, for example, contain a functionally important domain, such as a domain that is important for receptor or ligand binding.

[0036] Accordingly, in certain embodiments, functional fragments of AUF1 as described herein include at least one RNA recognition domain (“RRM”) domain. In certain embodiments, functional fragments of AUF1 as described herein include two RRM domains.

[0037] AUF1 or functional fragments thereof as described herein may be derived from a mammalian AUF 1. In one embodiment, the AUF 1 or functional fragment thereof is a human AUF1 or functional fragment thereof. In another embodiment, the AUF1 or functional fragment thereof is a murine AUF1 or a functional fragment thereof. The AUF1 protein according to embodiments described herein may include one or more of the AUF1 isoforms p37AUF1, p40AUF1, p42AUF1, and p45AUF1. The GenBank accession numbers corresponding to the nucleotide and amino acid sequences of each human and mouse isoform is found in Table 1 below, each of which is hereby incorporated by reference in its entirety.Table 1: Summary of GenBank Accession Numbers of AUF1 Sequences

[0038] The sequences referred to in Table 1 are reproduced below.

[0039] The human p37AUF1nucleotide sequence of GenBank Accession No.NM_001003810.2 (SEQ ID NO:9) is as follows:CTTCCGTCGGCCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGCGGCCGCCGCTGGTGCTTATTCTTTTTTAGTGCAGCGGGAGAGAGCGGGAGTGTGCGCCGCGCGAGAGTGGGAGGCGAAGGGGGCAGGCCAGGGAGAGGCGCAGGAGCCTTTGCAGCCACGCGCGCGCCTTCCCTGTCTTGTGTGCTTCGCGAGGTAGAGCGGGCGCGCGGCAGCGGCGGGGATTACTTTGCTGCTAGTTTCGGTTCGCGGCAGCGGCGGGTGTAGTCTCGGCGGCAGCGGCGGAGACACTAGCACTATGTCGGAGGAGCAGTTCGGCGGGGACGGGGCGGCGGCAGCGGCAACGGCGGCGGTAGGCGGCTCGGCGGGCGAGCAGGAGGGAGCCATGGTGGCGGCGACACAGGGGGCAGCGGCGGCGGCGGGAAGCGGAGCCGGGACCGGGGGCGGAACCGCGTCTGGAGGCACCGAAGGGGGCAGCGCCGAGTCGGAGGGGGCGAAGATTGACGCCAGTAAGAACGAGGAGGATGAAGGGAAAATGTTTATAGGAGGCCTTAGC T GGGACAC TACAAAGAAAGAT C T GAAGGAC TAG T T T T CCAAAT T T GGT GAAGT T GTAGAC T GCA CTCTGAAGTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAATCGGAGAGTGTAGATAAGGTCATGGATCAAAAAGAACATAAATTGAATGGGAAGGTGATTGATCCT AAAAGGGCCAAAGCCATGAAAACAAAAGAGCCGGTTAAAAAAATTTTTGTTGGTGGCCTTTCTC CAGATACACCTGAAGAGAAAATAAGGGAGTACTTTGGTGGTTTTGGTGAGGTGGAATCCATAGA GCTCCCCATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGCTTTATTACCTTTAAGGAAGAA GAAC GAG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AA AAGTAGCCATGTCGAAGGAACAATATCAGCAACAGCAACAGTGGGGATCTAGAGGAGGATTTGC AGGAAGAGCTCGTGGAAGAGGTGGTGACCAGCAGAGTGGTTATGGGAAGGTATCCAGGCGAGGT GGTCATCAAAATAGCTACAAACCATACTAAATTATTCCATTTGCAACTTATCCCCAACAGGTGG TGAAGCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACCTGCTAATAGCAG TTCAAACTAAATTTTTTGTATCAAGTCCCTGAATGGAAGTATGACGTTGGGTCCCTCTGAAGTT TAATTCTGAGTTCTCATTAAAAGAAATTTGCTTTCATTGTTTTATTTCTTAATTGCTATGCTTC AGAATCAATTTGTGTTTTATGCCCTTTCCCCCAGTATTGTAGAGCAAGTCTTGTGTTAAAAGCC CAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAATCCTTTTGTATAAA AAT GTAT TGGCTCTTT TAT CAT CAGAATAGGAAAAAT T GT CAT GGAT T CAAGT TAT TAAAAGCA TAAGTTTGGAAGACAGGCTTGCCGAAATTGAGGACATGATTAAAATTGCAGTGAAGTTTGAAAT GTTTTTAGCAAAATCTAATTTTTGCCATAATGTGTCCTCCCTGTCCAAATTGGGAATGACTTAA TGTCAATTTGTTTGTTGGTTGTTTTAATAATACTTCCTTATGTAGCCATTAAGATTTATATGAA TATTTTCCCAAATGCCCAGTTTTTGCTTAATATGTATTGTGCTTTTTAGAACAAATCTGGATAA AT G T G C AAAAG TACCCCTTTG C AC AGAT AG TTAATGTTTTATGCTTCCAT T AAAT AAAAAG GAC TTAAAATCTGTTAATTATAATAGAAATGCGGCTAGTTCAGAGAGATTTTTAGAGCTGTGGTGGA C T T CAT AGAT GAAT T C AAG T G T T GAG G GAG GAT T AAAGAAAT AT AT AC CGTGTTTATGTGTGTG TGCTT

[0040] The human p37AUF1amino acid sequence of GenBank Accession No.NP_001003810.1 (SEQ ID NO: 10) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESE GAKIDASKNEEDEGKMFIGGLSWDTTKKDLKDYFSKFGEWDCTLKLDPI TGRSRGFGFVLFKE SESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPDTPEEKIREYFGGFGEVES I ELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKVAMSKEQYQQQQQWGSRGGF AGRARGRGGDQQSGYGKVSRRGGHQNSYKPY

[0041] The human p40AUF1nucleotide sequence of GenBank Accession No.NM_002138.3 (SEQ ID NO: 13) is as follows:CTTCCGTCGGCCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGCGG CCGCCGCTGGTGCTTATTCTTTTTTAGTGCAGCGGGAGAGAGCGGGAGTGTGCGCCGCGCGAGA GTGGGAGGCGAAGGGGGCAGGCCAGGGAGAGGCGCAGGAGCCTTTGCAGCCACGCGCGCGCCTTCCCTGTCTTGTGTGCTTCGCGAGGTAGAGCGGGCGCGCGGCAGCGGCGGGGATTACTTTGCTGC TAGTTTCGGTTCGCGGCAGCGGCGGGTGTAGTCTCGGCGGCAGCGGCGGAGACACTAGCACTAT GTCGGAGGAGCAGTTCGGCGGGGACGGGGCGGCGGCAGCGGCAACGGCGGCGGTAGGCGGCTCG GCGGGCGAGCAGGAGGGAGCCATGGTGGCGGCGACACAGGGGGCAGCGGCGGCGGCGGGAAGCG GAGCCGGGACCGGGGGCGGAACCGCGTCTGGAGGCACCGAAGGGGGCAGCGCCGAGTCGGAGGG GGCGAAGATTGACGCCAGTAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACAC TCTGAAGCAGCGACGGCACAGCGGGAAGAATGGAAAATGTTTATAGGAGGCCTTAGCTGGGACA C T AC AAAGAAAGAT C T GAAG GAG T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAA GTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAATCGGAGAGT GTAGATAAGGT CAT GGAT CAAAAAGAACATAAAT T GAAT GGGAAGGT GAT T GAT CC TAAAAGGG CCAAAGCCATGAAAACAAAAGAGCCGGTTAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGATAC ACCTGAAGAGAAAATAAGGGAGTACTTTGGTGGTTTTGGTGAGGTGGAATCCATAGAGCTCCCC ATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGCTTTATTACCTTTAAGGAAGAAGAACCAG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG C CATGTCGAAGGAACAATATCAGCAACAGCAACAGTGGGGATCTAGAGGAGGATTTGCAGGAAGA GCTCGTGGAAGAGGTGGTGACCAGCAGAGTGGTTATGGGAAGGTATCCAGGCGAGGTGGTCATC AAAATAGCTACAAACCATACTAAATTATTCCATTTGCAACTTATCCCCAACAGGTGGTGAAGCA GTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACCTGCTAATAGCAGTTCAAAC TAAATTTTTTGTATCAAGTCCCTGAATGGAAGTATGACGTTGGGTCCCTCTGAAGTTTAATTCT GAGTTCTCATTAAAAGAAATTTGCTTTCATTGTTTTATTTCTTAATTGCTATGCTTCAGAATCA ATTTGTGTTTTATGCCCTTTCCCCCAGTATTGTAGAGCAAGTCTTGTGTTAAAAGCCCAGTGTG ACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAATCCTTTTGTATAAAAATGTAT TGGCTCTTT TAT CAT CAGAATAGGAAAAAT T GT CAT GGAT T CAAGT TAT TAAAAGCATAAGT T T GGAAGACAGGCTTGCCGAAATTGAGGACATGATTAAAATTGCAGTGAAGTTTGAAATGTTTTTA GCAAAATCTAATTTTTGCCATAATGTGTCCTCCCTGTCCAAATTGGGAATGACTTAATGTCAAT TTGTTTGTTGGTTGTTTTAATAATACTTCCTTATGTAGCCATTAAGATTTATATGAATATTTTC CCAAATGCCCAGTTTTTGCTTAATATGTATTGTGCTTTTTAGAACAAATCTGGATAAATGTGCA AAAG TACCCCTTTG C AC AGAT AG TTAATGTTTTATGCTTCCAT T AAAT AAAAAG GAC T T AAAAT CTGTTAATTATAATAGAAATGCGGCTAGTTCAGAGAGATTTTTAGAGCTGTGGTGGACTTCATA GAT GAAT T C AAG T G T T GAG G GAG GAT T AAAGAAAT AT AT AC C G T G T T T AT G T G T G T G T G C T T

[0042] The human p40AUF1amino acid sequence of GenBank Accession No.NP_002129.2 (SEQ ID NO: 14) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESE GAKIDASKNEEDEGHSNSSPRHSEAATAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEWDCTL KLDPI TGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPDTPEEKIREYFGGFGEVES IELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKVAMSKEQYQQQQQWGSRGGFAGRARGRGGDQQSGYGKVSRRGGHQNSYKPY

[0043] The human p42AUF1nucleotide sequence of GenBank Accession No.NM_031369.2 (SEQ ID NO: 17) is as follows:CTTCCGTCGGCCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGCGG CCGCCGCTGGTGCTTATTCTTTTTTAGTGCAGCGGGAGAGAGCGGGAGTGTGCGCCGCGCGAGA GTGGGAGGCGAAGGGGGCAGGCCAGGGAGAGGCGCAGGAGCCTTTGCAGCCACGCGCGCGCCTT CCCTGTCTTGTGTGCTTCGCGAGGTAGAGCGGGCGCGCGGCAGCGGCGGGGATTACTTTGCTGC TAGTTTCGGTTCGCGGCAGCGGCGGGTGTAGTCTCGGCGGCAGCGGCGGAGACACTAGCACTAT GTCGGAGGAGCAGTTCGGCGGGGACGGGGCGGCGGCAGCGGCAACGGCGGCGGTAGGCGGCTCG GCGGGCGAGCAGGAGGGAGCCATGGTGGCGGCGACACAGGGGGCAGCGGCGGCGGCGGGAAGCG GAGCCGGGACCGGGGGCGGAACCGCGTCTGGAGGCACCGAAGGGGGCAGCGCCGAGTCGGAGGG GGCGAAGATTGACGCCAGTAAGAACGAGGAGGATGAAGGGAAAATGTTTATAGGAGGCCTTAGC T GGGACAC TACAAAGAAAGAT C T GAAGGAC TAG T T T T CCAAAT T T GGT GAAGT T GTAGAC T GCA CTCTGAAGTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAATC GGAGAGTGTAGATAAGGTCATGGATCAAAAAGAACATAAATTGAATGGGAAGGTGATTGATCCT AAAAGGGCCAAAGCCATGAAAACAAAAGAGCCGGTTAAAAAAATTTTTGTTGGTGGCCTTTCTC CAGATACACCTGAAGAGAAAATAAGGGAGTACTTTGGTGGTTTTGGTGAGGTGGAATCCATAGA GCTCCCCATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGCTTTATTACCTTTAAGGAAGAA GAAC GAG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AA AAGTAGCCATGTCGAAGGAACAATATCAGCAACAGCAACAGTGGGGATCTAGAGGAGGATTTGC AGGAAGAGCTCGTGGAAGAGGTGGTGGCCCCAGTCAAAACTGGAACCAGGGATATAGTAACTAT TGGAATCAAGGCTATGGCAACTATGGATATAACAGCCAAGGTTACGGTGGTTATGGAGGATATG ACTACACTGGTTACAACAACTACTATGGATATGGTGATTATAGCAACCAGCAGAGTGGTTATGG GAAGGTATCCAGGCGAGGTGGTCATCAAAATAGCTACAAACCATACTAAATTATTCCATTTGCA ACTTATCCCCAACAGGTGGTGAAGCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCC T G C C AC C T G C T AAT AG C AG T T C AAAC T AAAT T T T T T G T AT C AAG T C C C T GAAT G GAAG T AT GAC GTTGGGTCCCTCTGAAGTTTAATTCTGAGTTCTCATTAAAAGAAATTTGCTTTCATTGTTTTAT TTCTTAATTGCTATGCTTCAGAATCAATTTGTGTTTTATGCCCTTTCCCCCAGTATTGTAGAGC AAGTCTTGTGTTAAAAGCCCAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTA AT AAAT C C T T T T G T AT AAAAAT G TAT TGGCTCTTT TAT CAT CAGAAT AGGAAAAAT T G T CAT GG AT T CAAGT TAT TAAAAGCATAAGT T T GGAAGACAGGC T T GCCGAAAT T GAGGACAT GAT TAAAA TTGCAGTGAAGTTTGAAATGTTTTTAGCAAAATCTAATTTTTGCCATAATGTGTCCTCCCTGTC CAAATTGGGAATGACTTAATGTCAATTTGTTTGTTGGTTGTTTTAATAATACTTCCTTATGTAG CCATTAAGATTTATATGAATATTTTCCCAAATGCCCAGTTTTTGCTTAATATGTATTGTGCTTTT T AGAAC AAAT C T G GAT AAAT G T G C AAAAG TACCCCTTTG C AC AGAT AG TTAATGTTTTATGCT T C CAT T AAAT AAAAAGGAC T T AAAAT C T G T T AAT T AT AAT AGAAAT GC GGC T AG T T CAGAGAGA T T T T TAGAGC T GT GGT GGAC T T CATAGAT GAAT T CAAGT GT T GAGGGAGGAT TAAAGAAATATA TACCGTGTTTATGTGTGTGTGCTT

[0044] The human p42AUF1amino acid sequence of GenBank Accession No.NP_112737.1 (SEQ ID NO: 18) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESE GAKIDASKNEEDEGKMFIGGLSWDTTKKDLKDYFSKFGEWDCTLKLDPI TGRSRGFGFVLFKE SESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPDTPEEKIREYFGGFGEVES I ELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKVAMSKEQYQQQQQWGSRGGF AGRARGRGGGPSQNWNQGYSNYWNQGYGNYGYNSQGYGGYGGYDYTGYNNYYGYGDYSNQQSGY GKVSRRGGHQNSYKPY

[0045] The human p45AUF1nucleotide sequence of GenBank Accession No.NM_031370.2 (SEQ ID NO:21) is as follows:CTTCCGTCGGCCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGCGG CCGCCGCTGGTGCTTATTCTTTTTTAGTGCAGCGGGAGAGAGCGGGAGTGTGCGCCGCGCGAGA GTGGGAGGCGAAGGGGGCAGGCCAGGGAGAGGCGCAGGAGCCTTTGCAGCCACGCGCGCGCCTT CCCTGTCTTGTGTGCTTCGCGAGGTAGAGCGGGCGCGCGGCAGCGGCGGGGATTACTTTGCTGC TAGTTTCGGTTCGCGGCAGCGGCGGGTGTAGTCTCGGCGGCAGCGGCGGAGACACTAGCACTAT GTCGGAGGAGCAGTTCGGCGGGGACGGGGCGGCGGCAGCGGCAACGGCGGCGGTAGGCGGCTCG GCGGGCGAGCAGGAGGGAGCCATGGTGGCGGCGACACAGGGGGCAGCGGCGGCGGCGGGAAGCG GAGCCGGGACCGGGGGCGGAACCGCGTCTGGAGGCACCGAAGGGGGCAGCGCCGAGTCGGAGGG GGCGAAGATTGACGCCAGTAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACAC TCTGAAGCAGCGACGGCACAGCGGGAAGAATGGAAAATGTTTATAGGAGGCCTTAGCTGGGACA C T AC AAAGAAAGAT C T GAAG GAC T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAA GTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAATCGGAGAGT GTAGATAAGGT CAT GGAT CAAAAAGAACATAAAT T GAAT GGGAAGGT GAT T GAT CC TAAAAGGG CCAAAGCCATGAAAACAAAAGAGCCGGTTAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGATAC ACCTGAAGAGAAAATAAGGGAGTACTTTGGTGGTTTTGGTGAGGTGGAATCCATAGAGCTCCCC ATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGCTTTATTACCTTTAAGGAAGAAGAACCAG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG CCATGTCGAAGGAACAATATCAGCAACAGCAACAGTGGGGATCTAGAGGAGGATTTGCAGGAAGA GCTCGTGGAAGAGGTGGTGGCCCCAGTCAAAACTGGAACCAGGGATATAGTAACTATTGGAATC AAGGCTATGGCAACTATGGATATAACAGCCAAGGTTACGGTGGTTATGGAGGATATGACTACACTGGTTACAACAACTACTATGGATATGGTGATTATAGCAACCAGCAGAGTGGTTATGGGAAGGTA TCCAGGCGAGGTGGTCATCAAAATAGCTACAAACCATACTAAATTATTCCATTTGCAACTTATC CCCAACAGGTGGTGAAGCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACC TGCTAATAGCAGTTCAAACTAAATTTTTTGTATCAAGTCCCTGAATGGAAGTATGACGTTGGGT CCCTCTGAAGTTTAATTCTGAGTTCTCATTAAAAGAAATTTGCTTTCATTGTTTTATTTCTTAA TTGCTATGCTTCAGAATCAATTTGTGTTTTATGCCCTTTCCCCCAGTATTGTAGAGCAAGTCTT GTGTTAAAAGCCCAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAATC C T T T T GTATAAAAAT GTAT TGGCTCTTT TAT CAT CAGAATAGGAAAAAT T GT CAT GGAT T CAAG T TAT TAAAAGCATAAGT T T GGAAGACAGGC T T GCCGAAAT T GAGGACAT GAT TAAAAT T GCAGT GAAGTTTGAAATGTTTTTAGCAAAATCTAATTTTTGCCATAATGTGTCCTCCCTGTCCAAATTG GGAATGACTTAATGTCAATTTGTTTGTTGGTTGTTTTAATAATACTTCCTTATGTAGCCATTAA GATTTATATGAATATTTTCCCAAATGCCCAGTTTTTGCTTAATATGTATTGTGCTTTTTAGAAC AAAT C T G GAT AAAT G T G C AAAAG T AC C C C T T T G C AC AGAT AG T T AAT G T T T T AT G C T T C C AT T A AATAAAAAGGAC T TAAAAT C T GT TAAT TATAATAGAAAT GCGGC TAGT T CAGAGAGAT T T T TAG AGCTGTGGTGGACTTCATAGATGAATTCAAGTGTTGAGGGAGGATTAAAGAAATATATACCGTG TTTATGTGTGTGTGCTT

[0046] The human p45AUF1amino acid sequence of GenBank Accession No.NP_112738.1 (SEQ ID NO:22) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAATQGAAAAAGSGAGTGGGTASGGTEGGSAESE GAKIDASKNEEDEGHSNSSPRHSEAATAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEWDCTL KLDPI TGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPD TPEEKIREYFGGFGEVES IELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKV AMSKEQYQQQQQWGSRGGFAGRARGRGGGPSQNWNQGYSNYWNQGYGNYGYNSQGYGGYGGYDY TGYNNYYGYGDYSNQQSGYGKVSRRGGHQNSYKPY

[0047] The mouse p37AUF1nucleotide sequence of GenBank Accession No.NM_001077267.2 (SEQ ID NO: 11) is as follows:CCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGTGGCCGCCGCTGC TACTTCATTCTTTTTTTTTTCAGTGCAGCCGGGGAGAGCGAGAGAGCGCGCTGCGCGAGAGTGG GAGGCGAGGGGGGCAGGCCGGGGAGAGGCGCAGGAGCCCTTGCAGCCACGCGCGCGCCTTGTCT AGGGTGCCTCGCGAGGTAGAGCGGGCATCGCGCGGCGGCGGCGGGGATTACTTTGCTGCTAGTT TCGGTTCGCGGCGGCGGCGGCGTCGGCGGGTGTCGTCTTCGGCGGCGGCAGTAGCACTATGTCG GAGGAGCAGTTCGGAGGGGACGGGGCGGCGGCGGCGGCAACGGCGGCGGTAGGCGGCTCGGCGG GCGAGCAGGAGGGAGCCATGGTGGCGGCGGCGGCGCAGGGGCCGGCGGCGGCGGCGGGAAGCGG GAGCGGCGGCGGCGGCTCTGCGGCCGGAGGCACCGAAGGAGGCAGCGCCGAGGCAGAGGGAGCCAAGATCGACGCCAGTAAGAACGAGGAGGATGAAGGGAAAATGTTTATAGGAGGCCTTAGCTGGG AC AC C AC AAAGAAAGAT C T GAAG GAC T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAAGTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAGTCGGAG AG T G T AGAT AAG G T C AT G GAT C AGAAAGAAC AT AAAT T GAAT G G GAAAG T C AT T GAT C C T AAAA GGGCCAAAGCCATGAAAACAAAAGAGCCTGTCAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGA CACACCTGAAGAAAAAATAAGAGAGTACTTTGGTGGTTTTGGTGAGGTTGAATCCATAGAGCTC CCTATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGTTTTATTACCTTTAAGGAAGAGGAGC C AG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AGCCATGTCAAAGGAACAGTATCAGCAGCAGCAGCAGTGGGGATCTAGAGGAGGGTTTGCAGGC AGAGCTCGCGGAAGAGGTGGAGATCAGCAGAGTGGTTATGGGAAAGTATCCAGGCGAGGTGGAC AT C AAAAT AG C T AC AAAC CAT AC T AAAT TATTCCATTTG C AAC T T AT C C C C AAC AG G T G G T GAA GCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACCTGCTAATAGCAGTTCA AACTAAATTTTTTCTATCAAGTTCCTGAATGGAAGTATGACGTTGGGTCCCTCTGAAGTTTAAT TCTGAGTTCTCATTAAAAGAATTTGCTTTCATTGTTTTATTTCTTAATTGCTATGCTTCAGTAT CAATTTGTGTTTTATGCCCCCCCTCCCCCCCAGTATTGTAGAGCAAGTCTTGTGTTAAAAAAAG CCCAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAATCCTTTTGTATA AAAAT G TAT TGGCTCTTT TAT CAT CAGAAT AGGAGGAAG T GAAAT AC T ACAAAT GTTTGTCTTG GAT T CAAGT CAC TAGAAGCATAAAT T T GAGGGGATAAAAACAACGGTAAAC T T T GT C T GAAAGA GGGCATGGTTAAAAATGTAGTGAATTTTAAATGTTTTTAGCAAAATTTGATTTTGCCCAAGAAT CCCTGTCTGAATTGGAAATGACTTAATGTAGTCAATGTGCTTGTTGGTTGTCTTAATATTACTT CTGTAGCCATTAAGTTTTATGAGTAACTTCCCAAATACCCACGTTTTTCTTTATATGTATTGTG C T T T T TAAAAACAAAT C T GGAAAAAT GGGCAAGAACAT T T GCAGACAAT T GT T T T TAAGC T T CC AT TAAATAAAAAAAAT GT GGAC T TAAGGAAAT C TAT TAAT T TAAATAGAAC T GCAGC TAGT T TA GAGAGTATTTTTTTCTTAAAGCTTTGGTGTAATTAGGGAAGATTTTAAAAAATGCATAGTGTTT ATTTGTATGTGTGCTCTTTTTTTAAGTCAATTTTTGGGGGGTTGGTCTGTTAACTGAGTCTAGG ATTTAAAGGTAAGATGTTCCTAGAAATCTTGTCATCCCAAAGGGGCGGGCGCTAAGGTGAAACT TCAGGGTTCAGTCAGGGTCACTGCTTTATGTGTGAAATCACTCAAATTGGTAAGTCTCTTATGT T AGCAT T CAGGACAT T GAT T T CAAC T T GGAT GGACAAT T TAT AG T TAG TAG T GAAT TGTGTGTT AATGTGTTCAGTCCTGGTAAGTTTTCAGTTTGATCAGTTAGTTGGAAGCAGACTTGAAGAGCTG TTAGTCACGTGAGCCATGGGTGCAGTCGATCTGTGGTCAGATGCCTGAGTCTGTGATAGTGAAT T G T G T C T AAAGAC AT T T T AAT GAT AAAAG T GAG T G C T G T AAAG T T GAAAG T T C AT GAGAGAC AT ACAAT GAGGGC T GCAGCCCAT T T T TAAAAACAT TATAATACAAAAGTAT GCACAT T T GT T TACA TATCCCTGCCTTTGTATTACAGTGGCAGGTTTGTGTACTTAAACTGGGAAAGCCTCAGATCTAT GATTACCTGGCCTAT C AT AGAAAG T G T C T AAAT AAAT CAC T C T G T GAAT T GAAT AC AT TAG TAT TAGCTAGCATACTTCATTATGCCTGTTTTCCATAAATACCACACCAAAAACTTGCTTGGGGCAGTTTGAGCCTAGTTCATGAGCTGCTATCAGATTGGTCTTGATCCTATATAATAGGCCAAATGTCT GTAAACAGCTGTGCTGGTGGAATGTAGAAAGTCACTGCACTCAGATTCAACTTCCTGATTGGAA G T C AT GAG AG T G T GAT T AAAC AT T T T C AC AAAGAAT AG T AGAT AAAT AAC TTGGTTTTTAATGT TAACTTTGTTTCCATTAAGTCACATTTAAAAACTTATCCTCACGCCTACCTGAGTTAATTATCT GTTGACCTAGATATCTTTCTGGCCACTCACTGACTTATTTCTTGAACTTTTGCCATTTGCATAA ATCTTGTCAGCTTTGTTCTTGATTATGCATTGTCCAGGCTGAGCTAGTTGTCTTTCCAGGAATC CCTTTGTCTCTGAATTAGGTCCTTTGTTTCCTAAATCATCCTGCTTGTTTGGCACAAGTCTTCC CAGGCCAGTGAGACCTCCGTGTCCTCTCAGCACCATAGGGGTAGGTAACCCTGGTTAGGCTGGA CAGGGGTTTGCTGAGGGAGTTTGTTCATTTGAATCTAGGTCTTACATGACGTCTTTCAAATAGG GTTTTTACCTTGACACTAAACTGTCCAGTCTAAGCAGTTCTGCAAAATGTGAGGGAATTATGAA CTTCTTCCTGCAGTGGGTTTTTATGGTTTTGGTTTGTTTTTTGTTGTTTTGGTTCTTTGTTGAG C C C T G GAG AAAAAC T T C C C T AG T T C T G G T T T C T AC AAT T T AAAT T AAAAAC AGAAT T C AT C T T A GAATTTTTCACCCTCTTCCCCAACTATTCTAATCAATCTTAAGTATGCCCTTCATCTTTTTTCC TTCCTAAGGCTTTTACTGATAGTGTAATTCCGTACTCTTCAACCCTGGGAAGGCTGAAGTGGAT TCTTGAGCTCATTTCAAGGCTGACCTGGGTGTTGGCAAGAACCCAGCTTAGAACAAACACATGC AAGGCCATCTTACCTTACATCCTGTTGCTTGGACTTCTTCCTGCTCAAAGTTTTTAGTGGATGC T AAG TGATCTTTGCTTC C AC T GAG GAG T G GAAC AC T T T AGAAT GAAC C T C T AGAT AGAT AT T T T TATTGTCTGGTGAGGGTTACTGGAGTTTCCCACCCTGCCTGAAGGGTGAATCTGGCTTACAGTG TTCTCATCTCAAAGGGAAGAAGGCAGATGGCTGTGTCCAGAGAGAGCCATCACAGTTTGCTTCA GAGACACTAGAATGGGCTGGAAGATCTAGTGGTCTTAATCAGACTTGAAACCTGGCCTTTCTTC ATTACCCATATGTCTACCAGTACTTGGGCTAACACTTAAGCCATTAGGGCCTTTGTAGGGGTGT T T T GAGAC CCCCTCCATGC T AAC AAAT AT AC AG G T T T C T T AAC AT T T G C T CAT AAAC T T G T AAA GCTTACTTTCTCTTAATC C AC C C C AC AT T T AAC AAG CCCTGGTACT T AGAAT T T C AGAAGAG TA ATGGCAGGTAGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGAGAGAGAG AGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAG T T T G T G GAAAAT C AG G T AATGACAGCTCATCCTTTTAGAATTGTACTTCAGAATAGAAACATTTGGTGGGCTGTTAGGTAG CTTTGATTACTTGTGGGTAGACCTGCTAGTATTGCCAGTCCTCAAGCAATGAGCTTTCTGTATC TTGTTTACTAGATATATACTACCAGGTGAGTCATTTCCTGGGGTTCTGTTTTCTTTTAAAATCT TTCCCTAAACTTAATATGTATTAAAAAGTCTGGCTTTTCAGTCCATTCTTTGTGCACTGGGATG GCAATTGCTTCATTATATGACAATTGCTGTTCCCAAGTCAGAATTCAGTGTGCTGATTTGACAT CAGTTCGTCCCGAATAAGTTCCTGTTACCAGGATTTACATTCAGCACATTAGAAACTTGTTGGT GTGCTTTTATTCTTGGAGCATTTTCCTTAGACTACCTTCCACTTTGAGTGCTCTGTTTAGGATG TTGAGGTGTTAGGATTCTTGACAGCCAGAAAGACTGAACCCACTATCTGGGCACAGTGTTCGTG TTGCTCTATAAATGTATGCTTTTTTTGATTTGGGGTTGTTTTACCTACATTGTCAAACTAGATC CATGCTTAACAGTGATAATGAAGGCTTTTTGTTTGTTTTGTTTGTGGGTCCTCCCCCCCCCCCCAAGACAGGGTTTCTCTGTAGGCTGTCCTAGAACTTGTTCTTTTTTAACCAAAATTTGGCAAGGC T GAAAAT G GAAT C C T AT AAT C AAT G C T G G C C AC AT T AAAG T T AAT AG T T GAGAAG T C T T G T C T G AATTTCCTTGGGCAAAAAGATTCTAGCCAGTTCAATACCCTGTTGTGCAAATTCAATTTGCTGT TATAATTTGCTCTCAGTTATCAGTTGGAAGGAGGTTAATTCTAATGTACTTGGAAGAGGCCTGT AGACCATCTATAACTGCATCAGTTGTACAGCGTTGTTGCCTGGGATTCTCTAGTTCACATAAAC TCCCAAGTCTTAGCCGTGGTGATGGCTACAGTGTGGAAGATGGTGAGCATTCTAGTGAGTATCG CGATGACGGCAGTAAAGAGCAGCAGGCAGCCGTGGCTGGGCTCACTGACCGTGGCTGTAAGTTA CGGAGGCAGCACACACTTCTGTACACACCTCTCATCAGTTACCGGAGTCATTGCATTGCGGACT AACTGGCTGACTCAAGTTGTCTTGCTACTGAAGTCTTGAGTTGGTCTCATGCATTTACCCTGTT GACTTGAGCACCTTAAAGTCGAAAGGATGTCTGGTTGTGGCTTTATTGTAAACAGCCTTAGGTA AAGAGGGGAGTATATCGGTTAGGAAGGTGAAAAATGATACTTCCAAGTTCAGTGGGAAACCCTG GGTTTATCCCCCAGCTTAAGAAAGAATGCCTAACAATGTTTCAGAATTAGATTCTGTGGAAGGT GAGGGTGTTAGAACAGTCCAAATTTGTTATTGTAGACTTGCAGTGGGAGGAATTTTTAAATATA CAGATCAGTCGACACTCATTAACTTCACTGATAAAGGTGGAAACGGATGTGGCAACACTTCTAA GTTCATTTGTATATGTTTGTAATTTGATTGGTTGTATTCTGTTGCACTCTAGAATTTGAAGGCA AGG T T AC C T C T GC T T T T T AAT T T T T T T T T T T T T AAAGAAAGAAAAAACAC T GAAAGAAAC T T GA AAAGAT C T GT TAAT GC TAATACC T GAAT GT GGCAT T TAACAT GT CAT GGAAAC T GC T T T GAATA AATACTTGAGAAAAGGAATGAAATAATTGCCGTTTTTGTTGTTGAGTGAATGGGTGTGGTTTAA T GAG CGTAATCATTTT T AT AAAAC AG C T G T GAGAC T GAAG T G GAAT C C T T AT T AAAT G T G GAAA ATGGCCTTT GAGGAT T ACAG T AGAGAT T CAAC T AAGAGAG T AAAT AAAGC T T GAAAC TAAT T C G T T GTAAAT T GC T T C TACAAT CAT T GC T C TATATAGCAT GC TAT T GCCAAT CAGT T T TAT GTAT T AAGACCTATCAGCATGTCTTTTTTAGGTTGACCTCATTTTAAATTATAAGATGCTCTCTGTACC GTTTTAACATTTCCAGGATTTATTCTTTCTAGGCAAATTCCACTGGACTGTTTCCATTGTAGAA GCTTCCTTATAGATTCTTCAAATGAAGCTTACAGTGTGCTTTCTTGGGGTTTTGATTTGCACTA AATTTTATTTTCTGAAAGATCACTTATGTTTATAATGTAGTGCTTTGTCTTAACAATTAAACTT TCCAGCACTCATGCA

[0048] The mouse p37AUF1amino acid sequence of GenBank Accession No.NP_001070735.1 (SEQ ID NO: 12) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAAAAQGPAAAAGSGSGGGGSAAGGTEGGSAEAE GAKIDASKNEEDEGKMFIGGLSWDTTKKDLKDYFSKFGEWDCTLKLDPI TGRSRGFGFVLFKE SESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPDTPEEKIREYFGGFGEVES I ELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKVAMSKEQYQQQQQWGSRGGF AGRARGRGGDQQSGYGKVSRRGGHQNSYKPY

[0049] The mouse p40AUF1nucleotide sequence of GenBank Accession No.NM_007516.3 (SEQ ID NO:15) is as follows:CCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGTGGCCGCCGCTGC TACTTCATTCTTTTTTTTTTCAGTGCAGCCGGGGAGAGCGAGAGAGCGCGCTGCGCGAGAGTGG GAGGCGAGGGGGGCAGGCCGGGGAGAGGCGCAGGAGCCCTTGCAGCCACGCGCGCGCCTTGTCT AGGGTGCCTCGCGAGGTAGAGCGGGCATCGCGCGGCGGCGGCGGGGATTACTTTGCTGCTAGTT TCGGTTCGCGGCGGCGGCGGCGTCGGCGGGTGTCGTCTTCGGCGGCGGCAGTAGCACTATGTCG GAGGAGCAGTTCGGAGGGGACGGGGCGGCGGCGGCGGCAACGGCGGCGGTAGGCGGCTCGGCGG GCGAGCAGGAGGGAGCCATGGTGGCGGCGGCGGCGCAGGGGCCGGCGGCGGCGGCGGGAAGCGG GAGCGGCGGCGGCGGCTCTGCGGCCGGAGGCACCGAAGGAGGCAGCGCCGAGGCAGAGGGAGCC AAGATCGACGCCAGTAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACACACTG AAGCAGCGGCGGCACAGCGGGAAGAATGGAAAATGTTTATAGGAGGCCTTAGCTGGGACACCAC AAAGAAAGAT C T GAAG GAG T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAAG T T A GATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAGTCGGAGAGTGTAG ATAAGGT CAT GGAT CAGAAAGAACATAAAT T GAAT GGGAAAGT CAT T GAT CC TAAAAGGGCCAA AGCCATGAAAACAAAAGAGCCTGTCAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGACACACCT GAAGAAAAAATAAGAGAGTACTTTGGTGGTTTTGGTGAGGTTGAATCCATAGAGCTCCCTATGG ACAACAAGACCAATAAGAGGCGTGGGTTCTGTTTTATTACCTTTAAGGAAGAGGAGCCAGTGAA GAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG C C AT G TCAAAGGAACAGTATCAGCAGCAGCAGCAGTGGGGATCTAGAGGAGGGTTTGCAGGCAGAGCTC GCGGAAGAGGTGGAGATCAGCAGAGTGGTTATGGGAAAGTATCCAGGCGAGGTGGACATCAAAA TAGCTACAAACCATACTAAATTATTCCATTTGCAACTTATCCCCAACAGGTGGTGAAGCAGTAT TTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACCTGCTAATAGCAGTTCAAACTAAA TTTTTTCTATCAAGTTCCTGAATGGAAGTATGACGTTGGGTCCCTCTGAAGTTTAATTCTGAGT TCTCATTAAAAGAATTTGCTTTCATTGTTTTATTTCTTAATTGCTATGCTTCAGTATCAATTTG TGTTTTATGCCCCCCCTCCCCCCCAGTATTGTAGAGCAAGTCTTGTGTTAAAAAAAGCCCAGTG TGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAATCCTTTTGTATAAAAATGT ATTGGCTCTTT TAT CAT C AGAAT AG GAG GAAG T GAAAT AC TACAAAT GT T T GT C T T GGAT T CAA GT CAC TAGAAGCATAAAT T T GAGGGGATAAAAACAACGGTAAAC T T T GT C T GAAAGAGGGCAT G GTTAAAAATGTAGTGAATTTTAAATGTTTTTAGCAAAATTTGATTTTGCCCAAGAATCCCTGTC TGAATTGGAAATGACTTAATGTAGTCAATGTGCTTGTTGGTTGTCTTAATATTACTTCTGTAGC CATTAAGTTTTATGAGTAACTTCCCAAATACCCACGTTTTTCTTTATATGTATTGTGCTTTTTA AAAACAAAT C T GGAAAAAT GGGCAAGAACAT T T GCAGACAAT T GT T T T TAAGC T T CCAT TAAAT AAAAAAAAT G T GGAC T T AAGGAAAT C TAT T AAT T TAAAT AGAAC T GCAGC T AG T T T AGAGAG TA TTTTTTTCTTAAAGCTTTGGTGTAATTAGGGAAGATTTTAAAAAATGCATAGTGTTTATTTGTATGTGTGCTCTTTTTTTAAGTCAATTTTTGGGGGGTTGGTCTGTTAACTGAGTCTAGGATTTAAA GGTAAGATGTTCCTAGAAATCTTGTCATCCCAAAGGGGCGGGCGCTAAGGTGAAACTTCAGGGT TCAGTCAGGGTCACTGCTTTATGTGTGAAATCACTCAAATTGGTAAGTCTCTTATGTTAGCATT GAG GAG AT T GAT T T C AAC T T G GAT G GAG AAT T TAT AG T TAG TAG T GAAT TGTGTGTTAATGTGT TCAGTCCTGGTAAGTTTTCAGTTTGATCAGTTAGTTGGAAGCAGACTTGAAGAGCTGTTAGTCA CGTGAGCCATGGGTGCAGTCGATCTGTGGTCAGATGCCTGAGTCTGTGATAGTGAATTGTGTCT AAAGAC AT T T T AAT GAT AAAAG T GAG T G C T G T AAAG T T GAAAG T T C AT GAGAGAC AT AC AAT GA GGGCTGCAGCCCATTTTTAAAAACATTATAATACAAAAGTATGCACATTTGTTTACATATCCCT GCCTTTGTATTACAGTGGCAGGTTTGTGTACTTAAACTGGGAAAGCCTCAGATCTATGATTACC TGGCCTAT C AT AGAAAG T G T C T AAAT AAAT GAG T C T G T GAAT T GAAT AC AT TAG TAT TAG C TAG CATACTTCATTATGCCTGTTTTCCATAAATACCACACCAAAAACTTGCTTGGGGCAGTTTGAGC CTAGTTCATGAGCTGCTATCAGATTGGTCTTGATCCTATATAATAGGCCAAATGTCTGTAAACA GCTGTGCTGGTGGAATGTAGAAAGTCACTGCACTCAGATTCAACTTCCTGATTGGAAGTCATCA C AG T G T GAT T AAAC AT T T T C AC AAAGAAT AG TAGAT AAAT AAC TTGGTTTTTAATGT T AAC T T T GTTTCCATTAAGTCACATTTAAAAACTTATCCTCACGCCTACCTGAGTTAATTATCTGTTGACC TAGATATCTTTCTGGCCACTCACTGACTTATTTCTTGAACTTTTGCCATTTGCATAAATCTTGT CAGCTTTGTTCTTGATTATGCATTGTCCAGGCTGAGCTAGTTGTCTTTCCAGGAATCCCTTTGT CTCTGAATTAGGTCCTTTGTTTCCTAAATCATCCTGCTTGTTTGGCACAAGTCTTCCCAGGCCA GTGAGACCTCCGTGTCCTCTCAGCACCATAGGGGTAGGTAACCCTGGTTAGGCTGGACAGGGGT TTGCTGAGGGAGTTTGTTCATTTGAATCTAGGTCTTACATGACGTCTTTCAAATAGGGTTTTTA CCTTGACACTAAACTGTCCAGTCTAAGCAGTTCTGCAAAATGTGAGGGAATTATGAACTTCTTC CTGCAGTGGGTTTTTATGGTTTTGGTTTGTTTTTTGTTGTTTTGGTTCTTTGTTGAGCCCTGGA C AAAAAC T T C C C TAG T T C T G G T T T C TAG AAT T T AAAT TAAAAACAGAAT T CAT C T TAGAAT T T T TCACCCTCTTCCCCAACTATTCTAATCAATCTTAAGTATGCCCTTCATCTTTTTTCCTTCCTAA GGCTTTTACTGATAGTGTAATTCCGTACTCTTCAACCCTGGGAAGGCTGAAGTGGATTCTTGAG CTCATTTCAAGGCTGACCTGGGTGTTGGCAAGAACCCAGCTTAGAACAAACACATGCAAGGCCA TCTTACCTTACATCCTGTTGCTTGGACTTCTTCCTGCTCAAAGTTTTTAGTGGATGCTAAGTGA TCTTTGCTTCCACTGAGGAGTGGAACACTTTAGAATGAACCTCTAGATAGATATTTTTATTGTC TGGTGAGGGTTACTGGAGTTTCCCACCCTGCCTGAAGGGTGAATCTGGCTTACAGTGTTCTCAT CTCAAAGGGAAGAAGGCAGATGGCTGTGTCCAGAGAGAGCCATCACAGTTTGCTTCAGAGACAC TAGAATGGGCTGGAAGATCTAGTGGTCTTAATCAGACTTGAAACCTGGCCTTTCTTCATTACCC ATATGTCTACCAGTACTTGGGCTAACACTTAAGCCATTAGGGCCTTTGTAGGGGTGTTTTGAGA CCCCCTCCATGC T AAC AAAT AT AC AG G T T T C T T AAC AT T T G C T CAT AAAC T T G T AAAG C T T AC T TTCTCTTAATC C AC C C C AC AT T T AAC AAG CCCTGGTACT TAGAAT T T C AGAAGAG T AAT G G C AG GTAGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAG T T T G T G GAAAAT GAG G T AAT GAGA GCTCATCCTTTTAGAATTGTACTTCAGAATAGAAACATTTGGTGGGCTGTTAGGTAGCTTTGAT TACTTGTGGGTAGACCTGCTAGTATTGCCAGTCCTCAAGCAATGAGCTTTCTGTATCTTGTTTA CTAGATATATACTACCAGGTGAGTCATTTCCTGGGGTTCTGTTTTCTTTTAAAATCTTTCCCTA AACTTAATATGTATTAAAAAGTCTGGCTTTTCAGTCCATTCTTTGTGCACTGGGATGGCAATTG CTTCATTATATGACAATTGCTGTTCCCAAGTCAGAATTCAGTGTGCTGATTTGACATCAGTTCG TCCCGAATAAGTTCCTGTTACCAGGATTTACATTCAGCACATTAGAAACTTGTTGGTGTGCTTT TATTCTTGGAGCATTTTCCTTAGACTACCTTCCACTTTGAGTGCTCTGTTTAGGATGTTGAGGT GTTAGGATTCTTGACAGCCAGAAAGACTGAACCCACTATCTGGGCACAGTGTTCGTGTTGCTCT ATAAATGTATGCTTTTTTTGATTTGGGGTTGTTTTACCTACATTGTCAAACTAGATCCATGCTT AACAGTGATAATGAAGGCTTTTTGTTTGTTTTGTTTGTGGGTCCTCCCCCCCCCCCCAAGACAG GGTTTCTCTGTAGGCTGTCCTAGAACTTGTTCTTTTTTAACCAAAATTTGGCAAGGCTGAAAAT G GAAT C C T AT AAT C AAT G C T G G C C AC AT T AAAG T T AAT AG T T GAGAAG T C T T G T C T GAAT T T C C TTGGGCAAAAAGATTCTAGCCAGTTCAATACCCTGTTGTGCAAATTCAATTTGCTGTTATAATT TGCTCTCAGTTATCAGTTGGAAGGAGGTTAATTCTAATGTACTTGGAAGAGGCCTGTAGACCAT CTATAACTGCATCAGTTGTACAGCGTTGTTGCCTGGGATTCTCTAGTTCACATAAACTCCCAAG TCTTAGCCGTGGTGATGGCTACAGTGTGGAAGATGGTGAGCATTCTAGTGAGTATCGCGATGAC GGCAGTAAAGAGCAGCAGGCAGCCGTGGCTGGGCTCACTGACCGTGGCTGTAAGTTACGGAGGC AGCACACACTTCTGTACACACCTCTCATCAGTTACCGGAGTCATTGCATTGCGGACTAACTGGC TGACTCAAGTTGTCTTGCTACTGAAGTCTTGAGTTGGTCTCATGCATTTACCCTGTTGACTTGA GCACCTTAAAGTCGAAAGGATGTCTGGTTGTGGCTTTATTGTAAACAGCCTTAGGTAAAGAGGG GAGTATATCGGTTAGGAAGGTGAAAAATGATACTTCCAAGTTCAGTGGGAAACCCTGGGTTTAT CCCCCAGCTTAAGAAAGAATGCCTAACAATGTTTCAGAATTAGATTCTGTGGAAGGTGAGGGTG T T AGAAC AG T C C AAAT TTGTTATTG T AGAC T T G GAG T G G GAG GAAT T T T T AAAT AT AC AGAT C A GTCGACACTCATTAACTTCACTGATAAAGGTGGAAACGGATGTGGCAACACTTCTAAGTTCATT TGTATATGTTTGTAATTTGATTGGTTGTATTCTGTTGCACTCTAGAATTTGAAGGCAAGGTTAC C T C T GC T T T T TAAT T T T T T T T T T T T TAAAGAAAGAAAAAACAC T GAAAGAAAC T T CAAAAGAT C T GT TAAT GC TAATACC T GAAT GT GGCAT T TAACAT GT CAT GGAAAC T GC T T T GAAT AAAT AC T T GAGAAAAGGAATGAAATAATTGCCGTTTTTGTTGTTGAGTGAATGGGTGTGGTTTAATGAGCGT AATCATTTTTATAAAACAGCTGTGAGACTGAAGTGGAATCCTTATTAAATGTGGAAAATGGCCT T T GAG GAT TAG AG T AGAGAT T C AAC T AAGAGAG T AAAT AAAG C T T GAAAC TAATTCGTTG T AAA T T GC T T C TACAAT CAT T GC T C TATATAGCAT GC TAT T GCCAAT CAGT T T TAT GTAT TAAGACC T ATCAGCATGTCTTTTTTAGGTTGACCTCATTTTAAATTATAAGATGCTCTCTGTACCGTTTTAA CATTTCCAGGATTTATTCTTTCTAGGCAAATTCCACTGGACTGTTTCCATTGTAGAAGCTTCCT TATAGATTCTTCAAATGAAGCTTACAGTGTGCTTTCTTGGGGTTTTGATTTGCACTAAATTTTATTTTCTGAAAGATCACTTATGTTTATAATGTAGTGCTTTGTCTTAACAATTAAACTTTCCAGCA CT CAT GCA

[0050] The mouse p40AUF1amino acid sequence of GenBank Accession No.NP-031542.2 (SEQ ID NO: 16) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAAAAQGPAAAAGSGSGGGGSAAGGTEGGSAEAE GAKIDASKNEEDEGHSNSSPRHTEAAAAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEWDCTL KLDPI TGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPD TPEEKIREYFGGFGEVES IELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKV AMSKEQYQQQQQWGSRGGFAGRARGRGGDQQSGYGKVSRRGGHQNSYKPY

[0051] The mouse p42AUF1nucleotide sequence of GenBank Accession No.NM_001077266.2 (SEQ ID NO: 19) is as follows:CCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGTGGCCGCCGCTGC TACTTCATTCTTTTTTTTTTCAGTGCAGCCGGGGAGAGCGAGAGAGCGCGCTGCGCGAGAGTGG GAGGCGAGGGGGGCAGGCCGGGGAGAGGCGCAGGAGCCCTTGCAGCCACGCGCGCGCCTTGTCT AGGGTGCCTCGCGAGGTAGAGCGGGCATCGCGCGGCGGCGGCGGGGATTACTTTGCTGCTAGTT TCGGTTCGCGGCGGCGGCGGCGTCGGCGGGTGTCGTCTTCGGCGGCGGCAGTAGCACTATGTCG GAGGAGCAGTTCGGAGGGGACGGGGCGGCGGCGGCGGCAACGGCGGCGGTAGGCGGCTCGGCGG GCGAGCAGGAGGGAGCCATGGTGGCGGCGGCGGCGCAGGGGCCGGCGGCGGCGGCGGGAAGCGG GAGCGGCGGCGGCGGCTCTGCGGCCGGAGGCACCGAAGGAGGCAGCGCCGAGGCAGAGGGAGCC AAGATCGACGCCAGTAAGAACGAGGAGGATGAAGGGAAAATGTTTATAGGAGGCCTTAGCTGGG AC AC C AC AAAGAAAGAT C T GAAG GAC T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAAGTTAGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAGTCGGAG AG T G T AGAT AAG G T C AT G GAT C AGAAAGAAC AT AAAT T GAAT G G GAAAG T C AT T GAT C C T AAAA GGGCCAAAGCCATGAAAACAAAAGAGCCTGTCAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGA CACACCTGAAGAAAAAATAAGAGAGTACTTTGGTGGTTTTGGTGAGGTTGAATCCATAGAGCTC CCTATGGACAACAAGACCAATAAGAGGCGTGGGTTCTGTTTTATTACCTTTAAGGAAGAGGAGC C AG T GAAGAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AGCCATGTCAAAGGAACAGTATCAGCAGCAGCAGCAGTGGGGATCTAGAGGAGGGTTTGCAGGC AGAGCTCGCGGAAGAGGTGGAGGCCCCAGTCAAAACTGGAACCAGGGATATAGTAACTATTGGA ATCAAGGCTATGGCAACTATGGATATAACAGCCAAGGTTACGGAGGTTATGGAGGATATGACTA CACTGGTTACAACAACTACTATGGATATGGTGATTATAGCAATCAGCAGAGTGGTTATGGGAAA GTATCCAGGCGAGGTGGACATCAAAATAGCTACAAACCATACTAAATTATTCCATTTGCAACTT ATCCCCAACAGGTGGTGAAGCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCC ACCTGCTAATAGCAGTTCAAACTAAATTTTTTCTATCAAGTTCCTGAATGGAAGTATGACGTTGGGTCCCTCTGAAGTTTAATTCTGAGTTCTCATTAAAAGAATTTGCTTTCATTGTTTTATTTCTT AATTGCTATGCTTCAGTATCAATTTGTGTTTTATGCCCCCCCTCCCCCCCAGTATTGTAGAGCA AGTCTTGTGTTAAAAAAAGCCCAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTT T AAT AAAT C C T T T T G T AT AAAAAT G TAT TGGCTCTTT TAT CAT CAGAAT AGGAGGAAG T GAAAT AC TACAAAT GT T T GT C T T GGAT T CAAGT CAC TAGAAGCATAAAT T T GAGGGGATAAAAACAACG GTAAACTTTGTCTGAAAGAGGGCATGGTTAAAAATGTAGTGAATTTTAAATGTTTTTAGCAAAA TTTGATTTTGCCCAAGAATCCCTGTCTGAATTGGAAATGACTTAATGTAGTCAATGTGCTTGTT GGTTGTCTTAATATTACTTCTGTAGCCATTAAGTTTTATGAGTAACTTCCCAAATACCCACGTT T T T C T T TATAT GTAT T GT GC T T T T TAAAAACAAAT C T GGAAAAAT GGGCAAGAACAT T T GCAGA CAAT T G T T T T T AAGC T T C CAT T AAAT AAAAAAAAT G T GGAC T T AAGGAAAT C TAT T AAT T T AAA TAGAACTGCAGCTAGTTTAGAGAGTATTTTTTTCTTAAAGCTTTGGTGTAATTAGGGAAGATTT TAAAAAATGCATAGTGTTTATTTGTATGTGTGCTCTTTTTTTAAGTCAATTTTTGGGGGGTTGG TCTGTTAACTGAGTCTAGGATTTAAAGGTAAGATGTTCCTAGAAATCTTGTCATCCCAAAGGGG CGGGCGCTAAGGTGAAACTTCAGGGTTCAGTCAGGGTCACTGCTTTATGTGTGAAATCACTCAA AT T GGTAAGT C T C T TAT GT TAGCAT T CAGGACAT T GAT T T CAAC T T GGAT GGACAAT T TATAGT TACTACTGAATTGTGTGTTAATGTGTTCAGTCCTGGTAAGTTTTCAGTTTGATCAGTTAGTTGG AAGCAGACTTGAAGAGCTGTTAGTCACGTGAGCCATGGGTGCAGTCGATCTGTGGTCAGATGCC T GAG T C T G T GAT AG T GAAT T G T G T C T AAAGAC AT T T T AAT GAT AAAAG T C AG T G C T G T AAAG T T GAAAGT T CAT GAGAGACATACAAT GAGGGC T GCAGCCCAT T T T TAAAAACAT TATAATACAAAA GTATGCACATTTGTTTACATATCCCTGCCTTTGTATTACAGTGGCAGGTTTGTGTACTTAAACT GGGAAAGCCTCAGATCTATGATTACCTGGCCTATCATAGAAAGTGTCTAAATAAATCACTCTGT CAAT T GAAT AC AT TAGTATTAGCTAGCATACTTCATTATGCCTGTTTTC CAT AAAT AC CAC AC C AAAAACTTGCTTGGGGCAGTTTGAGCCTAGTTCATGAGCTGCTATCAGATTGGTCTTGATCCTA TATAATAGGCCAAATGTCTGTAAACAGCTGTGCTGGTGGAATGTAGAAAGTCACTGCACTCAGA T T CAAC TTCCTGATTG GAAG T C AT CAC AG T G T GAT T AAAC AT T T T C AC AAAGAAT AG T AGAT AA ATAACTTGGTTTTTAATGTTAACTTTGTTTCCATTAAGTCACATTTAAAAACTTATCCTCACGC CTACCTGAGTTAATTATCTGTTGACCTAGATATCTTTCTGGCCACTCACTGACTTATTTCTTGA ACTTTTGCCATTTGCATAAATCTTGTCAGCTTTGTTCTTGATTATGCATTGTCCAGGCTGAGCT AGTTGTCTTTCCAGGAATCCCTTTGTCTCTGAATTAGGTCCTTTGTTTCCTAAATCATCCTGCT TGTTTGGCACAAGTCTTCCCAGGCCAGTGAGACCTCCGTGTCCTCTCAGCACCATAGGGGTAGG TAACCCTGGTTAGGCTGGACAGGGGTTTGCTGAGGGAGTTTGTTCATTTGAATCTAGGTCTTAC ATGACGTCTTTCAAATAGGGTTTTTACCTTGACACTAAACTGTCCAGTCTAAGCAGTTCTGCAA AATGTGAGGGAATTATGAACTTCTTCCTGCAGTGGGTTTTTATGGTTTTGGTTTGTTTTTTGTT GTTTTGGTTCTTTGTTGAGCCCTGGACAAAAACTTCCCTAGTTCTGGTTTCTACAATTTAAATT AAAAAC AGAAT T CAT C T TAGAAT T T T T CAC C C T C T T C C C CAAC TAT T C T AAT CAAT C T TAAG TATGCCCTTCATCTTTTTTCCTTCCTAAGGCTTTTACTGATAGTGTAATTCCGTACTCTTCAACCC TGGGAAGGCTGAAGTGGATTCTTGAGCTCATTTCAAGGCTGACCTGGGTGTTGGCAAGAACCCA GCTTAGAACAAACACATGCAAGGCCATCTTACCTTACATCCTGTTGCTTGGACTTCTTCCTGCT CAAAGTTTTTAGTGGATGCTAAGTGATCTTTGCTTCCACTGAGGAGTGGAACACTTTAGAATGA ACCTCTAGATAGATATTTTTATTGTCTGGTGAGGGTTACTGGAGTTTCCCACCCTGCCTGAAGG GTGAATCTGGCTTACAGTGTTCTCATCTCAAAGGGAAGAAGGCAGATGGCTGTGTCCAGAGAGA GCCATCACAGTTTGCTTCAGAGACACTAGAATGGGCTGGAAGATCTAGTGGTCTTAATCAGACT TGAAACCTGGCCTTTCTTCATTACCCATATGTCTACCAGTACTTGGGCTAACACTTAAGCCATT AGGGCCTTTGTAGGGGTGTTTTGAGACCCCCTCCATGCTAACAAATATACAGGTTTCTTAACAT TTGCTCATAAACTTGTAAAGCTTACTTTCTCTTAATCCACCCCACATTTAACAAGCCCTGGTAC TTAGAATTTCAGAAGAGTAATGGCAGGTAGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGT GTGTGTGT GAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGA AG T T T G T G GAAAAT GAG G T AAT GAG AG CTCATCCTTT T AGAAT T G T AC T T C AGAAT AGAAAC AT TTGGTGGGCTGTTAGGTAGCTTTGATTACTTGTGGGTAGACCTGCTAGTATTGCCAGTCCTCAA GCAATGAGCTTTCTGTATCTTGTTTACTAGATATATACTACCAGGTGAGTCATTTCCTGGGGTT CTGTTTTCTTTTAAAATCTTTCCCTAAACTTAATATGTATTAAAAAGTCTGGCTTTTCAGTCCA TTCTTTGTGCACTGGGATGGCAATTGCTTCATTATATGACAATTGCTGTTCCCAAGTCAGAATT CAGTGTGCTGATTTGACATCAGTTCGTCCCGAATAAGTTCCTGTTACCAGGATTTACATTCAGC ACATTAGAAACTTGTTGGTGTGCTTTTATTCTTGGAGCATTTTCCTTAGACTACCTTCCACTTT GAGTGCTCTGTTTAGGATGTTGAGGTGTTAGGATTCTTGACAGCCAGAAAGACTGAACCCACTA TCTGGGCACAGTGTTCGTGTTGCTCTATAAATGTATGCTTTTTTTGATTTGGGGTTGTTTTACC TACATTGTCAAACTAGATCCATGCTTAACAGTGATAATGAAGGCTTTTTGTTTGTTTTGTTTGT GGGTCCTCCCCCCCCCCCCAAGACAGGGTTTCTCTGTAGGCTGTCCTAGAACTTGTTCTTTTTT AACCAAAAT T T GGCAAGGC T GAAAAT GGAAT CC TATAAT CAAT GC T GGCCACAT TAAAGT TAAT AGTTGAGAAGTCTTGTCTGAATTTCCTTGGGCAAAAAGATTCTAGCCAGTTCAATACCCTGTTG TGCAAATTCAATTTGCTGTTATAATTTGCTCTCAGTTATCAGTTGGAAGGAGGTTAATTCTAATGTACTTGGAAGAGGCCTGTAGACCATCTATAACTGCATCAGTTGTACAGCGTTGTTGCCTGGGA TTCTCTAGTTCACATAAACTCCCAAGTCTTAGCCGTGGTGATGGCTACAGTGTGGAAGATGGTG AGCATTCTAGTGAGTATCGCGATGACGGCAGTAAAGAGCAGCAGGCAGCCGTGGCTGGGCTCAC TGACCGTGGCTGTAAGTTACGGAGGCAGCACACACTTCTGTACACACCTCTCATCAGTTACCGG AGTCATTGCATTGCGGACTAACTGGCTGACTCAAGTTGTCTTGCTACTGAAGTCTTGAGTTGGT CTCATGCATTTACCCTGTTGACTTGAGCACCTTAAAGTCGAAAGGATGTCTGGTTGTGGCTTTA TTGTAAACAGCCTTAGGTAAAGAGGGGAGTATATCGGTTAGGAAGGTGAAAAATGATACTTCCA AGTTCAGTGGGAAACCCTGGGTTTATCCCCCAGCTTAAGAAAGAATGCCTAACAATGTTTCAGA ATTAGATTCTGTGGAAGGTGAGGGTGTTAGAACAGTCCAAATTTGTTATTGTAGACTTGCAGTGGGAGGAAT T T T T AAAT AT ACAGAT GAG T C GACAC T CAT T AAC T T GAG T GAT AAAGG T GGAAAC G GATGTGGCAACACTTCTAAGTTCATTTGTATATGTTTGTAATTTGATTGGTTGTATTCTGTTGC ACTCTAGAATTTGAAGGCAAGGTTACCTCTGCTTTTTAATTTTTTTTTTTTTAAAGAAAGAAAA AAC AC T GAAAGAAAC T T C AAAAGAT CTGTTAATGCTAATACCT GAAT G T G G C AT T T AAC AT G T C AT G GAAAC T G C T T T GAAT AAAT AC T T GAGAAAAG GAAT GAAAT AAT T G C C G T T T T T G T T G T T GA GTGAATGGGTGTGGTTTAATGAGCGTAATCATTTTTATAAAACAGCTGTGAGACTGAAGTGGAA T C C T T AT T AAAT G T G GAAAAT G G C C T T T GAG GAT TAG AG T AGAGAT T C AAC T AAGAGAG T AAAT AAAGC T T GAAAC TAAT T CGT T GTAAAT T GC T T C TACAAT CAT T GC T C TATATAGCAT GC TAT T G CCAATCAGTTTTATGTATTAAGACCTATCAGCATGTCTTTTTTAGGTTGACCTCATTTTAAATT ATAAGATGCTCTCTGTACCGTTTTAACATTTCCAGGATTTATTCTTTCTAGGCAAATTCCACTG GACTGTTTCCATTGTAGAAGCTTCCTTATAGATTCTTCAAATGAAGCTTACAGTGTGCTTTCTT GGGGTTTTGATTTGCACTAAATTTTATTTTCTGAAAGATCACTTATGTTTATAATGTAGTGCTT T G T C T T AACAAT T AAAC T T T C CAGCAC T CAT GCA

[0052] The mouse p42AUF1amino acid sequence of GenBank Accession No.NP_001070734.1 (SEQ ID NO:20) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAAAAQGPAAAAGSGSGGGGSAAGGTEGGSAEAE GAKIDASKNEEDEGKMFIGGLSWDTTKKDLKDYFSKFGEWDCTLKLDPI TGRSRGFGFVLFKE SESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPDTPEEKIREYFGGFGEVES I ELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKVAMSKEQYQQQQQWGSRGGF AGRARGRGGGPSQNWNQGYSNYWNQGYGNYGYNSQGYGGYGGYDYTGYNNYYGYGDYSNQQSGY GKVSRRGGHQNSYKPY

[0053] The mouse p45AUF1nucleotide sequence of GenBank Accession No.NM_001077265.2 (SEQ ID NO:23) is as follows:CCATTTTAGGTGGTCCGCGGCGGCGCCATTAAAGCGAGGAGGAGGCGAGAGTGGCCGCCGCTGC TACTTCATTCTTTTTTTTTTCAGTGCAGCCGGGGAGAGCGAGAGAGCGCGCTGCGCGAGAGTGG GAGGCGAGGGGGGCAGGCCGGGGAGAGGCGCAGGAGCCCTTGCAGCCACGCGCGCGCCTTGTCT AGGGTGCCTCGCGAGGTAGAGCGGGCATCGCGCGGCGGCGGCGGGGATTACTTTGCTGCTAGTT TCGGTTCGCGGCGGCGGCGGCGTCGGCGGGTGTCGTCTTCGGCGGCGGCAGTAGCACTATGTCG GAGGAGCAGTTCGGAGGGGACGGGGCGGCGGCGGCGGCAACGGCGGCGGTAGGCGGCTCGGCGG GCGAGCAGGAGGGAGCCATGGTGGCGGCGGCGGCGCAGGGGCCGGCGGCGGCGGCGGGAAGCGG GAGCGGCGGCGGCGGCTCTGCGGCCGGAGGCACCGAAGGAGGCAGCGCCGAGGCAGAGGGAGCC AAGATCGACGCCAGTAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACACACTG AAGCAGCGGCGGCACAGCGGGAAGAATGGAAAATGTTTATAGGAGGCCTTAGCTGGGACACCAC AAAGAAAGAT C T GAAG GAC T AC T T T T C C AAAT T T G G T GAAG T T G T AGAC T G C AC T C T GAAG T T AGATCCTATCACAGGGCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAGTCGGAGAGTGTAG ATAAGGT CAT GGAT CAGAAAGAACATAAAT T GAAT GGGAAAGT CAT T GAT CC TAAAAGGGCCAA AGCCATGAAAACAAAAGAGCCTGTCAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGACACACCT GAAGAAAAAATAAGAGAGTACTTTGGTGGTTTTGGTGAGGTTGAATCCATAGAGCTCCCTATGG ACAACAAGACCAATAAGAGGCGTGGGTTCTGTTTTATTACCTTTAAGGAAGAGGAGCCAGTGAA GAAGAT AAT G GAAAAGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG C C AT G TCAAAGGAACAGTATCAGCAGCAGCAGCAGTGGGGATCTAGAGGAGGGTTTGCAGGCAGAGCTC GCGGAAGAGGTGGAGGCCCCAGTCAAAACTGGAACCAGGGATATAGTAACTATTGGAATCAAGG CTATGGCAACTATGGATATAACAGCCAAGGTTACGGAGGTTATGGAGGATATGACTACACTGGT TACAACAACTACTATGGATATGGTGATTATAGCAATCAGCAGAGTGGTTATGGGAAAGTATCCA GGCGAGGTGGACATCAAAATAGCTACAAACCATACTAAATTATTCCATTTGCAACTTATCCCCA ACAGGTGGTGAAGCAGTATTTTCCAATTTGAAGATTCATTTGAAGGTGGCTCCTGCCACCTGCT AATAGCAGTTCAAACTAAATTTTTTCTATCAAGTTCCTGAATGGAAGTATGACGTTGGGTCCCT CTGAAGTTTAATTCTGAGTTCTCATTAAAAGAATTTGCTTTCATTGTTTTATTTCTTAATTGCT ATGCTTCAGTATCAATTTGTGTTTTATGCCCCCCCTCCCCCCCAGTATTGTAGAGCAAGTCTTG TGTTAAAAAAAGCCCAGTGTGACAGTGTCATGATGTAGTAGTGTCTTACTGGTTTTTTAATAAA TCCTTTTG T AT AAAAAT G TAT TGGCTCTTT TAT CAT CAGAAT AGGAGGAAG T GAAAT AC T ACAA ATGTTTGTCTTGGATTCAAGTCACTAGAAGCATAAATTTGAGGGGATAAAAACAACGGTAAACT TTGTCTGAAAGAGGGCATGGTTAAAAATGTAGTGAATTTTAAATGTTTTTAGCAAAATTTGATT TTGCCCAAGAATCCCTGTCTGAATTGGAAATGACTTAATGTAGTCAATGTGCTTGTTGGTTGTC TTAATATTACTTCTGTAGCCATTAAGTTTTATGAGTAACTTCCCAAATACCCACGTTTTTCTTT ATAT GTAT TGTGCTTTT TAAAAACAAAT C T GGAAAAAT GGGCAAGAACAT T T GCAGACAAT T GT T T T T AAGC T T C CAT T AAAT AAAAAAAAT G T GGAC T T AAGGAAAT C TAT T AAT T T AAAT AGAAC T GCAGC TAGT T TAGAGAGTAT TTTTTTCT TAAAGC T T T GGT GTAAT TAGGGAAGAT T T TAAAAAA TGCATAGTGTTTATTTGTATGTGTGCTCTTTTTTTAAGTCAATTTTTGGGGGGTTGGTCTGTTA ACTGAGTCTAGGATTTAAAGGTAAGATGTTCCTAGAAATCTTGTCATCCCAAAGGGGCGGGCGC TAAGGTGAAACTTCAGGGTTCAGTCAGGGTCACTGCTTTATGTGTGAAATCACTCAAATTGGTA AGT C T C T TAT GT TAGCAT T CAGGACAT T GAT T T CAAC T T GGAT GGACAAT T TATAGT TAG TAG T GAATTGTGTGTTAATGTGTTCAGTCCTGGTAAGTTTTCAGTTTGATCAGTTAGTTGGAAGCAGA CTTGAAGAGCTGTTAGTCACGTGAGCCATGGGTGCAGTCGATCTGTGGTCAGATGCCTGAGTCT G T GAT AG T GAAT T G T G T C TAAAGAC AT T T T AAT GAT AAAAG T GAG T G C T G TAAAG T T GAAAG T T CAT GAGAGACATACAAT GAGGGC T GCAGC GGAT T T T TAAAAACAT TATAATACAAAAGTAT GCA CATTTGTTTACATATCCCTGCCTTTGTATTACAGTGGCAGGTTTGTGTACTTAAACTGGGAAAG C C T C AGAT GTAT GATT AC CTGGCCTAT C AT AGAAAG T G T C T AAAT AAAT GAG T C T G T GAAT T GA AT AC AT TAGTATTAGCTAGCATACTTCATTATGCCTGTTTTC CAT AAAT AC GAG AC C AAAAAC TTGCTTGGGGCAGTTTGAGCCTAGTTCATGAGCTGCTATCAGATTGGTCTTGATCCTATATAATA GGCCAAATGTCTGTAAACAGCTGTGCTGGTGGAATGTAGAAAGTCACTGCACTCAGATTCAACT TCCTGATTG GAAG T C AT GAG AG T G T GAT T AAAC AT T T T C AC AAAGAAT AG T AGAT AAAT AAC T T GGTTTTTAATGTTAACTTTGTTTCCATTAAGTCACATTTAAAAACTTATCCTCACGCCTACCTG AGTTAATTATCTGTTGACCTAGATATCTTTCTGGCCACTCACTGACTTATTTCTTGAACTTTTG CCATTTGCATAAATCTTGTCAGCTTTGTTCTTGATTATGCATTGTCCAGGCTGAGCTAGTTGTC TTTCCAGGAATCCCTTTGTCTCTGAATTAGGTCCTTTGTTTCCTAAATCATCCTGCTTGTTTGG CACAAGTCTTCCCAGGCCAGTGAGACCTCCGTGTCCTCTCAGCACCATAGGGGTAGGTAACCCT GGTTAGGCTGGACAGGGGTTTGCTGAGGGAGTTTGTTCATTTGAATCTAGGTCTTACATGACGT CTTTCAAATAGGGTTTTTACCTTGACACTAAACTGTCCAGTCTAAGCAGTTCTGCAAAATGTGA GGGAATTATGAACTTCTTCCTGCAGTGGGTTTTTATGGTTTTGGTTTGTTTTTTGTTGTTTTGG TTCTTTGTTGAGCCCTGGACAAAAACTTCCCTAGTTCTGGTTTCTACAATTTAAATTAAAAACA GAATTCATCTTAGAATTTTTCACCCTCTTCCCCAACTATTCTAATCAATCTTAAGTATGCCCTT CATCTTTTTTCCTTCCTAAGGCTTTTACTGATAGTGTAATTCCGTACTCTTCAACCCTGGGAAG GCTGAAGTGGATTCTTGAGCTCATTTCAAGGCTGACCTGGGTGTTGGCAAGAACCCAGCTTAGA ACAAACACATGCAAGGCCATCTTACCTTACATCCTGTTGCTTGGACTTCTTCCTGCTCAAAGTT TTTAGTGGATGCTAAGTGATCTTTGCTTCCACTGAGGAGTGGAACACTTTAGAATGAACCTCTA GATAGATATTTTTATTGTCTGGTGAGGGTTACTGGAGTTTCCCACCCTGCCTGAAGGGTGAATC TGGCTTACAGTGTTCTCATCTCAAAGGGAAGAAGGCAGATGGCTGTGTCCAGAGAGAGCCATCA CAGT T T GC T T CAGAGACAC TAGAAT GGGC T GGAAGAT C TAGT GGT C T TAAT CAGAC T T GAAACC TGGCCTTTCTTCATTACCCATATGTCTACCAGTACTTGGGCTAACACTTAAGCCATTAGGGCCT TTGTAGGGGTGTTTTGAGACCCCCTCCATGCTAACAAATATACAGGTTTCTTAACATTTGCTCA T AAAC T T G TAAAG C T TAG T T T C T C T TAAT C GAG C C GAG AT T T AAC AAG C C C T G G TAG T TAGAAT TTCAGAAGAGTAATGGCAGGTAGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG T GAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAAG T T T G T GGAAAAT CAGGTAAT GACAGC T CAT CC T T T TAGAAT T GTAC T T CAGAATAGAAACAT T T GGT GG GCTGTTAGGTAGCTTTGATTACTTGTGGGTAGACCTGCTAGTATTGCCAGTCCTCAAGCAATGA GCTTTCTGTATCTTGTTTACTAGATATATACTACCAGGTGAGTCATTTCCTGGGGTTCTGTTTT CTTTTAAAATCTTTCCCTAAACTTAATATGTATTAAAAAGTCTGGCTTTTCAGTCCATTCTTTG TGCACTGGGATGGCAATTGCTTCATTATATGACAATTGCTGTTCCCAAGTCAGAATTCAGTGTG C T GAT T T GAC AT C AG TTCGTCCC GAAT AAG TTCCTGTTAC C AG GATT TAG AT T C AG C AC AT TAG AAACTTGTTGGTGTGCTTTTATTCTTGGAGCATTTTCCTTAGACTACCTTCCACTTTGAGTGCT CTGTTTAGGATGTTGAGGTGTTAGGATTCTTGACAGCCAGAAAGACTGAACCCACTATCTGGGC ACAGTGTTCGTGTTGCTCTATAAATGTATGCTTTTTTTGATTTGGGGTTGTTTTACCTACATTG TCAAACTAGATCCATGCTTAACAGTGATAATGAAGGCTTTTTGTTTGTTTTGTTTGTGGGTCCTCCCCCCCCCCCCAAGACAGGGTTTCTCTGTAGGCTGTCCTAGAACTTGTTCTTTTTTAACCAAA ATTTGGCAAGGCTGAAAATGGAATCCTATAATCAATGCTGGCCACATTAAAGTTAATAGTTGAG AAGTCTTGTCTGAATTTCCTTGGGCAAAAAGATTCTAGCCAGTTCAATACCCTGTTGTGCAAAT TCAATTTGCTGTTATAATTTGCTCTCAGTTATCAGTTGGAAGGAGGTTAATTCTAATGTACTTG GAAGAGGCCTGTAGACCATCTATAACTGCATCAGTTGTACAGCGTTGTTGCCTGGGATTCTCTA GTTCACATAAACTCCCAAGTCTTAGCCGTGGTGATGGCTACAGTGTGGAAGATGGTGAGCATTC TAGTGAGTATCGCGATGACGGCAGTAAAGAGCAGCAGGCAGCCGTGGCTGGGCTCACTGACCGT GGCTGTAAGTTACGGAGGCAGCACACACTTCTGTACACACCTCTCATCAGTTACCGGAGTCATT GCATTGCGGACTAACTGGCTGACTCAAGTTGTCTTGCTACTGAAGTCTTGAGTTGGTCTCATGC ATTTACCCTGTTGACTTGAGCACCTTAAAGTCGAAAGGATGTCTGGTTGTGGCTTTATTGTAAA CAGCCTTAGGTAAAGAGGGGAGTATATCGGTTAGGAAGGTGAAAAATGATACTTCCAAGTTCAG TGGGAAACCCTGGGTTTATCCCCCAGCTTAAGAAAGAATGCCTAACAATGTTTCAGAATTAGAT TCTGTGGAAGGTGAGGGTGTTAGAACAGTCCAAATTTGTTATTGTAGACTTGCAGTGGGAGGAA T T T T T AAAT AT AC AGAT GAG T C GAG AC T C AT T AAC T T C AC T GAT AAAG G T G GAAAC G GAT G T G G CAACACTTCTAAGTTCATTTGTATATGTTTGTAATTTGATTGGTTGTATTCTGTTGCACTCTAG AAT T T GAAGGCAAGGT TACC T C T GC T T T T TAAT T T T T T T T T T T T TAAAGAAAGAAAAAACAC T G AAAGAAAC T T CAAAAGAT C T G T T AAT GC TAAT AC C T GAAT G T GGCAT T T AACAT G T CAT GGAAA CTGCTTTGAATAAATACTTGAGAAAAGGAATGAAATAATTGCCGTTTTTGTTGTTGAGTGAATG GGTGTGGTTTAATGAGCGTAATCATTTTTATAAAACAGCTGTGAGACTGAAGTGGAATCCTTAT TAAAT GT GGAAAAT GGCC T T T GAGGAT TACAGTAGAGAT T CAAC TAAGAGAG T AAAT AAAG C T T GAAAC TAAT T CGT T GTAAAT T GC T T C TACAAT CAT T GC T C TATATAGCAT GC TAT T GCCAAT CA GTTTTATGTAT T AAGAC C T AT C AG CATGTCTTTTTTAGGTT GAC C T C AT T T TAAAT T AT AAGAT GCTCTCTGTACCGTTTTAACATTTCCAGGATTTATTCTTTCTAGGCAAATTCCACTGGACTGTT TCCATTGTAGAAGCTTCCTTATAGATTCTTCAAATGAAGCTTACAGTGTGCTTTCTTGGGGTTT TGATTTGCACTAAATTTTATTTTCTGAAAGATCACTTATGTTTATAATGTAGTGCTTTGTCTTA ACAAT TAAAC T T T CCAGCAC T CAT GCA

[0054] The mouse p45AUF1amino acid sequence of GenBank Accession No.NP_001070733.1 (SEQ ID NO:24) is as follows:MSEEQFGGDGAAAAATAAVGGSAGEQEGAMVAAAAQGPAAAAGSGSGGGGSAAGGTEGGSAEAE GAKIDASKNEEDEGHSNSSPRHTEAAAAQREEWKMFIGGLSWDTTKKDLKDYFSKFGEWDCTL KLDPI TGRSRGFGFVLFKESESVDKVMDQKEHKLNGKVIDPKRAKAMKTKEPVKKI FVGGLSPD TPEEKIREYFGGFGEVES IELPMDNKTNKRRGFCFI TFKEEEPVKKIMEKKYHNVGLSKCE IKV AMSKEQYQQQQQWGSRGGFAGRARGRGGGPSQNWNQGYSNYWNQGYGNYGYNSQGYGGYGGYDY TGYNNYYGYGDYSNQQSGYGKVSRRGGHQNSYKPY

[0055] It is noted that the sequences described herein may be described with reference to accession numbers that include, e.g., a coding sequence or protein sequence with or without additional sequence elements or portions (e.g., leader sequences, tags, immature portions, regulatory regions, etc.). Thus, reference to such sequence accession numbers or corresponding sequence identification numbers refers to either the sequence fully described therein or some portion thereof (e.g., that portion encoding a protein or polypeptide of interest to the technology described herein (e.g., AUF1 or a functional fragment thereof); the mature protein sequence that is described within a longer amino acid sequence; a regulatory region of interest (e.g., promoter sequence or regulatory element) disclosed within a longer sequence described herein; etc). Likewise, variants and isoforms of accession numbers and corresponding sequence identification numbers described herein are also contemplated.

[0056] Accordingly, in certain embodiments, the AUF1 protein referred to herein has an amino acid sequence as set forth in Table 1 and the sequences disclosed herein, or is a functional fragment thereof. In one embodiment, the functional fragment as referred to herein includes an amino acid sequence that has at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or at least 99% amino acid sequence identity to an amino acid sequence disclosed herein.

[0057] The subject may have: (i) degeneration of neuromuscular junctions; (ii) a genetic neuromuscular wasting disorder, optionally wherein the genetic neuromuscular wasting disorder is amyotrophic lateral sclerosis (ALS), Lambert-Eaton Myasthenic Syndrome (LEMS), or a muscular dystrophy, optionally wherein the muscular dystrophy is spinal muscular atrophy (SMA); (iii) a myopathy with a neurologic or inflammatory disorder, optionally wherein the myopathy with a neurologic or inflammatory disorder is inclusion body myositis, Myasthenia gravis, neurogenic amyloidosis, Sjogren's syndrome, multiple sclerosis (MS), ataxia, or rheumatoid arthritis; (iv) an autoimmune disorder, optionally wherein the autoimmune disorder is myasthenia gravis (MG) or Lambert Eaton syndrome; (v) a nerve trauma injury, a nerve cut, or a compression-decompression injury; (vi) a loss of neuromuscular junction as a result of a neurogenic myopathy, optionally wherein the neurologic myopathy is a vacuolar myopathy such as bulbospinal muscular atrophy; and / or (vii) a loss of neuromuscular junctions as a result of a muscular dystrophy, optionally wherein the muscular dystrophy is spinal muscular atrophy (SMA).

[0058] In some embodiments, the subject has degeneration of neuromuscular junctions.

[0059] In some embodiments, the subject has a neurogenic myopathy.

[0060] In some embodiments, the subject has age induced muscle mass loss or sarcopenia.

[0061] Age related and sarcopenic muscle loss are well established to involve loss of NMJs, and given muscle weakness, may be assumed even in the absence of biopsy. In some embodiments, age induced muscle mass loss or sarcopenia may be measured by muscle biopsy (Moreiera-Pais et al, “A Neuromuscular Perspective of Sarcopenia Pathogenesis: Deciphering the Signaling Pathways Involved,” GeroScience 44: 1199-1213 (2022); Arnold and Clark, “Neuromuscular Junction Transmission Failure in Aging and Sarcopenia: The Nexus of the Neurological and Muscular Systems,” Aging Research Reviews 89: 101966 (2023), which are hereby incorporated by reference in their entirety), staining the biopsy specimen for acetylcholine receptors (AChRs) using specific antibodies, and for the state of denervation using specific antibodies to neural cell adhesion molecule (NCAM) see, e.g., Soendenbroe et al., “Muscle-Nerve Communication and the Molecular Assessment of Human Skeletal Muscle Denervation with Aging,” Am. J. Physiol. Cell Physiol. 32EC317-C329 (2021), which is hereby incorporated by reference in its entirety). Various muscle fiber electrophysiology tests including repetitive nerve stimulation, clinical needle electromyo-graphy, and electromyographic motor unit measurements can also be used to assess NMJ transmission ability (Arnold and Clark, “Neuromuscular Junction Transmission Failure in Aging and Sarcopenia: The Nexus of the Neurological and Muscular Sy stems,” Aging Research Reviews 89: 101966 (2023), which is hereby incorporated by reference in its entirety).

[0062] In some embodiments, the subject has a genetic neuromuscular wasting disorder. Genetic neuromuscular wasting disorders are conditions caused by genetic mutations that lead to the progressive degeneration and weakening of muscles and nerves that control them. The disorders are typically characterized by muscle atrophy, weakness, and loss of motor function.

[0063] In some embodiments, the genetic neuromuscular wasting disorder is amyotrophic lateral sclerosis (ALS), Lambert-Eaton Myasthenic Syndrome (LEMS), or a muscular dystrophy.

[0064] Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder primarily affecting the motor system, characterized by the progressive loss of upper and lower motor neurons in the motor cortex, brain stem nuclei, and anterior horn of the spinal cord, leading to muscle weakness and wasting, with increasingly recognized extra-motor manifestations (Masrori and Van Damme, “Amyotrophic Lateral Sclerosis: A Clinical Review,” Eur. J. Neurol.27(10): 1918-1929 (2020), which is hereby incorporated by reference in its entirety). In some embodiments, the genetic neuromuscular wasting disorder is amyotrophic lateral sclerosis (ALS).

[0065] Lambert-Eaton Myasthenic Syndrome (LEMS) is a paraneoplastic or primary autoimmune neuromuscular junction disorder characterized by proximal weakness and autonomic dysfunction (Kesner et al., “Lambert-Eaton Myasthenic Syndrome,” Neurol. Clin. 36(2):379-394 (2019), which is hereby incorporated by reference in its entirety). The characteristic weakness is thought to be caused by antibodies targeting P / Q-type voltage-gated calcium channels (VGCC) on presynaptic nerve terminals and resulting in diminished release of acetylcholine (ACh) (Kesner et al., “Lambert-Eaton Myasthenic Syndrome,” Neurol. Clin. 36(2):379-394 (2019), which is hereby incorporated by reference in its entirety). In some embodiments, the genetic neuromuscular wasting disorder is Lambert-Eaton Myasthenic Syndrome (LEMS).

[0066] Muscular dystrophies encompass a group of genetic conditions with progressive muscle damage and weakness (Hoang and Dowdy, “A Review of Muscular Dystrophies,” Anesth. Prog. 71 (1) :44-52 (2024), which is hereby incorporated by reference in its entirety). As used herein, the term “muscular dystrophy” includes, for example, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), Limb-girdle muscular dystrophy, Congenital muscular dystrophy, Facioscapulohumeral muscular dystrophy, Myotonic muscular dystrophy, Oculopharyngeal muscular dystrophy, Distal muscular dystrophy, and Emery- Dreifuss muscular dystrophy. In some embodiments, the muscular dystrophy is characterized, at least in part, by a deficiency or dysfunction of the protein dystrophin. Such muscular dystrophies may include Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (DMD). In some embodiments, the muscular dystrophy is associated with degenerative muscle conditions such as muscle disuse atrophy, denervation muscle atrophy, dysferlinopathy, AIDS / HIV, diabetes, chronic obstructive pulmonary disease, kidney disease, cancer, aging, autoimmune disease, polymyositis, and dermatomyositis.

[0067] In some embodiments, the muscular dystrophy is spinal muscular atrophy (SMA), facioscapulohumeral muscular dystrophy (FSHD), Emery-Dreifuss muscular dystrophy (EDMD), Duchenne muscular dystrophy (DMD), limb girdle muscular dystrophy (LGMD), myotonic dystrophy, or congenital muscular dystrophy.

[0068] Spinal muscular atrophy (SMA) is a neurodegenerative disorder caused by mutations in SMN1, which encodes survival motor neuron protein (SMN), and reduced expression of SMN leads to the loss of a-motor neurons and severe muscle weakness (Mercuri et al., “Spinal Muscular Atrophy,” Nature 8:52 (2022), which is hereby incorporated by reference in its entirety). In some embodiments, the muscular dystrophy is spinal muscular atrophy (SMA).

[0069] In some embodiments, the subject has a myopathy with a neurologic or inflammatory disorder. Myopathies are a heterogenous group of disorders primarily affecting the skeletal muscle structure, metabolism, or channel function. Myopathies may be inherited or acquired.

[0070] Exemplary inherited myopathies include, without limitation, mitochondrial myopathies, congenital myopathies, metabolic myopathies, channelopathies, and muscular dystrophies.

[0071] Exemplary acquired myopathies include, without limitation, toxic myopathies, immune-mediated or idiopathic inflammatory myopathies, infectious myopathies (such as myopathies associated with bacterial infections, viral infections, parasitic infections, and fungal infections), endocrine myopathies (such as myopathies associated with thyroid and parathyroid dysfunction, adrenal dysfunction, and diabetic muscle infarction), electrolyte-mediated myopathies, and myopathies associated with systemic disease. Toxic myopathies include antimicrotubular myopathies such as myopathy associated with vincristine. Vincristine is a vital constituent of chemotherapeutic regimens. Vincristine-induced neuropathy is a challenging adverse effect that impacts quality of life and treatment course (Alwhaibi et al., “Vincristine- Induced Neuropathy in Patients Diagnosed with Solid and Hematological Malignancies: The Role of Dose Rounding,” J. Clin. Med. 12(17):5662 (2023), which is hereby incorporated by reference in its entirety). Additional toxic myopathies include, without limitation, amphiphilic myopathies associated with chloroquine, hydroxychloroquine, and amiodarone (Pasnoor et al., “Toxic Myopathies,” Neurol. Clin. 32(3) :647-viii (2014), which is hereby incorporated by reference in its entirety), necrotizing myopathies (associated with statins, fibrates, immune checkpoint inhibitors, labetalol, propofol, alcohol, and cyclosporine), mitochondrial myopathies (associated with some antiretrovirals), hypokalemic myopathies (associated with diuretics, steroids, laxatives, alcohol, etc.), critical care-associated myopathies (associated with corticosteroids and neuromuscular blockers), inflammatory myopathy (associated with tumor necrosis factor alfa inhibitors, immune checkpoint inhibitors, statins, interferon-alfa, D- penicillamine, L-tryptophan, hydroxyurea, imatinib lamotrigine, and phenytoin).

[0072] In some embodiments, the myopathy with a neurologic or inflammatory disorder is Polymyositis, dermatomyositis, inclusion body myositis, Myasthenia gravis, neurogenic amyloidosis, or Sjogren's syndrome.

[0073] Inclusion body myositis (IBM) is an idiopathic inflammatory myopathy which affects proximal and distal muscles, causing weakness of the quadriceps and finger flexor muscles (Chardon et al., “Inclusion Body Myositis,” CMAJ 196(14):E486 (2024), which ishereby incorporated by reference in its entirety). In some embodiments, the myopathy with a neurologic or inflammatory disorder is inclusion body myositis.

[0074] Myasthenia gravis is an antibody-mediated autoimmune disease which is dependent on T cells and characterized by the presence of autoantibodies targeting proteins located on the postsynaptic surface of skeletal muscle, known as the neuromuscular junction (Kaminski et al., “Myasthenia Gravis: The Future is Here,” J. Clin. Invest. 134(12):el79742 (2024), which is hereby incorporated by reference in its entirety). Autoimmune destruction of the neuromuscular junctions (NMJs) that transmit motor neuron impulses to muscle fibers causes weakness in voluntary muscles that varies widely in severity and scope among affected individuals (Kaminski et al., “Myasthenia Gravis: The Future is Here,” J. Clin. Invest.134(12):el79742 (2024), which is hereby incorporated by reference in its entirety). In some embodiments, the myopathy with a neurologic or inflammatory disorder is myasthenia gravis.

[0075] Amyloidosis is a condition involving deposits of protein as a result of inflammatory conditions. In some embodiments, the myopathy with a neurologic or inflammatory disorder is neurogenic amyloidosis.

[0076] Sjogren's syndrome is a chronic autoimmune connective tissue disease characterized by sicca symptoms, fatigue, and pain (Andre and Bbckle, “Sjogren's Syndrome,” J. Dtsch. Dermatol. Ges. 20(7):980-1002 (2022), which is hereby incorporated by reference in its entirety). In some embodiments, the myopathy with a neurologic or inflammatory disorder is Sjogren's syndrome.

[0077] In some embodiments, the subject has multiple sclerosis (MS), ataxia, or rheumatoid arthritis.

[0078] Multiple sclerosis (MS) is an autoimmune-mediated neurodegenerative disease of the central nervous system characterized by inflammatory demyelination with axonal transection (McGinley et al., “Diagnosis and Treatment of Multiple Sclerosis,” JAMA 325(8):765-779 (2021), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has multiple sclerosis.

[0079] Ataxia is a neurological sign that manifests in a lack of coordination in the movement of different muscles in the body. In some embodiments, the subject has ataxia.

[0080] Rheumatoid arthritis (RA) is an inflammatory systemic disease primarily affecting the joints and musculoskeletal structures, typically manifesting as polyarthritis due to autoimmune synovitis, with potential extra-articular manifestations such as interstitial lung disease or vasculitis affecting small vessels (Bauhammer and Fiehn, “Rheumatoid Arthritis,”DtschMed. Wochenschr. 150(9): 508-520 (2025), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has rheumatoid arthritis.

[0081] In some embodiments, the subject has an autoimmune disorder. The autoimmune disorder may be myasthenia gravis (MG) or Lambert Eaton syndrome. In some embodiments, when the subject has an autoimmune disorder, the subject produces autoantibodies against neuromuscular junctions (NMJs).

[0082] In some embodiments, when the subject has an autoimmune disorder, the subject produces autoantibodies. Autoantibodies include antibodies against the acetylcholine receptor (AChR), muscle specific kinase (MuSK), low density lipoprotein receptor-related protein (LRP4), and / or voltage gated calcium channels (VGCCs).

[0083] As noted above, acetylcholine receptors (AChRs) are heteromeric membrane proteins that cluster to ensure proper signal transduction and are essential for neurotransmission at the neuromuscular junction. The pathogenic mechanisms of acetylcholine receptor (AChR) autoantibodies, e.g., in myasthenia gravis (MG), include complement activation leading to destruction of the muscle membrane of the neuromuscular junction via reduction in postsynaptic membrane folds; antigen modulation causing accelerated internalization and degradation of AChRs; and functional blockade of ACh-AChR binding (Ma et al., “Advancements and Prospects of Novel Biologicals for Myasthenia Gravis: Toward Personalized Treatment Based on Autoantibody Specificities,” Front. Pharmacol. 15: 1370411 (2024), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has an autoimmune disorder and produces autoantibodies against acetylcholine receptors (AChRs).

[0084] Muscle-specific kinase (MuSK) is a single-subunit transmembrane protein located on the postsynaptic membrane of the neuromuscular junction that plays an important role in clustering the AChR (Gilhus et al., Myasthenia Gravis,” Nat. Rev. Dis. Prim. 5(1) :30 (2019), which is hereby incorporated by reference in its entirety). Under normal circumstances, agrin, which is synthesized by motor neurons, binds to the MuSK co-receptor LRP4 to trigger the activation of MuSK, thereby clustering the AChR and achieving the maintenance of the postsynaptic membrane (Ma et al., “Advancements and Prospects of Novel Biologicals for Myasthenia Gravis: Toward Personalized Treatment Based on Autoantibody Specificities,” Front. Pharmacol. 15: 1370411 (2024), which is hereby incorporated by reference in its entirety). The pathogenic mechanisms of MuSK autoantibodies, e.g., in myasthenia gravis (MG), include blocking the LRP4-MuSK interaction, thereby disrupting the agrin / MuSK signaling pathway and its maintenance of the integrity of the neuromuscular junction structure and function, resulting in impaired neuromuscular transmission and forming symptoms of muscle weakness;and inhibition of ACh vesicle clustering in motor nerve terminals by controlling presynaptic membrane development through the retrograde signal (Ma et al., “Advancements and Prospects of Novel Biologicals for Myasthenia Gravis: Toward Personalized Treatment Based on Autoantibody Specificities,” Front. Pharmacol. 15: 1370411 (2024), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has an autoimmune disorder and produces autoantibodies against muscle-specific kinase (MuSK).

[0085] LRP4 is a member of the low-density lipoprotein receptor (LDLR) family which comprises an extracellular N-terminal region that possesses multiple EGF repeats and LDLR repeats, a transmembrane domain, and a short C-terminal region (Shen et al., “Antibodies Against Low-Density Lipoprotein Receptor-Related Protein 4 Induce Myasthenia Gravis,” J. Clin. Invest. 123(12): 5190-5202 (2013), which is hereby incorporated by reference in its entirety). As described supra, LRP4 is a receptor of agrin critical for MuSK activation, AChR clustering, and neuromuscular junction formation. The pathogenic mechanisms of LRP4 autoantibodies, e.g., in myasthenia gravis (MG), include damage to the neuromuscular junction by interfering with agrin / MuSK signaling and complement fixation (Shen et al., “Antibodies Against Low-Density Lipoprotein Receptor-Related Protein 4 Induce Myasthenia Gravis,” J. Clin. Invest. 123(12): 5190-5202 (2013), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has an autoimmune disorder and produces autoantibodies against low density lipoprotein receptor (LRP4).

[0086] Voltage gated calcium channels (VGCCs) are multi-subunit proteins crucial for cell membrane depolarization and calcium influx in response to action potentials, functioning as secondary messengers in electrical signalization and initiating various cellular mechanisms (Bekircan-Kurt, “Voltage gated calcium channel antibody-related neurological diseases,” World J. Clin. Cases 3(3):293-300 (2015), which is hereby incorporated by reference in its entirety). As described herein, VGCCs are immunologic targets for several diseases, including Lambert- Eaton myasthenic syndrome (LEMS) (Hajela et al., “Lambert-Eaton Syndrome Antibodies Target Multiple Subunits of Voltage-Gated Ca2+ Channels,” Muscle andNerve 51(2): 17-184 (2015), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has an autoimmune disorder and produces autoantibodies against voltage gated calcium channels (VGCCs).

[0087] As noted above, myasthenia gravis is an antibody-mediated autoimmune disease, characterized by the presence of autoantibodies targeting proteins located on the postsynaptic surface of skeletal muscle, known as the neuromuscular junction (Kaminski et al., “Myasthenia Gravis: The Future is Here,” J. Clin. Invest. 134(12):el79742 (2024), which is herebyincorporated by reference in its entirety). Myasthenia gravis may be characterized, e.g., by the presence and / or absence of autoantibodies against acetylcholine receptors (AChRs), musclespecific tyrosine kinase (MusK), and lipoprotein receptor-related protein 4 (LRP4). Thus, in some embodiments, when the subject has myasthenia gravis, the subject may have AChR-MG, MuSK-MG, LRP4-MG or seronegative MG.

[0088] In some embodiments, the subject produces autoantibodies against muscle acetylcholine receptor (AChR), muscle specific kinase (MuSK), low density lipoprotein receptor-related protein 4 (LRP4), and / or voltage gated calcium channels (VGCCs).In some embodiments, the subject demonstrates muscle weakness due to aging or sarcopenia.

[0089] As used herein, “myopathy” is a generalized term referring to any disease concerning the muscles. Hallmarks of drug-induced myopathy include fatigue, generalized muscle pain, muscle tenderness, muscle weakness, (nocturnal) cramping, and tendon pain. A “drug-induced myopathy” or “toxic myopathy” is a subacute, and rarely acute, manifestation of myopathic symptoms, such as muscle weakness, fatigue, myalgia, cramps, creatine kinase (CK) elevation, or myoglobinuria, that occurs in patients without muscle disease when exposed to therapeutic doses of certain drugs (see, e.g., Janssen et al., “Muscle Toxicity of Drugs: When Drugs Turn Physiology into Pathophysiology,” Physiological Reviews 100(2):633-72 (2020), which is hereby incorporated by reference in its entirety).

[0090] In some embodiments, a subject has sustained loss of neuromuscular function as a result of chemical or drug toxicity (such as botulism toxin, cancer drug exposure such as vincristine). The chemical or drug toxicity may be a results of: (i) a pre-synaptic toxin such as botulism toxin or tetanus toxin or a post-synaptic snake venom; (ii) a neurotoxin such as a- bungarotoxin, a-cobratoxin, notexin, or taipoxin; (iii) certain antibiotics such as fluoroquinolone and natural quinoline derivatives such as quinine or quinidine; (iv) aminoglycoside antibiotics; and / or (v) cancer drugs such as vincristine.

[0091] Tetanus toxin blocks neurotransmitter release from spinal inhibitory interneurons after retroaxonal transport to the spinal cord, while botulinum toxin causes inhibits acetylcholine release at the neuromuscular junction (Silva et al., “Defining the Role of Post-Synaptic a- Neurotoxins in Paralysis Due to Snake Envenoming in Humans,” Cell Mol. Life Sci. 75(23): 4465-4478 (2018) and Pellizzari et al., “Tetanus and Botulinum Neurotoxins: Mechanism of Action and Therapeutic Uses,” Philos. Trans. R Soc. Lond. B. Biol. Sci. 354(1381):2589-268 (1999), which are hereby incorporated by reference in their entirety). In some embodiments, the sustained loss of neuromuscular function is a result of a pre-synaptic neurotoxin such as botulinum toxin and tetanus toxin.

[0092] Post-synaptic snake venoms are competitive antagonists that bind with high affinity to the two agonists binding sites of the adult muscle-type (al -5 and al -a interface of (al)2p5a) nicotinic acetylcholine receptors (nAChR) on the motor end plate, leading to neuromuscular blockade (Nirthanan and Gwee, “Three-Finger Alpha-Neurotoxins and the Nicotinic Acetylcholine Receptor, Forty Years On,” J. Pharmacol. Sci. 94(1): 1-17 (2004), which is hereby incorporated by reference in its entirety). In some embodiments, the sustained loss of neuromuscular function is a result of a post-synaptic snake venom such as a-bungarotoxin, a- cobratoxin, or a-neurotoxin (Osipov and Utkin, “What are the Neurotoxins in Hemotoxic Snake Venoms,” Int. J. Mol. Sci. 24(3):2919 (2023), which is hereby incorporated by reference in its entirety).

[0093] Additional suitable neurotoxins include, without limitation, pre-synaptic snake venom notexin and pre-synaptic snake venom taipoxin (Cull-Candy et al., “The Effects of Taipoxin and Notexin on the Function and Fine Structure of the Murine Neuromuscular Junction,” Neuroscience 1 (3):P 175- 180 (1976), which is hereby incorporated by reference in its entirety). In some embodiments, the sustained loss of neuromuscular function is a result of a neurotoxin such as a-bungarotoxin, a-cobratoxin, notexin, or taipoxin.

[0094] Quinolones, such as fluoroquinolone, are synthetic broad spectrum antibacterials that prevent bacterial DNA synthesis, e.g., by disrupting enzymes such as topoisomerase IV and deoxyribonucleic acid gyrase (Andersson and MacGowan, “Development of the Quinolones,” J. Antimicrob. Chemother. 51 Suppl 1 : 1-11 (2003), which is hereby incorporated by reference in its entirety). The neurotoxic effects of quinolones encompass antibiotic-associated encephalopathy, seizures, peripheral neuropathy, and exacerbation of myasthenia gravis (Anwar et al., “Fluroquinolones: Neurological Complications and Side Effects in Clinal Practice, Cureus 16(2):e54565 (2024), which is hereby incorporated by reference in its entirety). In some embodiments, the sustained loss of neuromuscular function is a result of certain antibiotics such as fluoroquinolone and natural quinoline derivatives such as quinine or quinidine.

[0095] In some embodiments, the subject has sustained loss of neuromuscular function as a result of an anti-cancer agent. In some embodiments, the sustained loss of neuromuscular function is a result of treatment with an anti-cancer agent such as vincristine, oxaliplatin, cisplatin, paclitaxel, or bortezomib (Colvin, L., “Chemotherapy-Induced Peripheral Neuropathy (CIPN): Where are we now?” Pain 160(Suppl 7 S1-S10 (2019), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has a chemotherapy induced peripheral neuropathy (CIPN) such as vincristine-induced neuropathy. In some embodiments, thesubject has a traumatic neuromuscular injury, a nerve trauma injury, a muscle trauma or injury, a contusion injury, a nerve cut, or a compression-decompression injury.

[0096] A “nerve trauma injury” refers to damage to a nerve resulting from physical injury or trauma (Campbell et al., “Evaluation and Management of Peripheral Nerve Injury,” Clinical Neurophysiology 119(9): 1951-1965 (2008), which is hereby incorporated by reference in its entirety). This type of injury can occur due to various causes such as blunt force, stretching, or crushing of the nerve. The severity of nerve trauma can range from mild, temporary dysfunction to severe, permanent damage. Symptoms may include pain, numbness, tingling, and / or muscle weakness. Treatment often involves physical therapy, medications, and in some cases, surgical intervention. In some embodiments, the subject has a nerve trauma injury.

[0097] A “nerve cut” or “neurotmesis” refers to a complete transection of a peripheral nerve (Campbell et al., “Evaluation and Management of Peripheral Nerve Injury,” Clinical Neurophysiology 119: 1951-1965 (2008), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has a nerve cut injury.

[0098] A “muscle trauma or injury” refers to any damage to muscle that results in a deficit of function (Edouard et al., “Traumatic Muscle Injury” Nature Reviews Disease Primers 9(1): 56 (2023), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has a muscle trauma or injury.

[0099] A “compression-decompression injury” refers to muscle injury refers to injury resulting from strong and / or prolonged compression of muscle such as a crush injury. Decompression injury refers to very rapid gas decompression in tissues including muscle which damages muscle cells. In some embodiments, the subject has a compression-decompression injury.

[0100] In some embodiments, the subject has a nerve trauma injury, a nerve cut, or a compression-decompression injury. In some embodiments, the subject has neuromuscular denervation or partial neuromuscular denervation.

[0101] In some embodiments, the subject has a loss of neuromuscular junctions as a result of aging, disuse atrophy, neurogenic myopathies, or a muscular dystrophy.

[0102] In some embodiments, the subject has a loss of neuromuscular junctions as a results of a neurogenic myopathy. The neurogenic myopathy may be a myopathy with neurogenic and autoimmune involvement with target fibers or tubular aggregate formation, or vacuolar myopathy.

[0103] A vacuolar myopathy is a group of muscle disorders characterized by the presence of vacuoles containing myeloid structures in muscle fibers (Mair et al., “Differential Diagnosis of Vacuolar Myopathies in the NGS Era,” Brain Pathol. 30(5):877-896 (2020), which is hereby incorporated by reference in its entirety). In some embodiments, the subject has a vascular myopathy such as bulbospinal muscular atrophy. Bulbospinal muscular atrophy is a rare, progressive motor neuron disease with predominant affection of the lower motor neuron (Finsterer and Scorza, “Central Nervous System Abnormalities in Spinal and Bulbar Muscular Atrophy (Kennedy’s Disease),” Clinical Neurology and Neurosurgery 184: 105426 (2019), which is hereby incorporated by reference in its entirety).

[0104] In some embodiments, the muscular dystrophy is spinal muscular atrophy (SMA), facioscapulohumeral muscular dystrophy (FSHD), Emery-Dreifuss muscular dystrophy (EDMD), Duchenne muscular dystrophy (DMD), limb girdle muscular dystrophy (LGMD), myotonic dystrophy, or congenital muscular dystrophy.

[0105] In some embodiments, the subject has a loss of neuromuscular junctions as a result of a muscular dystrophy and the muscular dystrophy is spinal muscular atrophy (SMA).

[0106] The administering may be effective to: (i) regenerate neuromuscular junctions in the subject; (ii) increase occurrence of neuromuscular junctions in the subject; (iii) increase occurrence of acetylcholine receptors following muscle injury in the subject; (iv) decrease occurrence of denervated myofibers following muscle injury in the subject; (v) increase occurrence of presynaptic and postsynaptic components following muscle injury in the subject; (vi) decrease acetylcholine degradation in the subject; (vii) increase mitochondrial function in motor neurons in the subject; and / or (viii) increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject.

[0107] As demonstrated in the Examples of the present disclosure, AUF1 gene therapy following muscle injury promotes expression of neuromuscular junction (NMJ) proteins and structures. Example 1 demonstrates that AUF1 gene therapy is effective to increase levels of AChR mRNA (Chrna, 0, 5,), Rapsyn mRNA, MuSK mRNA, and / or GABPA mRNA. Thus, in some embodiments, said administering is effective to increase levels of neuromuscular junction protein mRNA in the subject. In some embodiments, said administering is effective to increase levels of neuromuscular junction protein mRNA (such as AChR mRNA, Rapsyn mRNA, MuSK mRNA, and / or GABPA mRNA) by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more as compared to when said administering is not carried out. For example, administering may beeffective to increase levels of AChR mRNAs (e.g., Chrna and Chm6) by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, or any amount therebetween.

[0108] In some embodiments, administering is effective to increase levels of MuSK mRNA by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, at least 300%, or more. In some embodiments, administering is effective to increase levels of MuSK mRNA by at least 1-fold, at least 2-fold, at least 3 -fold, at least 4-fold, at least 5-fold, or any amount therebetween.

[0109] In some embodiments, administering is effective to increase levels of GABPA mRNA by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, at least 300%, or more. In some embodiments, administering is effective to increase levels of GABPA mRNA by at least 1-fold, at least 2-fold, at least 3 -fold, at least 4-fold, at least 5 -fold, or any amount therebetween.

[0110] In some embodiments, administering is effective to increase levels of Rapsyn mRNA by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, or more. In some embodiments, administering is effective to increase levels of Rapsyn mRNA by at least 1-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, or any amount therebetween.[oni] In some embodiments, said administering is effective to regenerate neuromuscular junctions in the subject. In some embodiments, said administering is effective to regenerate neuromuscular junctions in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0112] In some embodiments, said administering is effective to increase occurrence of neuromuscular junctions in the subject. In some embodiments, said administering is effective to increase occurrence of neuromuscular junctions in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0113] In some embodiments, said administering is effective to increase occurrence of acetylcholine receptors following muscle injury in the subject, e.g., as compared to when said administering is not carried out. In some embodiments, said administering is effective to increase occurrence of acetylcholine receptors following muscle injury in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0114] In some embodiments, said administering is effective to decrease occurrence of denervated myofibers following muscle injury in the subject. In some embodiments, said administering is effective to decrease occurrence of denervated myofibers following muscle injury in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out. In some embodiments, administering is effective to decrease occurrence of denervated myofibers following muscle injury in the subject by at least 1-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 4.5-fold, at least 5-fold, or any amount therebetween, e.g., as compared to when said administering is not carried out.

[0115] In some embodiments, said administering is effective to increase occurrence of presynaptic and postsynaptic components following muscle injury in the subject. In some embodiments, said administering is effective to increase occurrence of presynaptic and postsynaptic components following muscle injury in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0116] In some embodiments, said administering is effective to decrease acetylcholine degradation in the subject. In some embodiments, said administering is effective to decrease acetylcholine degradation in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0117] In some embodiments, said administering is effective to increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject. In some embodiments, said administering is effective to increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, or more, e.g., as compared to when said administering is not carried out.

[0118] Methods of measuring regeneration of neuromuscular junctions in a subject, measuring an increase in occurrence of neuromuscular junctions in a subject, measuring an increase in the occurrence of acetylcholine receptors following muscle injury in a subject, measuring a decrease in the occurrence of denervated myofibers following muscle injury in a subject, measuring an increase in the occurrence of presynaptic and postsynaptic components following muscle injury in the subject, measuring a decrease in acetylcholine degradation in the subject, and measuring an increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in a subject are well known in the art and include, without limitation, biopsy or various muscle fiber electrophysiology tests including repetitive nerve stimulation, clinical needle electromyography, and electromyographic motor unit measurements of NMJ transmission ability (Arnold and Clark, “Neuromuscular Junction Transmission Failure in Aging and Sarcopenia: The Nexus of the Neurological and Muscular Systems,” Aging Research Reviews 89: 101966 (2023), which is hereby incorporated by reference in its entirety).

[0119] In some embodiments, said administering is effective to increase in occurrence of neuromuscular junctions in a subject, increase in the occurrence of acetylcholine receptors following muscle injury in a subject, decrease the occurrence of denervated myofibers following muscle injury in a subject, increase the occurrence of presynaptic and postsynaptic components following muscle injury in the subject, and / or increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject as measured by quantification of the NMJ response or function by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 100%, or any amount therebetween. In some embodiments, said administering is effective to increase in occurrence of neuromuscular junctions in a subject, increase in the occurrence of acetylcholine receptors following muscle injury in a subject, decrease the occurrence of denervated myofibers following muscle injury in a subject, increase the occurrence of presynaptic and postsynaptic components following muscle injury in thesubject, and / or increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject as measured by quantification of the NMJ response or function to normal function.

[0120] In some embodiments, said administering is effective to increase mitochondrial function in motor neurons in the subject.

[0121] In some embodiments, said nucleic acid molecule is a vector.

[0122] The term “vector” is used interchangeably with “expression vector.” The term “vector” may refer to viral or non-viral, prokaryotic or eukaryotic, DNA or RNA sequences that are capable of being transfected into a cell, referred to as “host cell,” so that all or a part of the sequences are transcribed. It is not necessary for the transcript to be expressed. It is also not necessary for a vector to comprise a transgene having a coding sequence. Vectors are frequently assembled as composites of elements derived from different viral, bacterial, or mammalian genes. Vectors contain various coding and non-coding sequences, such as sequences coding for selectable markers, sequences that facilitate their propagation in bacteria, or one or more transcription units that are expressed only in certain cell types. For example, mammalian expression vectors often contain both prokaryotic sequences that facilitate the propagation of the vector in bacteria and one or more eukaryotic transcription units that are expressed only in eukaryotic cells. It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc.

[0123] In some embodiments, the vector is a viral vector. Suitable viral vectors are well known in the art and include, without limitation, an adeno-associated viral (AAV) vector and a lentiviral vector. In some embodiments, the viral vector is an adeno-associated viral (AAV) vector. Suitable adeno-associated viral (AAV) vectors include, without limitation, an adeno- associated virus type 8 vector or an adeno-associated virus type 9 vector. In other embodiments, the viral vector is a lentiviral vector.

[0124] Adeno-associated viral vectors and recombinant adeno-associated virus (AAV) vectors are well known delivery vehicles that can be constructed and used to deliver a nucleic acid molecule to cells, as described in Shi et al., “Therapeutic Expression of an Anti-Death Receptor-5 Single-Chain Fixed Variable Region Prevents Tumor Growth in Mice,” Cancer Res. 66: 11946-53 (2006); Fukuchi et al., “Anti-Ap Single-Chain Antibody Delivery via Adeno- Associated Virus for Treatment of Alzheimer’s Disease,” NeurobioL Dis. 23:502-511 (2006); Chatterjee et al., “Dual-Target Inhibition of HIV-1 In Vitro by Means of an Adeno-Associated Virus Antisense Vector,” Science 258: 1485-1488 (1992); Ponnazhagan et al., “Suppression ofHuman Alpha-globin Gene Expression Mediated by the Recombinant Adeno-associated Virus 2- based Antisense Vectors,” J. Exp. Med. 179:733-738 (1994), which are hereby incorporated by reference in their entirety. In vivo use of these vehicles is described in Flotte et al., “Stable In Vivo Expression of the Cystic Fibrosis Transmembrane Conductance Regulator With an Adeno- Associated Virus Vector,” Proc. Nat’l. Acad. Sci. 90: 10613-10617 (1993), which is hereby incorporated by reference in its entirety.

[0125] Recombinant adeno-associated virus (AAV) vectors provide the ability to stably transduce and express genes with very long-term (many years) duration in skeletal muscle, and depending on the AAV vector serotype and its modification, to do so with high muscle-tropism and selectivity whether using local intramuscular injection or systemic routes of delivery (Phillips et al., “Systemic Gene Transfer to Skeletal Muscle Using Reengineered AAV Vectors,” Methods Mol. Biol. 709: 141-51 (2011) and Muraine et al., “Transduction Efficiency of Adeno- Associated Virus Serotypes After Local Injection in Mouse and Human Skeletal Muscle,” Hum. Gene Ther. 31(3-4):233-240 (2020), which are hereby incorporated by reference in their entirety). Moreover, for certain AAV serotypes and engineered variants, particularly AAV8 and its engineered variants, studies in mice have been shown to be predictive of human skeletal muscle transduction and gene expression, as found in clinical trials for skeletal muscle transmission and expression (Phillips et al., “Systemic Gene Transfer to Skeletal Muscle Using Reengineered AAV V ectors,” Methods Mol. Biol. 709: 141-51 (2011) and Muraine et al., “Transduction Efficiency of Adeno- Associated Virus Serotypes After Local Injection in Mouse and Human Skeletal Muscle,” Hum. Gene Ther. 31(3-4) :233 -240 (2020), which are hereby incorporated by reference in their entirety).

[0126] The AAV vector described herein may comprise a sequence isolated or derived from an AAV vector of serotype 1 (AAV1), serotype 2 (AAV2), serotype 3 (AAV3), serotype 4 (AAV4), serotype 5 (AAV5), serotype 6 (AAV6), serotype 7 (AAV7), serotype 8 (AAV8), serotype 9 (AAV9), serotype 10 (AAV10), serotype 11 (AAV11), or any combination thereof.

[0127] In some embodiments, the adeno-associated viral (AAV) vector is a recombinant vector.

[0128] In some embodiments, the AAV vector is AAV8. AAV8 derived from macaques is very poorly immunogenic, resulting in long-term expression of the encoded transgene (for many years), and efficiently transduce skeletal muscle with high tropism and selectivity in both human and mouse (Phillips et al., “Systemic Gene Transfer to Skeletal Muscle Using Reengineered AAV V ectors,” Methods Mol. Biol. 709: 141-51 (2011); Muraine et al., “Transduction Efficiency of Adeno- Associated Virus Serotypes After Local Injection in Mouseand Human Skeletal Muscle,” Hum. Gene Ther. 31(3-4) :233 -240 (2020); Blankinship et al., “Efficient Transduction of Skeletal Muscle Using Vectors Based on Adeno-associated Virus Serotype 6,” Mol. Ther. 10(4):671-8 (2004); and Gregorevic et al., “Viral Vectors for Gene Transfer to Striated Muscle,” Curr. Opin. Mol. Ther. 6(5):491-8 (2004), which are hereby incorporated by reference in their entirety). AAV8 shows essentially no liver tropism, is largely specific for skeletal fibers and satellite cells, and has been shown to transduce skeletal muscles throughout the body (Wang et al., “Construction and Analysis of Compact Muscle-specific Promoters for AAV Vectors,” Gene Ther. 15(22): 1489-99 (2008), which is hereby incorporated by reference in its entirety).

[0129] According to some embodiments, the adeno-associated viral (AAV) vector is an AAV8 vector with the nucleotide sequence of SEQ ID NO:25.AAV8 AUF1 Construct Sequence (SEQ ID NO:25)CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGC GACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATC ACTAGGGGTTCCTGCGGCCTAAGGCAATTGGCCACTACGGGTCTAGGCTGCCCATGTAAGGAGG CAAGGCCTGGGGACACCCGAGATGCCTGGTTATAATTAACCCCAACACCTGCTGCCCCCCCCCC CCAACACCTGCTGCCTGAGCCTGAGCGGTTACCCCACCCCGGTGCCTGGGTCTTAGGCTCTGTA CACCATGGAGGAGAAGCTCGCTCTAAAAATAACCCTGTCCCTGGTGGATCGCCACTACGGGTCT AGGCTGCCCATGTAAGGAGGCAAGGCCTGGGGACACCCGAGATGCCTGGTTATAATTAACCCCA ACACCTGCTGCCCCCCCCCCCCAACACCTGCTGCCTGAGCCTGAGCGGTTACCCCACCCCGGTG CCTGGGTCTTAGGCTCTGTACACCATGGAGGAGAAGCTCGCTCTAAAAATAACCCTGTCCCTGG TGGATCGCCACTACGGGTCTAGGCTGCCCATGTAAGGAGGCAAGGCCTGGGGACACCCGAGATG CCTGGTTATAATTAACCCCAACACCTGCTGCCCCCCCCCCCCAACACCTGCTGCCTGAGCCTGA GCGGTTACCCCACCCCGGTGCCTGGGTCTTAGGCTCTGTACACCATGGAGGAGAAGCTCGCTCT AAAAATAACCCTGTCCCTGGTGGATCCCTCCCTGGGGACAGCCCCTCCTGGCTAGTCACACCCT GTAGGCTCCTCTATATAACCCAGGGGCACAGGGGCTGCCCCCGGGTCACCGCTAGCCAAAGCTT CTCGAGGCTGGCTAGTTAAGCTATCAACAAGTTTGTACAGAAAAGCAGGCTTTAAAGGAACCAA TTCAGTCGACGCTAGCAAGCTTGGTACCGGATCCGAATTCCACCATGTCGGAGGAGCAGTTCGG AGGGGACGGGGCGGCGGCGGCGGCAACGGCGGCGGTAGGCGGCTCGGCGGGCGAGCAGGAGGGA GCCATGGTGGCGGCGGCGGCGCAGGGGCCGGCGGCGGCGGCGGGAAGCGGGAGCGGCGGCGGCG GCTCTGCGGCCGGAGGCACCGAAGGAGGCAGCGCCGAGGCAGAGGGAGCCAAGATCGACGCCAG TAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACACACTGAAGCAGCGGCGGCA CAGCGGGAAGAAT GGAAAAT GT T TATAGGAGGCC T TAGC T GGGACACCACAAAGAAAGAT C T GAAGGACTACTTTTCCAAATTTGGTGAAGTTGTAGACTGCACTCTGAAGTTAGATCCTATCACAGG GCGATCAAGGGGTTTTGGCTTTGTGCTATTTAAAGAGTCGGAGAGTGTAGATAAGGTCATGGAT CAGAAAGAACATAAAT T GAAT GGGAAAGT CAT T GAT CC TAAAAGGGCCAAAGCCAT GAAAACAA AAGAG C C T G T C AAAAAAAT T T T T G T T G G T G G C C T T T C T C C AGAC AC AC C T GAAGAAAAAAT AAG AGAGTACTTTGGTGGTTTTGGTGAGGTTGAATCCATAGAGCTCCCTATGGACAACAAGACCAAT AAGAGGCGTGGGTTCTGTTTTATTACCTTTAAGGAAGAGGAGCCAGTGAAGAAGATAATGGAAA AGAAAT AC C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG C C AT G T C AAAG GAAC AG TA TCAGCAGCAGCAGCAGTGGGGATCTAGAGGAGGGTTTGCAGGCAGAGCTCGCGGAAGAGGTGGA GATCAGCAGAGTGGTTATGGGAAAGTATCCAGGCGAGGTGGACATCAAAATAGCTACAAACCAT ACTAAGATATCGCGGCCGCCTCGAGGACTACAAGGATGACGATGACAAGGATTACAAAGACGAC GATGATAAGGACTATAAGGATGATGACGACAAATAATAGCAATTCCTCGACGACTGCATAGGGT TACCCCCCTCTCCCTCCCCCCCCCCTAACGTTACTGGCCGAAGCCGCTTGGAATAAGGCCGGTG TGCGTTTGTCTATATGTTATTTTCCACCATATTGCCGTCTTTTGGCAATGTGAGGGCCCGGAAA CCTGGCCCTGTCTTCTTGACGAGCATTCCTAGGGGTCTTTCCCCTCTCGCCAAAGGAATGCAAG GTCTGTTGAATGTCGTGAAGGAAGCAGTTCCTCTGGAAGCTTCTTGAAGACAAACAACGTCTGT AGCGACCCTTTGCAGGCAGCGGAACCCCCCACCTGGCGACAGGTGCCTCTGCGGCCAAAAGCCA CGTGTATAAGATACACCTGCAAAGGCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTG TGGAAAGAGTCAAATGGCTCTCCTCAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAAGGT ACCCCATTGTATGGGATCTGATCTGGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGG TTAAAAAACGTCTAGGCCCCCCGAACCACGGGGACGTGGTTTTCCTTTGAAAAACACGATGATA ATGGCCACAACTAGTGCCACCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCA TCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGG CGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCC TGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACA TGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTT CTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTG AACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGG AGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGT GAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAG AACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCG CCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGC CGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAGTTTAAACTCTAGACCCAGCTTTCTTG TACAAAGTGGTTGATCTAGAGGGCCCGTAACTAGTTGAGCGGCCGCAACTCGAGACTCTAGAGG TTAATCGATAAT C AAC C T C T G GAT T AC AAAAT T T G T GAAAGAT T GAC TGGTATTCT T AAC T AT G TTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGG GCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGC GGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAAT TCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGA TTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCG CGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATC TCCCTTTGGGCCGCCTCCCCGCATCGAAACCCGCTGACTAGACGACTGTGCCTTCTAGTTGCCA GCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTC CTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGG GTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGATGC GGTGGGCTCTATGGCCGCGGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGC GCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGC GGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTT ACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGG CGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTA GCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAG CTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAA ACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTG ACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTA TCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGA G C T GAT T T AAC AAAAAT T T AAC G C GAAT T T T AAC AAAAT AT T AAC G T T T AC AAT TTTATGGTGC ACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCG CTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTC CGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTC GTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCA CTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTA TCCGCTCAT GAGAC AAT AAC C C T GAT AAAT G C T T C AAT AAT AT T GAAAAAG GAAGAG TAT GAG T ATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTC ACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACAT CGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATG ATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC AAC TCGGTCGCCG CAT AC AC T AT T C T C AGAAT GAC T T G G T T GAG TACT C AC C AG T C AC AGAAAA GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAAC ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAA CGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGC GAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAG GACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA GCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTT ATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTG C C T GAG T GAT T AAG C AT T G G T AAC T G T C AGAC C AAG TTTACTCATATATACTT T AGAT T GAT T T AAAAC TTCATTTTTAATT T AAAAG GAT C T AG G T GAAGAT CCTTTTTGATAATCTCAT GAG C AAA ATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTT CTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGC GGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGA GCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTG TAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA GTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGA ACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTAC AGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTAT AGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGC GGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTT TGCTCACATGT

[0130] In some embodiments, the adeno-associated virus (AAV) vector is AAV8-tMCK- AUF1 or another human AAV including but not limited to AAV1, AAV2, AAV5, AAV6, or AAV9 vector encoding AUF1 (e.g., AUF1 isoforms p37AUF1, p40AUF1, p42AUF1, and / or p45AUF1). In other embodiments, the AAV is a human novel AAV capsid variant engineered for enhanced muscle-specific tropism including but not limited to AAV2i8 or AAV2.5. In yet other embodiments, the AAV vector is a non-human primate AAV vector including but not limited to AAVrh.8, AAVrh.10, AAVrh.43, or AAVrh.74.

[0131] In some embodiments, the lentiviral vector is a lentivirus p45 AUF1 vector, or a lentivirus expressing another AUF1 isoform (e.g., p37AUF1, p40AUF1,orp42AUF1)orcombinations thereof (Abbadi et al., “Muscle Development and Regeneration Controlled by AUF1 -mediated Stage-specific Degradation of Fate-determining Checkpoint mRNAs,” Proc. Nat ’I. Acad. Sci. USA 116: 11285-90 (2019), which is hereby incorporated by reference in its entirety). Other embodiments include expression of p37AUF1, p40AUF1, p42AUF1, p45AUF1,orcombinations thereof from non-human lentivirus vectors including but not limited to simian, feline, and other mammalian lentivirus gene transfer vectors.

[0132] In some embodiments, the AUF1 p45 lentivirus vector has the following nucleotide sequence:AUF1 p45 Lentivirus Vector Shuttle Plasmid (SEQ ID NO:26)CGAAAAGTGCCACCTGCAGCCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGGGCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCCCAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGGACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCGCGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCGATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGTGGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTCATTTGGGGGCTCGTCCGGGATCGGGAGACCCCTGCCCAGGGACCACCGACCCACCACCGGGAGGCAAGCTGGCCAGCAACTTATCTGTGTCTGTCCGATTGTCTAGTGTCTATGACTGATTTTATGCGCCTGCGTCGGTACTAGTTAGCTAACTAGCTCTGTATCTGGCGGACCCGTGGTGGAACTGACGAGTTCTGAACACCCGGCCGCAACCCTGGGAGACGTCCCAGGGACTTTGGGGGCCGTTTTTGTGGCCCGACCTGAGGAAGGGAGTCGATGTGGAATCCGACCCCGTCAGGATATGTGGTTCTGGTAGGAGACGAGAACCTAAAACAGTTCCCGCCTCCGTCTGAATTTTTGCTTTCGGTTTGGAACCGAAGCCGCGCGTCTTGTCTGCTGCAGCGCTGCAGCATCGTTCTGTGTTGTCTCTGTCTGACTGTGTTTCTGTATTTGTCTGAAAATTAGGGCCAGACTGTTACCACTCCCTTAAGTTTGACCTTAGGTCACTGGAAAGATGTCGAGCGGATCGCTCACAACCAGTCGGTAGATGTCAAGAAGAGACGTTGGGTTACCTTCTGCTCTGCAGAATGGCCAACCTTTAACGTCGGATGGCCGCGAGACGGCACCTTTAACCGAGACCTCATCACCCAGGTTAAGATCAAGGTCTTTTCACCTGGCCCGCATGGACACCCAGACCAGGTCCCCTACATCGTGACCTGGGAAGCCTTGGCTTTTGACCCCCCTCCCTGGGTCAAGCCCTTTGTACACCCTAAGCCTCCGCCTCCTCTTCCTCCATCCGCCCCGTCTCTCCCCCTTGAACCTCCTCGTTCGACCCCGCCTCGATCCTCCCTTTATCCAGCCCTCACTCCTTCTCTAGGCGCCGGCCGGATCCATGTCGGAGGAGCAGTTCGGCGGGGACGGGGCGGCGGCAGCGGCAACGGCGGCGGTAGGCGGCTCGGCGGGCGAGCAGGAGGGAGCCATGGTGGCGGCGACACAGGGGGCAGCGGCGGCGGCGGGAAGCGGAGCCGGGACCGGGGGCGGAACCGCGTCTGGAGGCACCGAAGGGGGCAGCGCCGAGTCGGAGGGGGCGAAGATTGACGCCAGTAAGAACGAGGAGGATGAAGGCCATTCAAACTCCTCCCCACGACACTCTGAAGCAGCGACGGCACAGCGGG AAGAAT GGAAAAT GT T TATAGGAGGCC T TAGC T GGGACAC TACAAAGAAAGAT C T GAAGGAC TA CTTTTCCAAATTTGGTGAAGTTGTAGACTGCACTCTGAAGTTAGATCCTATCACAGGGCGATCA AGGGGTTTTGGCTTTGTGCTATTTAAAGAATCGGAGAGTGTAGATAAGGTCATGGATCAAAAAGAACATAAATTGAATGGGAAGGTGATTGATCCTAAAAGGGCCAAAGCCATGAAAACAAAAGAGCC GGTTAAAAAAATTTTTGTTGGTGGCCTTTCTCCAGATACACCTGAAGAGAAAATAAGGGAGTAC TTTGGTGGTTTTGGTGAGGTGGAATCCATAGAGCTCCCCATGGACAACAAGACCAATAAGAGGC GTGGGTTCTGCTTTATTACCTTTAAGGAAGAAGAACCAGTGAAGAAGATAATGGAAAAGAAATA C C AC AAT GTTGGTCTTAG T AAAT G T GAAAT AAAAG T AG C C AT G T C GAAG GAAC AAT AT GAG C AA CAGCAACAGTGGGGATCTAGAGGAGGATTTGCAGGAAGAGCTCGTGGAAGAGGTGGTGGCCCCA GTCAAAACTGGAACCAGGGATATAGTAACTATTGGAATCAAGGCTATGGCAACTATGGATATAA CAGCCAAGGTTACGGTGGTTATGGAGGATATGACTACACTGGTTACAACAACTACTATGGATAT GGTGATTATAGCAACCAGCAGAGTGGTTATGGGAAGGTATCCAGGCGAGGTGGTCATCAAAATA G C T AC AAAC CAT AC GAC T AC AAG GAC GAC GAT GAC AAG T GAG T C GAC C AAT TCCGGTTATTTTC CACCATATTGCCGTCTTTTGGCAATGTGAGGGCCCGGAAACCTGGCCCTGTCTTCTTGACGAGC ATTCCTAGGGGTCTTTCCCCTCTCGCCAAAGGAATGCAAGGTCTGTTGAATGTCGTGAAGGAAG CAGTTCCTCTGGAAGCTTCTTGAAGACAAACAACGTCTGTAGCGACCCTTTGCAGGCAGCGGAA CCCCCCACCTGGCGACAGGTGCCTCTGCGGCCAAAAGCCACGTGTATAAGATACACCTGCAAAG GCGGCACAACCCCAGTGCCACGTTGTGAGTTGGATAGTTGTGGAAAGAGTCAAATGGCTCTCCT CAAGCGTATTCAACAAGGGGCTGAAGGATGCCCAGAAGGTACCCCATTGTATGGGATCTGATCT GGGGCCTCGGTGCACATGCTTTACATGTGTTTAGTCGAGGTTAAAAAACGTCTAGGCCCCCCGA ACCACGGGGACGTGGTTTTCCTTTGAAAAACACGATGATAATACCATGAAAAAGCCTGAACTCA CCGCGACGTCTGTCGAGAAGTTTCTGATCGAAAAGTTCGACAGCGTCTCCGACCTGATGCAGCT CTCGGAGGGCGAAGAATCTCGTGCTTTCAGCTTCGATGTAGGAGGGCGTGGATATGTCCTGCGG GTAAATAGCTGCGCCGATGGTTTCTACAAAGATCGTTATGTTTATCGGCACTTTGCATCGGCCG CGCTCCCGATTCCGGAAGTGCTTGACATTGGGGAATTTAGCGAGAGCCTGACCTATTGCATCTC CCGCCGTGCACAGGGTGTCACGTTGCAAGACCTGCCTGAAACCGAACTGCCCGCTGTTCTGCAG CCGGTCGCGGAGGCCATGGATGCGATCGCTGCGGCCGATCTTAGCCAGACGAGCGGGTTCGGCC CATTCGGACCGCAAGGAATCGGTCAATACACTACATGGCGTGATTTCATATGCGCGATTGCTGA TCCCCATGTGTATCACTGGCAAACTGTGATGGACGACACCGTCAGTGCGTCCGTCGCGCAGGCT CTCGATGAGCTGATGCTTTGGGCCGAGGACTGCCCCGAAGTCCGGCACCTCGTGCACGCGGATT TCGGCTCCAACAATGTCCTGACGGACAATGGCCGCATAACAGCGGTCATTGACTGGAGCGAGGC GATGTTCGGGGATTCCCAATACGAGGTCGCCAACATCTTCTTCTGGAGGCCGTGGTTGGCTTGT ATGGAGCAGCAGACGCGCTACTTCGAGCGGAGGCATCCGGAGCTTGCAGGATCGCCGCGGCTCC GGGCGTATATGCTCCGCATTGGTCTTGACCAACTCTATCAGAGCTTGGTTGACGGCAATTTCGA TGATGCAGCTTGGGCGCAGGGTCGATGCGACGCAATCGTCCGATCCGGAGCCGGGACTGTCGGG CGTACACAAATCGCCCGCAGAAGCGCGGCCGTCTGGACCGATGGCTGTGTAGAAGTACTCGCCG ATAGTGGAAACCGACGCCCCAGCACTCGTCCGAGGGCAAAGGAATAGAGTAGATGCCGACCGGG ATCTATCGATAAAATAAAAGATTTTATTTAGTCTCCAGAAAAAGGGGGGAATGAAAGACCCCACCTGTAGGTTTGGCAAGCTAGCTTAAGTAACGCCATTTTGCAAGGCATGGAAAAATACATAACTG AGAATAGAGAAGT T CAGAT CAAGGT CAGGAACAGAT GGAACAGC T GAATAT GGGCCAAACAGGA TATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGAACAGCTGAATATGG GCCAAACAGGATATCTGTGGTAAGCAGTTCCTGCCCCGGCTCAGGGCCAAGAACAGATGGTCCC CAGATGCGGTCCAGCCCTCAGCAGTTTCTAGAGAACCATCAGATGTTTCCAGGGTGCCCCAAGG ACCTGAAATGACCCTGTGCCTTATTTGAACTAACCAATCAGTTCGCTTCTCGCTTCTGTTCGCG CGCTTCTGCTCCCCGAGCTCAATAAAAGAGCCCACAACCCCTCACTCGGGGCGCCAGTCCTCCG ATTGACTGAGTCGCCCGGGTACCCGTGTATCCAATAAACCCTCTTGCAGTTGCATCCGACTTGT GGTCTCGCTGTTCCTTGGGAGGGTCTCCTCTGAGTGATTGACTACCCGTCAGCGGGGGTCTTTC ACAT GCAGCAT GTAT CAAAAT TAAT TTGGTTTTTTTTCT TAAGTAT T TACAT TAAAT GGCCATA GTTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCT TCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAA AGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGG CCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCC CCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAA GATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTAC CGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGG TATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGC CCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATC GCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAG TTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGC TGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGAT CCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGG TCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTGCGGCCGCAAATCAATCTAAAGTA TATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGAT CTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAG GGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATT TATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGC CTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTG CGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCAT TCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGT TAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTT ATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTG AGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCT TCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTG CACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAG GCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTT T T T CAATAT TAT T GAAGCAT T TAT CAGGGT TAT T GT C T CAT GAG CG GAT AC AT AT T T GAAT GTA T T TAGAAAAATAAACAAATAGGGGT TCCGCGCACAT T TCCC

[0133] The term “promoter” is used interchangeably with “promoter element” and “promoter sequence.” Likewise, the term “enhancer” is used interchangeably with “enhancer element” and “enhancer sequence.” The term “promoter” refers to a minimal sequence of a transgene that is sufficient to initiate transcription of a coding sequence of the transgene. Promoters may be constitutive or inducible. A constitutive promoter is considered to be a strong promoter if it drives expression of a transgene at a level comparable to that of the cytomegalovirus promoter (CMV) (Boshart et al., “A Very Strong Enhancer is Located Upstream of an Immediate Early Gene of Human Cytomegalovirus,” Cell 41 :521 (1985), which is hereby incorporated by reference in its entirety). Promoters may be synthetic, modified, or hybrid promoters. Promoters may be coupled with other regulatory sequences / elements which, when bound to appropriate intracellular regulatory factors, enhance (“enhancers”) or repress (“repressors”) promoter-dependent transcription. A promoter, enhancer, or repressor, is said to be “operably linked” to a transgene when such element(s) control(s) or affect(s) transgene transcription rate or efficiency. For example, a promoter sequence located proximally to the 5' end of a transgene coding sequence is usually operably linked with the transgene. As used herein, the term “regulatory elements” is used interchangeably with “regulatory sequences” and refers to promoters, enhancers, and other expression control elements, or any combination of such elements.

[0134] Promoters are positioned 5' (upstream) to the genes that they control. Many eukaryotic promoters contain two types of recognition sequences: TATA box and the upstream promoter elements. The TATA box, located 25-30 bp upstream of the transcription initiation site, is thought to be involved in directing RNA polymerase II to begin RNA synthesis at the correct site. In contrast, the upstream promoter elements determine the rate at which transcription is initiated. These elements can act regardless of their orientation, but they must be located within 100 to 200 bp upstream of the TATA box.

[0135] Enhancer elements can stimulate transcription up to 1000-fold from linked homologous or heterologous promoters. Enhancer elements often remain active even if their orientation is reversed (Li et al., “High Level Desmin Expression Depends on a Muscle-SpecificEnhancer,” J. Bio. Chem. 266(10):6562-6570 (1991), which is hereby incorporated by reference in its entirety). Furthermore, unlike promoter elements, enhancers can be active when placed downstream from the transcription initiation site, e.g., within an intron, or even at a considerable distance from the promoter (Yutzey et al., “An Internal Regulatory Element Controls Troponin I Gene Expression,” Mol. Cell. Bio. 9(4): 1397-1405 (1989), which is hereby incorporated by reference in its entirety).

[0136] The term “muscle cell-specific” refers to the capability of regulatory elements, such as promoters and enhancers, to drive expression of an operatively linked nucleic acid molecule (e.g., a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof) exclusively or preferentially in muscle cells or muscle tissue.

[0137] The nucleic acid molecules disclosed herein may comprise a muscle cell-specific promoter. In some embodiments, the muscle cell-specific promoter mediates cell-specific and / or tissue-specific expression of an AUF1 protein or fragment thereof. The promoter may be a mammalian promoter. For example, the promoter may be a human promoter, a murine promoter, a porcine promoter, a feline promoter, a canine promoter, an ovine promoter, a nonhuman primate promoter, an equine promoter, a bovine promoter, or the like.

[0138] In some embodiments, the muscle cell-specific promoter is a naturally occurring muscle cell-specific promoter. For example, the muscle cell-specific promoter may be aa muscle creatine kinase (MCK) promoter, a smooth muscle 22 (SM22) promoter, a myo-3 promoter, a creatine kinase (CK) 8 promoter, a U6 promoter, a Hl promoter, a Desmin promoter, a Pitx3 promoter, or a skeletal alpha-actin promoter.

[0139] In some embodiments, the muscle cell-specific promoter is a synthetic muscle cell- specific promoter. For example, the muscle cell specific promoter may be a C5-12 promoter, a CK6-CK9 promoter, a dMCK promoter, a tMCK promoter, a smooth muscle 22 (SM22) promoter, a myo-3 promoter, a Spc512 promoter, a creatine kinase (CK) 8 promoter, a creatine kinase (CK) 8e promoter, a U6 promoter, a Hl promoter, a desmin promoter, a Pitx3 promoter, a skeletal alpha-actin promoter, a MHCK7 promoter, or a Sp-301 promoter.

[0140] Suitable muscle cell-specific promoter sequences are well known in the art and are provided in Table 2 below (Malerba et al., “PABPN1 Gene Therapy for Oculopharyngeal Muscular Dystrophy,” Nat. Commun. 8: 14848 (2017); Wang et al., “Construction and Analysis of Compact Muscle-Specific Promoters for AAV Vectors,” Gene. Ther. 15: 1489-1499 (2008); Piekarowicz et al., “A Muscle Hybrid Promoter as a Novel Tool for Gene Therapy,” Mol. Ther. Methods Clin. Dev. 15: 157-169 (2019); Salva et al., “Design of Tissue-Specific RegulatoryCassettes for High-Level rAAV-Mediated Expression in Skeletal and Cardiac Muscle,” Mol. Ther. 15(2):320-329 (2007); Lui et al., “Synthetic Promoter for Efficient and Muscle-Specific Expression of Exogenous Genes,” Plasmid 106: 102441 (2019), which are hereby incorporated by reference in their entirety).Table 2: Muscle Specific-Promoter Sequences*See Malerba et al., “PABPN1 Gene Therapy for Oculopharyngeal Muscular Dystrophy,” Nat. Commun. 8: 14848 (2017); Wang et al., “Construction and Analysis of Compact Muscle-Specific Promoters for AAV Vectors,” Gene. Ther. 15: 1489-1499 (2008); Piekarowicz et al., “A Muscle Hybrid Promoter as a Novel Tool for Gene Therapy,” Mol. Ther. Methods Clin. Dev. 15: 157-169 (2019); and Salva et al., “Design of Tissue-Specific Regulatory Cassettes for High-Level rAAV- Mediated Expression in Skeletal and Cardiac Muscle,” Mol. Ther. 15(2):320-329 (2007), which are hereby incorporated by reference in their entirety.

[0141] In some embodiments, the muscle cell-specific promoter is a muscle creatinekinase (“MCK”) promoter. The muscle creatine kinase (MCK) gene is highly active in all striated muscles. Creatine kinase plays an important role in the regeneration of ATP within contractile and ion transport systems. It allows for muscle contraction when neither glycolysis nor respiration is present by transferring a phosphate group from phosphocreatine to ADP to form ATP. There are four known isoforms of creatine kinase: brain creatine kinase (CKB), muscle creatine kinase (MCK), and two mitochondrial forms (CKMi). MCK is the most abundant non-mitochondrial mRNA that is expressed in all skeletal muscle fiber types and is also highly active in cardiac muscle. The MCK gene is not expressed in myoblasts, but becomes transcriptionally active when myoblasts commit to terminal differentiation into myocytes. MCK gene regulatory regions display striated muscle-specific activity and have been extensively characterized in vivo and in vitro. The major known regulatory regions in the MCK gene include a muscle-specific enhancer located approximately 1.1 kb 5' of the transcriptional start site in mouse and a 358-bp proximal promoter. Additional sequences that modulate MCK expression are distributed over 3.3 kb region 5' of the transcriptional start site and in the 3.3-kb first intron. Mammalian MCK regulatory elements, including human and mouse promoter and enhancer elements, are described in Hauser et al., “Analysis of Muscle Creatine Kinase RegulatoryElements in Recombinant Adenoviral Vectors,” Mol. Therapy 2: 16-25 (2000), which is hereby incorporated by reference in its entirety. Suitable muscle creatine kinase (MCK) promoters include, without limitation, a wild type MCK promoter, a dMCK promoter, and a tMCK promoter (Wang et al., “Construction and Analysis of Compact Muscle-Specific Promoters for AAV Vectors,” Gene Ther. 15(22): 1489-1499 (2008), which is hereby incorporated by reference in its entirety).

[0142] In some embodiments of the methods disclosed herein, the nucleic acid molecule encoding the AUF1 protein or functional fragment thereof is heterologous to and operatively coupled to a muscle cell-specific promoter (e.g., a muscle creatine-kinase (tMCK) promoter).

[0143] In some embodiments, the composition comprises a lipid nanoparticle.

[0144] In some embodiments, the lipid nanoparticle is a muscle-tropic lipid nanoparticle.

[0145] In some embodiments, the lipid nanoparticle is a nerve-tropic lipid nanoparticle.

[0146] The methods and compositions described herein may be used in combination with other known treatments or standards of care for given diseases, injury, or conditions. For example, in some embodiments, the method further involves administering a drug or gene therapy effective to treat a neuromuscular disease or disorder in the subject.

[0147] In some embodiments of the methods according to the present disclosure, where the method involve administering a drug or gene therapy effective to treat a neuromuscular disease or disorder in the subject, the: (i) the drug or gene therapy may comprise an adeno- associated virus 9 vector-based gene therapy; (ii) the drug or gene therapy may comprise an antisense oligonucleotide, an siRNA, an shRNA, or miRNA; and / or (iii) the subject may have spinal muscular atrophy (SMA) or amyotrophic lateral sclerosis (ALS).

[0148] In some embodiments, the drug or gene therapy comprises an adeno-associated virus 9 vector-based gene therapy.

[0149] In some embodiments, the drug or gene therapy comprises an antisense oligonucleotide, an siRNA, an shRNA, or miRNA. siRNAs are double stranded synthetic RNA molecules approximately 20-25 nucleotides in length with short 2-3 nucleotide 3' overhangs on both ends. The double stranded siRNA molecule represents the sense and anti-sense strand of a portion of the target mRNA molecule.

[0150] siRNA molecules are typically designed to target a region of the mRNA target approximately 50-100 nucleotides downstream from the start codon. Methods and online tools for designing suitable siRNA sequences based on the target mRNA sequences are readily available in the art see, e.g., Reynolds et al., “Rational siRNA Design for RNA Interference,” Nat. Biotech. 2:326-330 (2004); Chalk et al., “Improved and Automated Prediction of EffectivesiRNA,” Biochem. Biophys. Res. Comm. 319(l):264-274 (2004); Zhang et al., “Weak Base Pairing in Both Seed and 3' Regions Reduces RNAi Off-targets and Enhances si / shRNA Designs,” Nucleic Acids Res. 42(19): 12169-76 (2014), which are hereby incorporated by reference in their entirety). Upon introduction into a cell, the siRNA complex triggers the endogenous RNA interference (RNAi) pathway, resulting in the cleavage and degradation of the target mRNA molecule.

[0151] Short or small hairpin RNA (“shRNA”) molecules are similar to siRNA molecules in function, but comprise longer RNA sequences that make a tight hairpin turn. shRNA is cleaved by cellular machinery into siRNA and gene expression is silenced via the cellular RNA interference pathway. Methods and tools for designing suitable shRNA sequences based on the target mRNA sequences are readily available in the art (see e.g., Taxman et al., “Criteria for Effective Design, Constructions, and Gene Knockdown shRNA Vectors,” BMC Biotech. 6:7 (2006) and Taxman et al., “Short Hairpin RNA (shRNA): Design, Delivery, and Assessment of Gene Knockdown,” Meth. Mol. Biol. 629: 139-156 (2010), which are hereby incorporated by reference in their entirety).

[0152] Other suitable agents that can be encoded by the recombinant construct disclosed herein include microRNAs (“miRNAs”). miRNAs are small, regulatory, noncoding RNA molecules that control the expression of their target mRNAs predominantly by binding to the 3' untranslated region (UTR). A single UTR may have binding sites for many miRNAs or multiple sites for a single miRNA, suggesting a complex post-transcriptional control of gene expression exerted by these regulatory RNAs (Shulka et al., “MicroRNAs: Processing, Maturation, Target Recognition and Regulatory Functions,” Mol. Cell. Pharmacol. 3(3):83-92 (2011), which is hereby incorporated by reference in its entirety). Mature miRNA are initially expressed as primary transcripts known as a pri-miRNAs which are processed, in the cell nucleus, to 70- nucleotide stem-loop structures called pre-miRNAs by the microprocessor complex. The dsRNA portion of the pre-miRNA is bound and cleaved by Dicer to produce a mature 22 bp double-stranded miRNA molecule that can be integrated into the RISC complex; thus, miRNA and siRNA share the same cellular machinery downstream of their initial processing.

[0153] In some embodiments, the subject has spinal muscular atrophy (SMA) and the gene therapy encodes an SMN1 gene. In some embodiments, the gene therapy is ZOLGENSMA® (onasemnogene abeparvovec-xioi), nusinersen (Spinraza), or risdiplam (Evrysdi).

[0154] In some embodiments, the subject has Duchenne muscular dystrophy (DMD) and a mutation in the DMD gene, wherein said mutation is not in exon 8 and / or exon 9 of DMDgene; and the gene therapy encodes a micro-dystrophin. In accordance with such embodiments, the gene therapy may involves exon skipping approaches for the dystrophin mRNA.

[0155] In some embodiments, the gene therapy is ELEVIDYS micro-dystrophin (delandistrogene moxeparvovec-rokl).

[0156] In some embodiments, the subject has amyotrophic lateral sclerosis (ALS). In accordance with such embodiments, the subject may have ALS associated with a mutation in the superoxide dismutase 1 (SOD1) gene and the drug comprises an antisense oligonucleotide targeting SOD1 mRNA. In some embodiments, the drug is tofersen (Qalsody).

[0157] In some embodiments, the drug is a benzothiazole. For example, the drug may be RILUTEX® (riluzole), EXSERVAN™ (riluzole), or TIGLUTIK® (riluzole).

[0158] In some embodiments, the composition and the drug or gene therapy are administered simultaneously.

[0159] In some embodiments, the composition and the drug or gene therapy are administered sequentially.

[0160] In some embodiments, said administering the composition and / or administering the drug or gene therapy are carried out by systemic or local administration.

[0161] In some embodiments, said administering the composition and / or administering the drug or gene therapy is carried out by intramuscular injection.

[0162] As described in more detail below, compositions according to the present application may be useful in gene therapy. Delivery of the active agent of a composition described herein in vivo may involve a process that effectively introduces a molecule of interest (e.g., AUF1 protein or a functional fragment thereof) into the cells or tissue being treated. In the case of polypeptide-based active agents, this can be carried out directly or, alternatively, by transfecting transcriptionally active DNA into living cells such that the active polypeptide coding sequence is expressed and the polypeptide is produced by cellular machinery. Transcriptionally active DNA may be delivered into the cells or tissue, e.g., muscle, being treated using transfection methods including, but not limited to, electroporation, microinjection, calcium phosphate coprecipitation, DEAE dextran facilitated transfection, cationic liposomes, and retroviruses. In certain embodiments, the DNA to be transfected is cloned into a vector.

[0163] Alternatively, cells can be engineered in vivo by administration of the polynucleotide using techniques known in the art. For example, by direct injection of a “naked” polynucleotide (Feigner et al., “Gene Therapeutics,” Nature 349:351-352 (1991); U.S. Patent No. 5,679,647; Wolff et al., “The Mechanism of Naked DNA Uptake and Expression,” Adv. Genet. 54:3-20 (2005), which are hereby incorporated by reference in their entirety) or apolynucleotide formulated in a composition with one or more other targeting elements which facilitate uptake of the polynucleotide by a cell.

[0164] Also encompassed are expression systems comprising nucleic acid molecules described herein. Generally, the use of recombinant expression systems involves inserting a nucleic acid molecule encoding the amino acid sequence of a desired peptide into an expression system to which the molecule is heterologous (z.e., not native or not normally present). One or more desired nucleic acid molecules encoding a peptide described herein (e.g., AUF1) may be inserted into the vector. When multiple nucleic acid molecules are inserted, the multiple nucleic acid molecules may encode the same or different peptides. The heterologous nucleic acid molecule is inserted into the expression system or vector in proper sense (5'^3') orientation relative to the promoter and any other 5' regulatory molecules, and correct reading frame.

[0165] The preparation of the nucleic acid constructs can be carried out using standard cloning procedures well known in the art as described by Joseph Sambrook et al., MOLECULAR CLONING: A LABORATORY ANUAL (Cold Springs Harbor 2012), which is hereby incorporated by reference in its entirety. U.S. Patent No. 4,237,224 to Cohen and Boyer, which is hereby incorporated by reference in its entirety, describes the production of expression systems in the form of recombinant plasmids using restriction enzyme cleavage and ligation with DNA ligase. These recombinant plasmids are then introduced by means of transformation and replicated in a suitable host cell.

[0166] A nucleic acid molecule encoding an AUF1 protein or functional fragment thereof and that is operatively coupled to a muscle-cell specific promoter (e.g., muscle creatine kinase (MCK) promoter) may include an additional elements including, without limitation, a leader sequence, a suitable 3' regulatory region to allow transcription in the host or a certain medium, and / or any additional desired component, such as reporter or marker genes. Such additional elements may be cloned into the vector of choice using standard cloning procedures in the art, such as described in Joseph Sambrook et al., MOLECULAR CLONING: A LABORATORY MANUAL (Cold Springs Harbor 2012); Frederick M. Ausubel, SHORT PROTOCOLS IN MOLECULAR BIOLOGY (Wiley 2002); and U.S. Patent No. 4,237,224 to Cohen and Boyer, which are hereby incorporated by reference in their entirety.

[0167] There are other specific initiation signals required for efficient gene transcription and translation in eukaryotic cells that can be included in the nucleic acid construct to maximize protein production. Depending on the vector system and host utilized, any number of suitable transcription and / or translation elements, including constitutive, inducible, and repressible promoters, as well as minimal 5' promoter elements, enhancers or leader sequences may be used.

[0168] In some embodiments, the composition of the present disclosure further comprises a buffer solution.

[0169] The composition of the present application may further comprise one or more targeting elements. Suitable targeting elements include, without limitation, agents such as saponins or cationic polyamides (see, e.g., U.S. Patent Nos. 5,739,118 and 5,837,533, which are hereby incorporated by reference in their entirety); microparticles, microcapsules, liposomes, or other vesicles; lipids; cell-surface receptors; transfecting agents; peptides (e.g., one known to enter the nucleus); or ligands (such as one subject to receptor-mediated endocytosis). Suitable means for using such targeting elements include, without limitation: microparticle bombardment; coating the polynucleotide with lipids, cell-surface receptors, or transfecting agents; encapsulation of the polynucleotide in liposomes, microparticles, or microcapsules; administration of the polynucleotide linked to a peptide which is known to enter the nucleus; or administration of the polynucleotide linked to a ligand subject to receptor-mediated endocytosis (see, e.g., Wu et al., “Receptor-Mediated in vitro Gene Transformation by a Soluble DNA Carrier System,” J. Biol. Chem. 262:4429-4432 (1987), which is hereby incorporated by reference in its entirety), which can be used to target cell types specifically expressing the receptors. Alternatively, a polynucleotide-ligand complex can be formed allowing the polynucleotide to be targeted for cell specific uptake and expression in vivo by targeting a specific receptor (see, e.g., PCT Application Publication Nos. WO 92 / 06180, WO 92 / 22635, WO 92 / 203167, WO 93 / 14188, and WO 93 / 20221, which are hereby incorporated by reference in their entirety).

[0170] In some embodiments, the composition is an aqueous composition. Aqueous compositions of the present application comprise an effective amount of the vector, dissolved or dispersed in a pharmaceutically acceptable carrier or aqueous medium.

[0171] In some embodiments, the composition is a pharmaceutical composition comprising a nucleic acid molecule or vector according to the present disclosure and a pharmaceutically-acceptable carrier.

[0172] The term “pharmaceutically acceptable carrier” refers to a carrier that does not cause an allergic reaction or other untoward effect in patients to whom it is administered and are compatible with the other ingredients in the formulation. Pharmaceutically acceptable carriers include, for example, pharmaceutical diluents, excipients, or carriers suitably selected with respect to the intended form of administration, and consistent with conventional pharmaceutical practices. For example, solid carriers / diluents include, but are not limited to, a gum, a starch (e.g., com starch, pregelatinized starch), a sugar (e.g., lactose, mannitol, sucrose, dextrose), acellulosic material (e.g, microcrystalline cellulose), an acrylate (e.g., polymethylacrylate), calcium carbonate, magnesium oxide, talc, or mixtures thereof. Pharmaceutically acceptable carriers may further comprise minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the nucleic acid molecule described herein.

[0173] The vector(s) (z.e., adeno-associated viral (AAV) vector and / or lentiviral vectors disclosed herein) and / or pharmaceutical composition(s) disclosed herein can be formulated according to any available conventional method. Examples of preferred dosage forms include a tablet, a powder, a subtle granule, a granule, a coated tablet, a capsule, a syrup, a troche, an inhalant, a suppository, an injectable, an ointment, an ophthalmic ointment, an eye drop, a nasal drop, an ear drop, a cataplasm, a lotion and the like. In the formulation, generally used additives such as a diluent, a binder, a disintegrant, a lubricant, a colorant, a flavoring agent, and if necessary, a stabilizer, an emulsifier, an absorption enhancer, a surfactant, a pH adjuster, an antiseptic, an antioxidant, and the like can be used.

[0174] In addition, formulating a pharmaceutical composition can be carried out by combining compositions that are generally used as a raw material for pharmaceutical formulation, according to conventional methods. Examples of these compositions include, for example, (1) an oil such as a soybean oil, a beef tallow and synthetic glyceride; (2) hydrocarbon such as liquid paraffin, squalene, and solid paraffin; (3) ester oil such as octyldodecyl myristic acid and isopropyl myristic acid; (4) higher alcohol such as cetostearyl alcohol and behenyl alcohol; (5) a silicon resin; (6) a silicon oil; (7) a surfactant such as polyoxyethylene fatty acid ester, sorbitan fatty acid ester, glycerin fatty acid ester, polyoxyethylene sorbitan fatty acid ester, a solid polyoxyethylene castor oil and polyoxyethylene polyoxypropylene block co-polymer; (8) water soluble macromolecule such as hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl polymer, polyethyleneglycol, polyvinylpyrrolidone and methylcellulose; (9) lower alcohol such as ethanol and isopropanol; (10) multivalent alcohol such as glycerin, propyleneglycol, dipropyleneglycol and sorbitol; (11) a sugar such as glucose and cane sugar; (12) an inorganic powder such as anhydrous silicic acid, aluminum magnesium silicicate, and aluminum silicate; (13) purified water, and the like.

[0175] Additives for use in the above formulations may include, for example, (1) lactose, com starch, sucrose, glucose, mannitol, sorbitol, crystalline cellulose, and silicon dioxide as the diluent; (2) polyvinyl alcohol, polyvinyl ether, methyl cellulose, ethyl cellulose, gum arabic, tragacanth, gelatine, shellac, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, polyvinylpyrrolidone, polypropylene glycol-poly oxyethylene-block co-polymer, meglumine,calcium citrate, dextrin, pectin, and the like as the binder; (3) starch, agar, gelatine powder, crystalline cellulose, calcium carbonate, sodium bicarbonate, calcium citrate, dextrin, pectic, carboxymethylcellulose / calcium, and the like as the disintegrant; (4) magnesium stearate, talc, polyethyleneglycol, silica, condensed plant oil, and the like as the lubricant; (5) any colorant whose addition is pharmaceutically acceptable is adequate as the colorant; (6) cocoa powder, menthol, aromatizer, peppermint oil, cinnamon powder as the flavoring agent; (7) antioxidants whose addition is pharmaceutically accepted such as ascorbic acid or alpha-tophenol.

[0176] Administering, according to the present disclosure, may be carried out by oral administration, topical administration, transdermal administration, parenteral administration, subcutaneous administration, intravenous administration, intramuscular administration, intraperitoneal administration, by intranasal instillation administration, by intracavitary or intravesical instillation, intraocular administration, intraarterial administration, intralesional administration, or by application to mucous membranes. Thus, in some embodiments, the administering is carried out by intramuscular administration, intravenous administration, subcutaneous administration, oral administration, or intraperitoneal administration to a subject. In specific embodiments, the administering is carried out by intramuscular injection.

[0177] In some embodiments, the administering is effective to deliver the nucleic acid molecule described herein to a specific tissue in the subject. The tissue may be muscle tissue. For example, the muscle tissue may be all types of skeletal muscle, smooth muscle, or cardiac muscle.

[0178] Formulations for injection may be presented in unit dosage form, e.g., in ampoules or in multi -dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and / or dispersing agents.

[0179] Suitable regimens for initial contacting and further doses or for sequential contacting steps may all be the same or may be variable. Appropriate regimens can be ascertained by the skilled artisan, from the disclosure of the present application, the documents cited herein, and the knowledge in the art.

[0180] A dosage unit to be administered in methods of the present application will vary depending on the vector used, the route of administration, the type of tissue and cell being targeted, and the purpose of treatment, among other parameters. Dosage for treatment can be determined by a skilled person who would know how to determine dose using methods standard in the art. A dosage unit, corresponding to genome copy number, for example, could range from about, IxlO1to IxlO11, IxlO2to IxlO11, IxlO3to IxlO11, IxlO4to IxlO11, IxlO5to IxlO11, IxlO6to IxlO11, IxlO7to IxlO11, IxlO8to IxlO11, IxlO9to IxlO11, IxlO10to IxlO11, IxlO1to IxlO10, IxlO2to IxlO10, IxlO3to IxlO10, IxlO4to IxlO10, IxlO5to IxlO10, IxlO6to IxlO10, IxlO7to IxlO10, IxlO8to IxlO10, IxlO9to IxlO10, IxlO1to IxlO9, IxlO2to IxlO9, IxlO3to IxlO9, IxlO4to IxlO9, IxlO5to IxlO9, IxlO6to IxlO9, IxlO7to IxlO9, IxlO8to IxlO9, IxlO1to IxlO8, IxlO2to IxlO8, IxlO3to IxlO8, IxlO4to IxlO8, IxlO5to IxlO8, IxlO6to IxlO8, or IxlO7to IxlO8genome copies of a vector disclosed herein. In some embodiments, a dosage unit, corresponding to genome copy number, for example, is administered in the range of IxlO1to IxlO12, IxlO2to IxlO12, IxlO3to IxlO12, IxlO4to IxlO12, IxlO5to IxlO12, IxlO6to IxlO12, IxlO7to IxlO12, IxlO8to IxlO12, IxlO9to IxlO12, IxlO10to IxlO12, or IxlO11to IxlO12genome copies; IxlO1to IxlO13, IxlO2to IxlO13, IxlO3to IxlO13, IxlO4to IxlO13, IxlO5to IxlO13, IxlO6to IxlO13, IxlO7to IxlO13, IxlO8to IxlO13, IxlO9to IxlO13, IxlO10to IxlO13, IxlO11to IxlO13, or IxlO12to IxlO13genome copies; IxlO1to IxlO14, IxlO2to IxlO14, IxlO3to IxlO14, IxlO4to IxlO14, IxlO5to IxlO14, IxlO6to IxlO14, IxlO7to IxlO14, IxlO8to IxlO14, IxlO9to IxlO14, IxlO10to IxlO14, IxlO11to IxlO14, IxlO12to IxlO14, or IxlO13to IxlO14genome copies; IxlO1to IxlO15, IxlO2to IxlO15, IxlO3to IxlO15, IxlO4to IxlO15, IxlO5to IxlO15, IxlO6to IxlO15, IxlO7to IxlO15, IxlO8to IxlO15, IxlO9to IxlO5, IxlO10to IxlO15, IxlO11to IxlO15, IxlO12to IxlO15, IxlO13to IxlO15, or IxlO14to IxlO15genome copies; IxlO1to IxlO16, IxlO2to IxlO16, IxlO3to IxlO16, IxlO4to IxlO16, IxlO5to IxlO16, IxlO6to IxlO16, IxlO7to IxlO16, IxlO8to IxlO16, IxlO9to IxlO16, IxlO10to IxlO16, IxlO11to IxlO16, IxlO12to IxlO16, IxlO13to IxlO16, IxlO14to IxlO16, or IxlO15to IxlO16genome copies; IxlO1to 3xl016, IxlO2to 3xl016, IxlO3to 3xl016, IxlO4to 3xl016, IxlO5to 3xl016, IxlO6to 3xl016, IxlO7to 3xl016, IxlO8to 3xl016, IxlO9to 3xl016, IxlO10to 3xl016, IxlO11to 3xl016, IxlO12to 3xl016, IxlO13to 3xl016, IxlO14to 3xl016, or IxlO15to 3xl016genome copies; and any amount there between. Dosage will depend on route of administration, type of tissue and cells to receive the vector, timing of administration to human subjects, whether dosage is determined based on total genome copies to be delivered, and whether administration is determined by genome copies per kilogram body weight.

[0181] In some embodiments, a subject is administered the composition described herein in one dose. In other embodiments, the subject is administered the composition described herein in a series of two or more doses in succession. In some other embodiments, where the subject is administered the composition described herein in a single dose, in two doses, and / or more than two doses, the doses may be the same or different, and they are administered with equal or with unequal intervals between them.

[0182] A subject may be administered the composition described herein in many frequencies over a wide range of times. In some embodiments, the subject is administered thevector or pharmaceutical composition described herein over a period of less than one day. In other embodiments, the subject is contacted over two, three, four, five, or six days. In some embodiments, the contacting is carried out one or more times per week, over a period of weeks. In other embodiments, the contacting is carried out over a period of weeks for one to several months. In various embodiments, the contacting is carried out over a period of months. In others, the contacting may be carried out over a period of one or more years. Generally, lengths of treatment will be proportional to the length of the ischemic disease process, the effectiveness of the therapies being applied, and the condition and response of the subject being treated. According to some embodiments, the contacting is carried out daily.

[0183] The choice of formulation for administering the composition described herein will depend on a variety of factors. Prominent among these will be the species of subject, the nature of the disorder, dysfunction, or disease being treated and its state and distribution in the subject, the nature of other therapies and agents that are being administered, the optimum route for administration, survivability via the route, the dosing regimen, and other factors that will be apparent to those skilled in the art. In particular, for instance, the choice of suitable carriers and other additives will depend on the exact route of contacting and the nature of the particular dosage form.

[0184] In the methods described herein, rather than administering a nucleic acid molecule or vector according to the present disclosure, other means of administering AUF can be carried out including by direct injection of: (i) encoding p37AUF1, p40AUF1, p42AUF1, and / or p45AUF1DNA by plasmid; (ii) mRNA encoding p37AUF1, p40AUF1, p42AUF1, and / or p45AUF1; and / or (iii) nanoparticle incorporation of AUF1 encoding DNA or mRNA.EXAMPLES

[0185] The examples below are intended to exemplify the practice of embodiments of the present application but are by no means intended to limit the scope thereof.Materials and Methods for Example 1MiceAll animal studies were approved by the NYU School of Medicine Institutional Animal Care and Use Committee (IACUC) and conducted in accordance with IACUC guidelines. 2 month or 3 month old C57BL6 mice (Jackson) for AUF1 supplementation during AAV experiments.Cells

[0186] C2C12 cells were obtained from the American Type Culture Collection (ATCC), authenticated by STR profiling and routinely checked for mycoplasma contamination. C2C12 cells were maintained in DMEM (Coming), 10% FBS (Gibco), and 1% penicillin streptomycin (Life Technologies). To differentiate cells, media was switched to DMEM (Coming), 2% Horse Serum (Gibco), and 1% penicillin streptomycin (Life Technologies) during 96 hours (Panda et al., “RNA-Binding Protein AUF1 Promotes Myogenesis by Regulating MEF2C Expression Levels,” Mol. Cell Biol. 34(16): 3106-3119 (2014), which is hereby incorporated by reference in its entirety), aufl KO C2C12 cells were created with Crispr-Cas9 methods (Abbadi et al., “Muscle Development and Regeneration Controlled by AUF1 -Mediated Stage-Specific Degradation of Fate-Determining Checkpoint mRNAs,” Proc. Natl. Acad. Sci. USA 116(23): 11285-11290 (2019), which is hereby incorporated by refemce in its entirety). For assays performed in the presence of actinomycin D to determine mRNA stability, C2C12 myoblasts cells were treated with 0.2 pg / ml of actinomycin D (Sigma). RNA immune- precipitation experiments were done in WT C2C12 before and 48 hours of differentiation using a normal IgG rabbit control or a rabbit-anti AUF1 antibody (07-260, Millipore).Immunofluorescence

[0187] Mice had skeletal muscles removed as indicated in the text, put in OCT, frozen in dry ice-cooled isopentane (Tissue-Tek), fixed in 4% paraformaldehyde, and blocked in 3% BSA in TBS. C2C12 cells were fixed in 4% paraformaldehyde and blocked in 3% BSA in PBS. Samples were immunostained overnight with antibodies. Alexa Fluor donkey 488 and 555 secondary antibodies were used at 1 :300 and incubated for 1 hour at room temperature. Slides were sealed with Vectashield with DAPI (Vector). Images were processed using ImageJ.Microscopy, Image Processing, and Analysis

[0188] Images were acquired using a Zeiss LSM 700 confocal microscope, primarily with the 20X lens. Images were processed using ImageJ. If needed, color balance was adjusted linearly for the entire image and all images in experimental sets.Real-Time PCR Analysis

[0189] RNA was extracted using Trizol (Invitrogen) according to the manufacturer’s instructions. DNase treatment was systematically performed. Quantification of extracted RNA was assessed using Nanodrop. The cDNA was synthesized using High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). mRNA was analyzed by real-time PCR using the iTaq Universal SYBR Green Supermix (Bio Rad) probe. Relative quantification was determinedusing the comparative CT method with data normalized to housekeeping gene and calibrated to the average of control groups.AA V-A UF1 Expression / AA V A UF1 Gene Transfer

[0190] AUF1 was integrated into an AAV8 vector under the tMCK promoter (AAV8- tMCK-AUFl-IRES-eGFP) (Vector Biolabs). AAV8-tMCK-IRES-eGFP was used as a control vector. This promoter was generated by the addition of a triple tandem of 2RS5 enhancer sequences (3 -Ebox) ligated to the truncated regulation region of the MCK (muscle creatine kinase) promoter, which induced high muscle specificity (Wang et al., “Construction and Analysis of Compact Muscle-Specific Promoters for AAV Vectors,” Gene Ther. 15(22): 1489- 1499 (2008), which is hereby incorporated by reference in its entirety). C57B16 mice were injected with a single retro-orbital injection of 50 pl (final concentration: 2.5xlOnparticles).Quantification and Statistical Analysis

[0191] All results are expressed as the mean ± SEM. Two group comparisons were analyzed by the unpaired Mann-Whitney test. Multiple group comparisons were performed using one-way analysis of variance (ANOVA). The non-parametric Kruskal-Wallis test followed by the Dunn’s comparison of pairs was used to analyze groups when suitable. -values of <0.05 were considered significant. All statistical analyses were performed using GraphPad Prism (version 7) software.Example 1 - AUF1 Gene Therapy Following Muscle Injury Promotes Expression of Neuromuscular Junction (NMJ) Proteins and Structures

[0192] Therapeutic promotion of muscle mass and regeneration following injury, or for aging loss, is not productive if it does not also promote NMJ generation. Pathological loss of NMJs and neuromuscular transmission is common in injured muscle, sarcopenia and atrophic muscle injury in humans and in mice (Rudolf et al., “Degeneration of Neuromuscular Junction in Age and Dystrophy,” Front. Aging Neurosci. 6:99 (2014) and Rudolf et al., “Neuromuscular Junction Degeneration in Muscle Wasting,” Curr. Opin. Clin. Nutr. Metab. Care 19(3): 177-181 (2016), which are hereby incorporated by reference in their entirety). Expression of acetylcholine receptor (AChR) genes is promoted by the PGCla protein (Wheeler et al., “RNA- Binding Proteins Direct Myogenic Cell Fate Decisions,” Elife 11 :e75844 (2022), which is hereby incorporated by reference in its entirety), which is encoded by an ARE-mRNA that is increased in its stability and translation mediated by AUF1 (Abbadi et al., “AUF1 Gene Transfer Increases Exercise Performance and Improves Skeletal Muscle Deficit in Adult Mice,” Mol. Ther. MethodsClin. Dev. 22:222-236 (2021), which is hereby incorporated by reference in its entirety). Moreover, the AUBP HuR was shown to increase the stability and translation of the P-subunit of AChR, which is rate limiting for AChR assembly (Karmouch et al., “AChR P-Subunit mRNAs are Stabilized by HuR in a Mouse Model of Congenital Myasthenic Syndrome with Acetylcholinesterase Deficiency,” Front. Mol. Neurosci. 13:568171 (2020) and Joassard et al., “HuR Mediates Changes in the Stability of AChR P-Subunit mRNAs after Skeletal Muscle Denervation,” J. Neurosci. 35(31): 10949-10962 (2015), which are hereby incorporated by reference in their entirety). AChR genes Chrna, P, 5, and the muscle specific kinase receptor MuSK, which activates muscle contraction, are encoded by mRNAs that have limited AU-rich elements. Also, the gene encoding Rapsyn, which concentrates AChRs in the NMJ to promote muscle contraction, is also stimulated by PGCla, which also stimulates transcription of AChR mRNAs, except perhaps P (Washington et al., “Effects of PGC-la Overexpression on the Myogenic Response During Skeletal Muscle Regeneration,” Sports Med. Health Sci. 4(3): 198- 208 (2022); Arany, “PGC-1 Coactivators and Skeletal Muscle Adaptations in Health and Disease,” Curr. Opin. Genet. Dev. 18(5):426-434 (2008); Kang et al., “Role of PGC-la Signaling in Skeletal Muscle Health and Disease,” Ann. NY. Acad. Sci. 1271(1): 110-117 (2012), which are hereby incorporated by reference in their entirety).

[0193] Therefore, NMJ and associated mRNA and protein levels in the injured TA muscle without and with AAV8 AUF1 prophylaxis were analyzed. AUF1 supplemented TA muscle demonstrated increased levels of all AChR mRNAs except AChR P mRNA at day 7 and day 14, compared to AAV8 controls (FIG. 1 A). There was up to a 60-fold increased level of Rapsyn mRNA by AAV8 AUF1 at day 14 (Fig. IB). MuSK mRNA was increased ~3-fold by AUF1 supplementation (FIG. 1C), as was GABPA, a PGCla responsive transcription factor that promotes mitochondrial biogenesis and Chm gene expression (Mootha et al., “Erralpha and Gabpa / b Specify PGC-1 Alpha-Dependent Oxidative Phosphorylation Gene Expression that is Altered in Diabetic Muscle,” Proc. Natl. Acad. Sci. USA 101 (17): 6570-6575 (2004), which is hereby incorporated by reference in its entirety) (FIG. ID). AUF1 therefore stimulates NMJ mRNA expression both directly and indirectly. Immunofluorescence analysis of synaptophysin protein, which identifies the presynaptic sinus, and AChR in regenerating wounded muscle, were strongly increased compared to control muscle with AUF1 supplementation (FIG. 3), which resembled the uninjured control at day 14. The presence of denervated myofibers by neural cell adhesion molecule (NCAM) staining was quantified (Soendenbroe et al., “Muscle-Nerve Communication and the Molecular Assessment of Human Skeletal Muscle Denervation with Aging,” Am. J. Physiol. Cell. Physiol. 321(2):C317-C329 (2021), which is hereby incorporatedby reference in its entirety). Prophylactic AUF1 supplementation reduced NCAM myofiber staining denervation by 4.5-fold in TA muscle at day 14 compared to AAV control following injury (FIGS. 2A-2B).

[0194] AAV8 AUF1 supplementation at 24 hours post-TA muscle injury also increased expression of AChR, Rapsyn and Musk mRNAs compared to untreated controls, similar to prophylactic administration (FIG. 4A and FIG. 4B). Near normal myofiber architecture, larger fibers and a 4-fold reduction in denervation determined by NCAM staining, was also apparent in TA muscle administered AAV8 AUF1 24 hours post-injury (FIG. 5). AUF1 gene therapy supplementation administered prophylactically or 24 hours post-TA muscle injury therefore increases the expression of NMJ proteins leading to increased muscle re-innervation following injury. Immunofluorescence analysis of synaptophysin protein, which identifies the presynaptic sinus, and AChR in regenerating wounded muscle overlay each other, indicating NMJ reinnervation by AUF1 supplementation therapy (FIG. 6). Muscle function was evaluated by rear leg grip strength testing after injury. Muscle strength tests were performed before injury and every 14 days post injury for 120 days to measure muscle strength. There was a significant (25%) loss of muscle strength following injury in control untreated mice. In contrast, the AUF1 supplementation treatment group demonstrated maintenance of muscle strength with little loss (FIG. 7). In summary, intramuscular administration of AUF1 supplementation, even delayed in administration by 24 hours following severe muscle injury, promotes much more rapid and stronger muscle repair and regeneration with reduced muscle atrophy and necrosis, and preserves muscle from strength loss, indicative of muscle-nerve re-innervation.

[0195] The ability of increased expression of AUF1 to promote greater AChR protein expression was evaluated in AUF1 C2C12 myoblasts induced to differentiate into multinucleated myotubes in culture under low serum. Myoblasts were transduced with LV or LV p45 AUF1 vectors, then induced to differentiate. AUF1 transduction and increased the expression of AUF1 resulted in increased expression of AChR Chrn and Rapsyn mRNAs during myotube differentiation at 96 hours (FIG. 8).

[0196] Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions, and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the claims which follow.

Claims

WHAT IS CLAIMED IS:

1. A method of promoting regeneration of neuromuscular junctions in a subject in need thereof, said method comprising: administering to the subject in need thereof a composition comprising a nucleic acid molecule encoding an AU-rich mRNA binding factor 1 (AUF1) protein or a functional fragment thereof under conditions effective to promote regeneration of neuromuscular junctions in the subject.

2. The method of claim 1, wherein the subject has:(i) degeneration of neuromuscular junctions;(ii) a genetic neuromuscular wasting disorder, optionally wherein the genetic neuromuscular wasting disorder is amyotrophic lateral sclerosis (ALS), Lambert-Eaton Myasthenic Syndrome (LEMS), or a muscular dystrophy, optionally wherein the muscular dystrophy is spinal muscular atrophy (SMA);(iii) a myopathy with a neurologic or inflammatory disorder, optionally wherein the myopathy with a neurologic or inflammatory disorder is inclusion body myositis, Myasthenia gravis, neurogenic amyloidosis, Sjogren's syndrome, multiple sclerosis (MS), ataxia, or rheumatoid arthritis;(iv) an autoimmune disorder, optionally wherein the autoimmune disorder is myasthenia gravis (MG) or Lambert Eaton syndrome;(v) a nerve trauma injury, a nerve cut, or a compression-decompression injury;(vi) a loss of neuromuscular junction as a result of a neurogenic myopathy, optionally wherein the neurologic myopathy is a vacuolar myopathy such as bulbospinal muscular atrophy; and / or(vii) a loss of neuromuscular junctions as a result of a muscular dystrophy, optionally wherein the muscular dystrophy is spinal muscular atrophy (SMA).

3. The method of claim 1 or claim 2, wherein the subject produces autoantibodies against neuromuscular junctions (NMJs), optionally wherein the subject produces autoantibodies against muscle acetylcholine receptor (AChR), muscle specific kinase (MuSK), low density lipoprotein receptor-related protein 4 (LRP4), and / or voltage gated calcium channels (VGCCs).

4. The method of claim 1, wherein a subject has sustained loss of neuromuscular function as a result of chemical or drug toxicity, optionally wherein the chemical or drug is:(i) a pre-synaptic toxin such as botulism toxin or tetanus toxin or a post-synaptic snake venom;(ii) a neurotoxin such as a-bungarotoxin, a-cobratoxin, notexin, or taipoxin;(iii) certain antibiotics such as fluoroquinolone or natural quinoline derivatives such as quinine or quinidine;(iv) aminoglycoside antibiotics; and / or(v) cancer drugs such as vincristine.

5. The method of any one of the preceding claims, wherein said administering is effective to:(i) regenerate neuromuscular junctions in the subject;(ii) increase occurrence of neuromuscular junctions in the subject;(iii) increase occurrence of acetylcholine receptors following muscle injury in the subject;(iv) decrease occurrence of denervated myofibers following muscle injury in the subject;(v) increase occurrence of presynaptic and postsynaptic components following muscle injury in the subject;(vi) decrease acetylcholine degradation in the subject;(vii) increase mitochondrial function in motor neurons in the subject; and / or(viii) increase occurrence of motor neuron synaptic terminals at the neuromuscular junctions in the subject.

6. The method of any one of the preceding claims, wherein said nucleic acid molecule is a vector, optionally wherein the vector is a viral vector, such as wherein the viral vector is an adeno-associated viral (AAV) vector or a lentiviral vector.

7. The method of claim 6, wherein the viral vector is:(i) an adeno-associated viral (AAV) vector, an adeno-associated virus type 8 vector, or an adeno-associated virus type 9 vector or(ii) a lentiviral vector.

8. The method of any one of the preceding claims, wherein said nucleic acid molecule comprises a muscle cell-specific promoter, optionally wherein:(i) the muscle cell-specific promoter is a muscle creatine kinase (MCK) promoter, a smooth muscle 22 (SM22) promoter, a myo-3 promoter, a creatine kinase (CK) 8 promoter, a U6 promoter, a Hl promoter, a Desmin promoter, a Pitx3 promoter, or a skeletal alpha-actin promoter; and / or(ii) the muscle cell-specific promoter is a C5-12 promoter, a CK6-CK9 promoter, a dMCK promoter, a tMCK promoter, a Spc512 promoter, a creatine kinase (CK) 8e promoter, a MHCK7 promoter, or a Sp-3O1 promoter, optionally wherein the muscle cell-specific promoter is a muscle creatine-kinase (tMCK) promoter.

9. The method of any one of the preceding claims, wherein the nucleic acid molecule encodes one or more of p37AUF1, p40AUF1, p42AUF1, or p45AUF1.

10. The method of any one of the preceding claims, wherein the composition comprises a lipid nanoparticle, optionally wherein the lipid nanoparticle is a muscle-tropic lipid nanoparticle, such as wherein the lipid nanoparticle is a nerve-tropic lipid nanoparticle.

11. The method of any one of the preceding claims further comprising: administering a drug or gene therapy effective to treat a neuromuscular disease or disorder in the subject, optionally wherein:(i) the drug or gene therapy comprises an adeno-associated virus 9 vector-based gene therapy;(ii) the drug or gene therapy comprises an antisense oligonucleotide, an siRNA, an shRNA, or miRNA; and / or(iii) the subject has spinal muscular atrophy (SMA) or amyotrophic lateral sclerosis (ALS).

12. The method of claim 11, wherein:(a) the subject has spinal muscular atrophy (SMA) and the gene therapy encodes an SMN1 gene, optionally wherein the gene therapy is ZOLGENSMA® (onasemnogene abeparvovec-xioi), nusinersen (Spinraza), or risdiplam (Evrysdi); or(b) the subject has amyotrophic lateral sclerosis (ALS) associated with a mutation in the superoxide dismutase 1 (SOD1) gene, optionally wherein: the drug comprises an antisense oligonucleotide targeting SOD1 mRNA, such as wherein the drug is tofersen (Qalsody); orthe drug is a benzothiazole, such as wherein the drug is RILUTEX® (riluzole), EXSERVAN™ (riluzole), or TIGLUTIK® (riluzole).

13. The method of claim 11 or claim 12, wherein the composition and the drug or gene therapy are administered simultaneously.

14. The method of claim 11 or claim 12, wherein the composition and the drug or gene therapy are administered sequentially.

15. The method of any one of the preceding claims, wherein said administering the composition and / or administering the drug or gene therapy are carried out by systemic or local administration, optionally wherein said administering is carried out by local administration and wherein said local administration is carried out by intramuscular injection.