Methods for AAV vector re-administration

Abstract

The invention relates to a combination of a recombinant AAVpo1 vector and a recombinant AAV9, AAVrh10 or AAVrh74 vector for use in the gene therapy of muscle and / or nervous system disorders in an individual in need thereof.

Description

METHODS FOR AAV VECTOR RE-ADMINISTRATION FIELD OF THE INVENTION

[0001] The invention pertains to the field of gene therapy. The invention relates to methods for adeno-associated virus (AAV) vector re-administration using a recombinant porcine AAV serotype 1 (AAVpo1) vector in combination with a recombinant AAV serotype 9, rh10, or rh74 (AAV9, AAVrh10, AAVrh74,) vector and their application in gene therapy of diseases, in particular muscle and / or nervous system disorders, more particularly neuromuscular diseases such as genetic neuromuscular diseases. BACKGROUND OF THE INVENTION

[0002] Recombinant Adeno-Associated Virus (rAAV or AAV) vectors are widely used for in vivo gene transfer and clinical trials using AAV vectors are currently taking place for the treatment of a number of diseases.

[0003] AAV is a non-pathogenic virus belonging to the genus Dependoparvovirus within the family Parvoviridae. AAV is a non-enveloped virus composed of a capsid of ~25 nm of diameter and a single strand DNA of 4.7 kb. The genome carries two genes, rep and cap, flanked by two palindromic regions named Inverted terminal Repeats (ITR) that serve as the viral origins of replication and the packaging signal. The cap gene codes for three structural proteins VP1, VP2 and VP3 that compose the AAV capsid. VP1, VP2 and VP3 share the same C-terminal end which is all of VP3. Using AAV2 has a reference, VP1 has a 735 amino acid sequence (GenBank YP_680426); VP2 (598 amino acids) starts at the Threonine 138 (T138) and VP3 (533 amino acids) starts at the methionine 203 (M203). The rep gene encodes four proteins required for viral replication Rep78, Rep68, Rep52 and Rep40. Recombinant AAV vectors encapsidate an ITR-flanked rAAV genome in which a therapeutic gene expression cassette replaces the AAV protein coding-sequences.

[0004] Tissue specificity is determined by the capsid serotype and commonly used AAV serotypes isolated from human (AAV2, 3, 5, 6) and non-human primates (AAV1, 4, 7-11) can transduce specific organs more efficiently than others, such as AAV6, AAV8, AAV9 and AAV-rh74 in muscle tissue and AAV2, AAV9, AAVrh10, AAVcy.10, AAV-PHP.B, AAV-PHP.EB and clade F AAVHSC such as AAVHSC7, AAVHSC15 and AAVHSC17 in nervous tissue.

[0005] AAV vector immunogenicity represents a major limitation to gene transfer with AAV vectors. On one end, there is a pre-existing immunity to commonly used AAV vector serotypes isolated from human and non-human primates; for example AAV2 is seroprevalent in up to 80 % of the human population (Fu et al., Hum Gene Ther Clin Dev., 2017 Dec;28(4):187-196; Stanford et al., Res Pract Thromb Haemost., 2019, 3: 261-267). On the other end, the high-level neutralising antibodies generated after the administration of commonly used AAV vector serotypes, prevents the possibility of vector re-administration.

[0006] Several attempts were made to re-administer an AAV vector in the presence of anti-AAV antibodies such as plasmapheresis or treatment with IgG-cleaving endopeptidase (IdeS) before second dose of vector injection. However, this protocol is complicated.

[0007] Recombinant AAV vectors have been generated using capsids from different porcine AAVs (AAVpo1, po2.1, po4 to 6) and following systemic administration in mice, strong transgene expression in all major skeletal muscle types combined with poor transduction of other tissues including complete detargeting from the liver was reported for AAVpo1 (Bello et al., Gene Therapy, 2009, 16, 1320-1328. doi: 10.1038 / gt.2009.82; Bello et al., Sci Rep., 2014, 4, 6644, doi: 10.1038 / srep06644; Tulalamba et al., Gene Therapy, 2019, doi.org / 10.1038 / s41434-019-0106-3; WO 2009 / 030025). A peptide-modified AAVpo1 (AAVpo1.A1) is further capable of transducing the central nervous system (brain and spinal cord) and achieving transgene expression levels in different muscle groups and in the central nervous system that were at least equivalent if not superior to that of AAV9 vector while at the same time being detargeted from the liver (WO 2021 / 219762).

[0008] There is a need in the art for methods for adeno-associated virus (AAV) vector re- administration. SUMMARY OF THE INVENTION

[0009] The inventors have shown that contrary to what was disclosed previously (Bello et al., Tulalamba et al., precited; WO 2009 / 030025), there is a cross-neutralization between the porcine AAV serotype and some of the current AAV vector serotypes isolated from humanand non-human primates. In addition, AAVpo1 is neutralized by human immunoglobulins but the neutralization titers of the porcine AAV serotype are low compared to that of the current AAV serotypes. Despite of these findings, the inventors have shown that AAV vector re- administration should be possible after an initial administration of AAV vector using appropriate combinations of AAVpo1 and current AAV vector serotypes.

[0010] The examples of the present application show that AAVpo1 is not neutralized by antibodies produced after AAV8, AAV9, AAVrh74 or AAVrh10 injection (Figure 1 and Figure 3), allowing a re-administration with AAVpo1 vector after an initial injection of AAV8, AAV9, AAVrh74 or AAVrh10 vector (Figure 6 and Figure 7). In contrast, re- administration with AAV8, AAV9 or AAVrh10 vector after an initial injection of AAVpo1 vector, resulted in undetectable levels of skeletal muscle transduction due to the presence of neutralizing antibodies, even at low levels for AAV8 or AAVrh10, prior to the second injection (Figure 2, Figure 4, Figure 6 and Figure 7).

[0011] The invention relates to a combination of a recombinant porcine adeno-associated virus serotype 1 (AAVpo1) vector and a recombinant adeno-associated virus serotype 9, rh10 or rh74 (AAV9, AAVrh10, AAVrh74) vector, for use in the gene therapy of diseases, preferably wherein the AAV9, AAVrh10 or AAVrh74 vector is administered at an initial time point and the AAVpo1 vector is administered at a later time point.

[0012] In some embodiments, the AAVpo1, AAV9, AAVrh10 or AAVrh74 serotype is selected from the group consisting of: AAV capsids comprising a sequence having at least 95 % identity with anyone of SEQ ID NO: 1 and 3 to 5 and hybrid or peptide-modified derivatives thereof having at least 90 % identity with any one of SEQ ID NO: 1 and 3 to 5.

[0013] In some preferred embodiments, the peptide-modified AAV serotype comprises a peptide comprising at least the sequence RGD, preferably. a 7mer peptide comprising at least the sequence RGD. In some preferred embodiments, the peptide-modified AAV serotype comprises a peptide selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82; preferably SEQ ID NO: 6, 15, 33 to 60 and 77 to 82; more preferably SEQ ID NO: 6, 15, 36, 44, 50, 56, 58, 60, 77 and 79.

[0014] In some preferred embodiments, the AAVpo1 serotype has at least 95 % identity with SEQ ID NO: 61 and comprises the peptide A1 (SEQ ID NO: 15). In some preferredembodiments, the AAV9 serotype has at least 95 % identity with SEQ ID NO:3 and comprises a peptide selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably selected from the group consisting of SEQ ID NO: 6, 15, 33 to 60 and 77 to 82 and more preferably SEQ ID NO: 6, 15, 36, 44, 50, 56, 58, 60, 77 and 79. In some preferred embodiments, the AAV9 serotype is hybrid AAV9rh74 having at least 95 % identity with SEQ ID NO: 62; preferably comprising the peptide P1 (SEQ ID NO: 6) or a peptide of any one of SEQ ID NO: 77 to 82; more preferably having at least 95 % identity with SEQ ID NO: 64 or 65 and comprising the peptide P1 (SEQ ID NO: 6) or having at least 95 % identity with SEQ ID NO: 83 and comprising the peptide SEQ ID NO: 77.

[0015] In some embodiments, the recombinant AAVpo1 vector and the recombinant AAV9, AAVrh74 or AAVrh10 vector encode a transgene of interest for therapy. In some particular embodiments, the transgene encoded by the recombinant AAVpo1 vector and the transgene encoded by the recombinant AAV9, AAVrh10 or AAVrh74 vector are the same. In some other particular embodiments, the transgene encoded by the recombinant AAVpo1 vector and the transgene encoded by the recombinant AAV9, AAVrh10 or AAVrh74 vector are different.

[0016] In some embodiments, the transgene of interest for therapy is selected from the group consisting of: (i) therapeutic genes; (ii) genes encoding therapeutic proteins or peptides such as therapeutic antibodies or antibody fragments and genome editing enzymes; and (iii) genes encoding therapeutic RNAs such as interfering RNAs, guide RNAs for genome editing and antisense RNAs capable of exon skipping.

[0017] In some particular embodiments, the recombinant AAV serotype for the initial administration and the recombinant AAV serotype for the later administration are chosen respectively from : (i) natural AAV9 capsid of SEQ ID NO: 3 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61 ; (ii) AAVrh10 capsid, in particular natural AAVrh10 capsid of SEQ ID NO: 4 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61 (iii) peptide-modified AAV9 capsid comprising a peptide chosen from SEQ ID NO: 33 to 60 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61; (iv) AAVrh74 capsid, inparticular natural AAVrh74 capsid of SEQ ID NO: 5 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61.

[0018] In some embodiments, the combination is administered by systemic route, preferably intravascular route; more preferably intravenous route.

[0019] In some embodiments, the combination according to the present disclosure is used in the gene therapy of muscle and / or nervous system diseases or disorders.

[0020] In some embodiments, the target cells for gene therapy are in the nervous system (central and / or peripheral), muscles (cardiac, smooth and / or skeletal), or a combination thereof.

[0021] In some embodiments, the disease is selected from the group consisting of: neurological diseases, muscular diseases, and combination thereof. Preferably, the disease is a neuromuscular disease affecting the nervous system, preferably a genetic neuromuscular disease affecting the nervous system. In some preferred embodiments, the genetic neuromuscular disease is selected from the group comprising : (i) myopathies, such as congenital myopathies, myasthenic syndromes, metabolic myopathies, distal myopathies, muscular dystrophies with or without cardiomyopathy; and (ii) spinal muscular atrophies (SMAs) and motor neuron diseases; preferably SMN1-related spinal muscular atrophy, spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME), spinobulbar muscular atrophy (SBMA) and amyotrophic lateral sclerosis (ALS).

[0022] In some preferred embodiments, the target gene for gene therapy is selected from the group comprising: DMD, DYSF, FKRP, α-sarcoglycan (SGCA), β-sarcoglycan (SGCB), γ-sarcoglycan (SGCG), Calpain 3 (CAPN3), Anoctamin 5 (ANO5), MTM1, DNM2, BIN1, GAA, AGL, ColQ, DOK7, SMN1, ASAH1, MCEP2, and AR genes.

[0023] The invention also relates to a product containing a recombinant AAVpo1 vector and a recombinant AAV9, AAVrh10 or AAVrh74 vector as a combined preparation for sequential use in the treatment of a disorder according to the present disclosure.DETAILED DESCRIPTION OF THE INVENTION

[0024] The invention relates to a combination of a recombinant porcine adeno-associated virus serotype 1 (AAVpo1) vector and a recombinant adeno-associated virus serotype 9, rh10 or rh74 (AAV9, AAVrh10, AAVrh74) vector for use in the gene therapy of diseases in an individual in need thereof, preferably wherein the AAV9, AAVrh10, or AAVrh74 vector is administered at an initial time point and the AAVpo1 vector is administered at a later time point.

[0025] As used herein, the term “AAV vector” refers to an AAV vector particle.

[0026] As used herein, the term “recombinant porcine AAV serotype 1 (AAVpo1) vector” or AAVpo1 vector refers to an AAV vector comprising a porcine AAV serotype 1 capsid protein. The term “recombinant AAV serotype 9, rh10, rh74 (AAV9, AVrh10, AAVrh74) vector” or AAV9, AAVrh10 AAVrh74 vector refers to an AAV vector comprising an AAV serotype 9, rh10, rh74 capsid protein.

[0027] As used herein, the term “AAV serotype” refers to an AAV capsid serotype. AAV serotype 9, rh10, rh74 refers to AAV9, AAVrh10 or AAVrh74 capsid. AAVpo1 serotype refers to AAVpo1 capsid. AAV serotype includes natural AAV (i.e. wild-type AAV) and artificial AAV serotypes such as variants and hybrid capsids derived from natural AAV serotypes, as well as peptide-modified AAV serotypes derived from natural, variant or hybrid AAV serotypes. AAV serotype refers to a functional AAV capsid which is able to transduce cells of the target tissue or organ and express a transgene in said target tissue or organ.

[0028] As used herein, AAV capsid refers to AAV, VP1, VP2 and / or VP3 protein.

[0029] As used herein, the term “target tissue or organ” refers to an individual tissue or organ or plurality of tissues or organs that can be targeted by the combination of recombinant AAVpo1 serotype vector and recombinant AAV9, AAVrh10 or AAVrh74 serotype vector according to the present disclosure. The target tissue or organ is targeted for gene therapy, i.e., for treating diseases by sequential administration of the combination of AAV vectors according to the present disclosure.

[0030] As used herein, “for use in the gene therapy of diseases or disorders, means “for use in the treatment of diseases or disorders by gene therapy” or “for use in the treatment by gene therapy of diseases or disorders.

[0031] As used herein, ‘disease” or “disorder” refer to a disease that can be treated by gene therapy using the combination of AAV vectors according to the present disclosure. A disease includes in particular, a disease associated with a gene mutation and therefore eligible to AAV gene therapy.

[0032] “a”, “an”, and “the” include plural referents, unless the context clearly indicates otherwise. As such, the term “a” (or “an”), “one or more” or “at least one” can be used interchangeably herein; unless specified otherwise, “or” means “and / or”.

[0033] AAVpo1 (GenBank accession number FJ688147 as accessed on 24 July 2016) comprises the Cap gene from position 780 to 2930 of the partial viral genome sequence (2977 bp): VP1 CDS is from positions 780 to 2930; VP2 CDS is from positions 1188 to 2930 and VP3 CDS is from positions 1329 to 2930. The AAVpo1 capsid protein (VP1) has the sequence GenBank accession number ACN42940.1 as accessed on 24 July 2016 or SEQ ID NO: 1. AAV9 capsid corresponds to the amino acid sequence GenBank accession number AY530579.1 or protein ID number AAS99264.1 accessed on 24 June 2004 (SEQ ID NO: 3). AAVrh10 capsid (GenBank accession number AY243015.1 or protein ID number AAO88201.1 accessed on 14 May 2003) corresponds to the amino acid sequence SEQ ID NO: 4. AAVrh74 capsid corresponds to the amino acid sequence SEQ ID NO: 5.

[0034] AAVpo1, AAV9, AAVrh74 or AAVrh10 serotype includes the natural (wild- type) serotype as listed above (SEQ ID NO: 1 and 3 to 5) as well as any artificial serotype including any variant, hybrid and / or peptide-modified AAV capsid derived from said serotype. The invention encompasses the use of AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid or serotype having at least at least 95%, 96%, 97%, 98% or 99% identity with AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5). The invention also encompasses the use of hybrid and / or peptide-modified AAV capsids derived from said AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid or serotype, wherein the hybrid and / or peptide-modified AAV capsid has at least at least 90% identity with AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5).The hybrid and / or peptide-modified AAV capsid may have 91%, 92%, 93%, 94%, 95% or more (96%, 97%, 98% or 99%) identity with AAVpo1, AAV9, AAVrh74, AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5). A peptide-modified AAV capsid according to the invention: (i) is derived from an initial AAV capsid (without the peptide(s)) having at least at least 95%, 96%, 97%, 98% or 99% identity with AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5) and (ii) has at least 90% identity with AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5). A hybrid AAV capsid according to the invention: (i) is derived from at least two different AAV serotypes including at least one AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid or serotype having at least 95%, 96%, 97%, 98% or 99% identity with AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5) and (ii) has at least 90% identity with said at least one AAVpo1, AAV9, AAVrh74 or AAVrh10 capsid sequence as listed above (SEQ ID NO: 1 and 3 to 5). An artificial serotype according to the invention has a seroneutralization profile that is similar to that of the natural serotype it is derived from. This means that the artificial serotype is neutralized by AAV antibodies with titers similar to those of the natural serotype. Preferably, similar titers differ from each other by a dilution factor of less than 2; preferably less than 3. A 3.16-fold (100.5- fold) dilution is called a half-logarithmic dilution or half-log dilution.

[0035] The term “identity” refers to the sequence similarity between two polypeptide molecules or between two nucleic acid molecules. When a position in both compared sequences is occupied by the same base or same amino acid residue, then the respective molecules are identical at that position. The percentage of identity between two sequences corresponds to the number of matching positions shared by the two sequences divided by the number of positions compared and multiplied by 100. Generally, a comparison is made when two sequences are aligned to give maximum identity. The identity may be calculated by alignment using, for example, the GCG (Genetics Computer Group, Program Manual for the GCG Package, Version 7, Madison, Wisconsin) pileup program, or any of sequence comparison algorithms such as BLAST, FASTA or CLUSTALW.

[0036] In some embodiments, the AAVpo1, AAV9, AAVrh10 or AAVrh74 serotype (natural, variant or hybrid AAV capsid) is peptide-modified, in particular with a peptide which enhances transduction of cells of the target tissue or organ. Libraries of AAV capsid variantsdisplaying short random peptides on the surface of various AAV serotypes have been generated to screen for gene therapy vectors with altered cell specificities and / or transduction efficiencies (Review in Büning et al., Molecular Therapy: Methods & Clinical Development, 2019, 12, 248). Various peptides able to target AAV vectors to a desired organ (targeting peptides) obtained by library screening are disclosed in the art (Michelfelder et al., PLoS ONE, 2009, 4, e5122; Kienle EC (Dissertation for the degree of Doctor of natural Sciences, Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany, 2014); WO 2019 / 207132; Börner et al., Molecular Therapy, April 2020, 28, 1017-1032; Weinmann et al., Nature communications, 2020, 11, 5432; Tabebordbar et al., Cell, 2021, 184, 4919-4938). Peptides targeting integrin alpha V beta 6 which improve AAV vector myotropism are disclosed in Ai Vu Hong et al., Nature Communications, 2024, 15, 7965- and WO 2023 / 237748. The targeting peptide consists generally of a sequence of up to 30 amino acids. The peptide may consist of 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 amino acids. In some embodiments, the peptide consists of a sequence of up to 15 amino acids, preferably 3 to 10 amino acids, in particular 7 or 10 amino acids.

[0037] Non-limiting examples of targeting peptides include the peptides disclosed in Börner et al (Table S2 and Table S3) ; Weinmann et al., Tabebordbar et al.; Ai Vu Hong et al. and WO 2023 / 237748: P1 (RGDLGLS), P2 (CDCRGDCFC), P3 (RGDAVGV), P4 (NDVRSAN), P5 (NDVRAVS), P6 (CNHRYMQMC), P7 (SPGARAF), P8 (DGPWRKM), P9 (FGQKASS), A1 (MPLGAAG), A2 (NYSRGVD), A3 (GSVPRLG), A4 (PVVPRPA), A5 (GERAKPA), A6 (NEARVRE), D1 (NSSRDLG), D2 (MVNNFEW), T1 (SEGLKNL), T2 (PSVPRPP), K1 (SLRSPPS), K2 (NFTRLSA), L1 (GDVGPPG), L2 (NDVRPER), L4 (NRVEEKL), L5 (TTSVRPA), L6 (HLHGRPA), Kera2 (PRGDLAP), RGDLRVS, 1C (RGDLSTP), 1B (RGDLNQY), 1A (RGDLTTP), 1F (RGDATEL), 1D (RGDQLYH), RGDVAAK, 1E (RGDTMSK), 1G (RGDMINT), 1H (RGDLNDS), RGDTMNY, 2A (GPGRGDQTTL), 2B (AEGRGDQYTR), 2C (ATGRGDLGQA), 2D (AVARGDQGLI), 2E (NISRGDQGYQ), 2F (APARGDQGSQ), 3A (RGDYVGL), 3B (RGDYSGL), 3C (RGDYSSV), 3D (RGDYREL), 3E (RGDHGVL), 3F (RGDHASW), 4A (SNSRGDYNSL), 4B (STVRGDYTS), 4C (QERRGDYTSM), 4D (ASTRGDHGVL), 4E (ENRRGDFNNT), RGDLXXL / I wherein XX may be any pair of amino acids, which binds with high affinity to the integrin heterodimer αVβ6 (ITGAV-B6), in particular 4um9 (RGDLGRL),4um9_modified (RGDLGEL), 5ffo (RGDLATI), 5ffo_modified (RGDLAEL), 5nem (RGDLQVL) and 5nem_modified (RGDLAEI), corresponding to SEQ ID NO: 6 to 60 and 77 to 82.

[0038] AAV capsids comprising the insertion of a peptide comprising an RGD motif, which is known to bind several different cell-surface integrins, have been reported to improve gene delivery in muscle following systemic administration. Peptides containing RGD motif are disclosed in Tabebordbar et al., Cell, 2021, 184, 4919-4938; Ai Vu Hong et al. and WO 2023 / 237748; and include: RGDLSTP (1C), RGDLNQY (1B), RGDLTTP (1A), RGDATEL (1F), RGDQLYH (1D), RGDVAAK, RGDTMSK (1E), RGDMINT (1G), RGDLNDS, RGDTMNY, 2A, 2B, 2C, 2D, 2E, 2F, 3A, 3B, 3C, 3D, 3E, 3F, 4A, 4B, 4C, 4D, 4E, RGDLGRL (4um9), RGDLGEL (4um9_modified), RGDLATI (5ffo), RGDLAEL (5ffo_modified), RGDLQVL (5nem) and RGDLAEI (5nem_modified). The most efficient AAV capsids for muscle transduction in mice, non-human primates and / or human primary myotubes display a peptide comprising a RGDL motif : RGDLGLS or P1 (AAVMYO or AAV9P1), RGDLTTP (MyoAAV 1A), GPGRGDQTTL (MyoAAV 2A), SNSRGDYNSL (MyoAAV 4A), ENRRGDFNNT (MyoAAV 4E), SAQRGDYVGL (MyoAAV 3A), QERRGDYTSM (MyoAAV 4C) inserted into the variable region VIII (WO 2019 / 207132; Weinmann et al., Nature communications, 2020, 11, 5432; Tabebordbar et al., Cell, 2021, 184, 4919-4938); or RGDLGRL (4um9), RGDLGEL (4um9_modified), RGDLATI (5ffo), RGDLAEL (5ffo_modified), RGDLQVL (5nem) and RGDLAEI (5nem_modified) inserted into the variable region IV (Ai Vu Hong et al. and WO 2023 / 237748) . In some preferred embodiments, the AAVpo1, AAV9, AAVrh10 or AAVrh74 serotype is modified with a peptide selected from the group consisting of : P1 (RGDLGLS), A1 (MPLGAAG), MyoAAV peptide (SEQ ID NO: 33 to 60) and SEQ ID NO: 77 to 82, in particular SEQ ID NO: 36 (MyoAAV 1A peptide), SEQ ID NO: 44 (MyoAAV 2A peptide), SEQ ID NO: 50 (MyoAAV 3A peptide), SEQ ID NO: 56 (MyoAAV 4A peptide), SEQ ID NO: 58 (MyoAAV 4C peptide), SEQ ID NO: 60 (MyoAAV 4E peptide), SEQ ID NO: 77 (4um9 peptide) and SEQ ID NO: 79 (5ffo peptide).

[0039] In some embodiments, the AAVpo1 serotype is natural AAVpo1; variant AAVpo1, hybrid AAVpo1 or peptide-modified derivative thereof, preferably natural AAVpo1 or peptide-modified AAVpo1. In some particular embodiments, the AAVpo1 serotype (natural, hybrid or variant capsid) has at least 95 % identity with SEQ ID NO: 1. Insome particular embodiments, the peptide comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82; preferably SEQ ID NO: 15, 33 to 60 and 77 to 82. In some more particular embodiments, the peptide comprises or consists of the sequence MPLGAAG (peptide A1 or SEQ ID NO: 15). In some particular embodiments, the AAVpo1 serotype has at least 90 % identity with SEQ ID NO: 1, preferably at least 95 % identity with SEQ ID NO: 1, and comprises a peptide chosen from SEQ ID NO: 15, 33 to 60 and 77 to 82, preferably SEQ ID NO: 15. In some preferred embodiments, said peptide- modified AAVpo1 capsid protein comprises a sequence selected from the group consisting of the sequence SEQ ID NO: 61 and the sequences having at least 95%, 96%, 97%, 98% or 99% identity with SEQ ID NO: 61 which comprise said peptide of SEQ ID NO: 15, and the fragment thereof corresponding to VP2 or VP3 capsid protein. SEQ ID NO: 61 corresponds to the AAVpo1 capsid modified by the peptide A1, named AAVpo1.A1, disclosed in WO 2021 / 219762. VP2 corresponds to the amino acid sequence from K136 to the end of SEQ ID NO: 61. VP3 corresponds to the amino acid sequence from M184 to the end of SEQ ID NO: 61. In some preferred embodiments, said peptide-modified AAVpo1 capsid protein comprises the sequence SEQ ID NO: 61, or a fragment thereof corresponding to VP2 or VP3 capsid protein.

[0040] In some embodiments, the AAV9 serotype is natural (wild-type) AAV9 or peptide-modified AAV9 comprising a peptide chosen from SEQ ID NO: 3 to 60 and 77 to 82, preferably the P1 peptide, the A1 peptide, a MyoAAV peptide chosen from SEQ ID NO: 33 to 60, or a RGDLXXL / I peptide chosen from SEQ ID NO: 77 to 82, in particular SEQ ID NO: 36, 44, 50, 56, 58, 60, 77 and 79. P1-modified AAV9 capsid (AAV9P1 or AAVMYO) is disclosed in WO 2019 / 207132 and corresponds to the amino acid sequence SEQ ID NO:63.

[0041] In some embodiments, the AAV9 serotype is hybrid AAV9 or peptide-modified derivative thereof. In particular embodiments, the peptide comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably SEQ ID NO: 6, 15, 33 to 60 and 77 to 82. In particular embodiments, AAV9 serotype is hybrid AAV9.rh74, P1-modified AAV9.rh74 or AAV9.rh74 modified with RGDLXXL / I peptide 4um9, 4um9_modified, 5ffo, 5ffo_modified, 5nem or 5nem_modified. AAV9.rh74 modified with RGDLXXL / I peptide is preferably modified with peptide 4um9. AAV9.rh74 capsid is disclosed in WO 2019 / 193119 and corresponds to the amino acid sequence SEQ ID NO: 62.P1-modified AAV9rh74 capsids are disclosed in WO2022 / 053630 and WO 2020 / 200499 and correspond to SEQ ID NO: 64 and SEQ ID NO: 65, respectively. AAV9.rh74 modified with RGDLXXL / I peptide 4um9, 4um9_modified, 5ffo, 5ffo_modified, 5nem or 5nem_modified are disclosed in WO 2023 / 237748 and correspond to SEQ ID NO: 83 to 88, respectively. SEQ ID NO: 83 is LICA1.

[0042] In some embodiments, the AAV9 serotype is AAV9 variant or peptide-modified derivative thereof. In particular embodiments, the peptide comprises or consists of a sequence selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably SEQ ID NO: 6, 15, 33 to 60, and 77 to 82. AAV9 variant includes in particular: AAV-PHP such as AAV-PHP.A, AAV-PHP.B, AAV-PHP.eB, AAV-PHP.S; AAV9BI; AAV9LD; AAVS1and AAVS10 (El Andari et al., Science Advances, 2022, 8, 1-21; WO2019207132A1). Peptide- modified AAV9 variant includes in particular AAVMYO2 (SEQ ID NO: 66) and AAVMYO3 (SEQ ID NO: 67), corresponding to P1-modified AAVS1 and AAVS10 capsids respectively (El Andari et al.; WO2019207132A1).

[0043] In some particular embodiments, the AAV9 serotype has at least 90 % identity with SEQ ID NO: 3; preferably at least 95 % identity with SEQ ID NO: 3. In some particular embodiments, the AAV9 serotype has at least 90 % identity with SEQ ID NO: 3, preferably at least 95 % identity with SEQ ID NO: 3, and comprises a peptide comprising or consisting of a sequence selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably SEQ ID NO: 6, 15, 33 to 60 and 77 to 82; more preferably the peptide P1 (SEQ ID NO: 6); more preferably comprising a sequence having at least 95 % identity with any one of SEQ ID NO: 63, 66 or 67 which comprises said P1 peptide (SEQ ID NO: 6); still more preferably comprising any one of SEQ ID NO: 63, 66 and 67.

[0044] In some particular embodiments, AAV9 serotype is hybrid AAV9rh74, preferably having at least 95 % identity with SEQ ID NO: 62 and / or comprising a peptide comprising or consisting of a sequence selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably SEQ ID NO: 6, 15, 33 to 60 and 77 to 82; more preferably the peptide P1 (SEQ ID NO: 6) or a RGDLXXL / I peptide chosen from 4um9, 4um9_modified, 5ffo, 5ffo_modified, 5nem and 5nem_modified, preferably peptide 4um9 (SEQ ID NO: 77). In some preferred embodiment hybrid AAV9rh74 serotype comprises a sequence having at least 95 % identity with SEQ ID NO: 62, or a sequence having at least 95 % identity with SEQ IDNO: 64 or 65 which comprises said peptide P1; more preferably comprising any one of SEQ ID NO: 62, 64 or 65. In some other preferred embodiment hybrid AAV9rh74 serotype comprises a sequence having at least 95 % identity with SEQ ID NO: 62, or a sequence having at least 95 % identity with SEQ ID NO: 83 to 88 which comprises said RGDLXXL / I peptide chosen from 4um9, 4um9_modified, 5ffo, 5ffo_modified, 5nem and 5nem_modified, respectively; more preferably comprising any one of SEQ ID NO: 83 to 88; still more preferably SEQ ID NO: 83.

[0045] In some embodiments, the AAVrh10 serotype is natural AAVrh10; variant AAVrh10, hybrid AAVrh10 or peptide-modified derivative thereof. In particular embodiments, AAVrh10 serotype (natural, hybrid or variant capsid) comprises a peptide chosen from SEQ ID NO: 6 to 60 and 77 to 82, preferably the P1 peptide, the A1 peptide or a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 and the RGDLXXL / I peptide chosen from SEQ ID NO: 77 to 82, in particular SEQ ID NO: 36, 44, 50, 56, 58, 60, 77 and 79. In some particular embodiments, the AAVrh10 serotype has at least 95 % identity with SEQ ID NO: 4. In some particular embodiments, the AAVrh10 serotype has at least 90 % identity with SEQ ID NO: 4, preferably at least 95 % identity with SEQ ID NO: 4, and comprises the peptide P1 (SEQ ID NO: 6), a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 or a RGDLXXL / I peptide chosen from SEQ ID NO: 77 to 82, in particular SEQ ID NO: 36, 44, 50, 56, 58, 60, 77 and 79.

[0046] In some embodiments, the AAVrh74 serotype is natural AAVrh74; variant AAVrh74, hybrid AAVrh74 or peptide-modified derivative thereof. In particular embodiments, AAVrh74 serotype (natural, hybrid or variant capsid) comprises a peptide chosen from SEQ ID NO: 6 to 60 and 77 to 82, preferably the P1 peptide, the A1 peptide, a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 or a RGDLXXL / I peptide chosen from SEQ ID NO: 77 to 82, in particular SEQ ID NO: 36, 44, 50, 56, 58, 60, 77 and 79. In particular embodiments, AAVrh74 serotype is hybrid AAV9.rh74 or peptide-modified derivative thereof as disclosed above. In some particular embodiments, the AAVrh74 serotype has at least 95 % identity with SEQ ID NO: 5. In some particular embodiments, the AAVrh74 serotype has at least 90 % identity with SEQ ID NO: 5, preferably at least 95 % identity with SEQ ID NO: 5, and comprises the peptide P1 (SEQ ID NO: 6), a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 or a RGDLXXL / I peptide chosen from SEQ ID NO: 77 to 82, inparticular SEQ ID NO: 36, 44, 50, 56, 58, 60, 77 and 79; more preferably comprising SEQ ID NO: 6 or SEQ ID NO: 77. In some particular embodiments, the AAVrh74 serotype is hybrid AAV9rh74, preferably having at least 95 % identity with SEQ ID NO: 62 or having at least 95 % identity with SEQ ID NO: 64 or 65 and comprising peptide P1 (SEQ ID NO: 6); more preferably comprising a sequence selected from SEQ ID NO: 62, 64 and 65.In some other preferred embodiment hybrid AAV9rh74 serotype comprises a sequence having at least 95 % identity with SEQ ID NO: 62, or a sequence having at least 95 % identity with SEQ ID NO: 83 to 88 which comprises said RGDLXXL / I peptide chosen from 4um9, 4um9_modified, 5ffo, 5ffo_modified, 5nem and 5nem_modified, respectively; more preferably comprising any one of SEQ ID NO: 83 to 88; still more preferably SEQ ID NO: 83.

[0047] In some embodiments, the combination of rAAV for the initial and the later administrations comprises respectively: (i) natural AAV9 capsid and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (ii) AAVrh10 capsid, in particular natural AAVrh10 capsid and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (iii) peptide-modified AAV9 capsid comprising a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (iv) AAVrh74 capsid, in particular natural AAVrh74 capsid and peptide- modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (v) peptide-modified AAV9 capsid comprising the peptide P1 (AAV9P1 or AAVMYO) and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (vi) peptide-modified AAV9 variant capsid comprising the peptide P1 named AAVS1P1 (AAVMYO2) and peptide- modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (vii) peptide-modified AAV9 variant capsid comprising the peptide P1 named AAVS10P1 (AAVMYO3) and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (viii) hybrid AAV9rh74 capsid including peptide modified hybrid AAV9rh74 capsid, in particular comprising the peptide P1 (AAV9rh74P1) or the peptide 4um9 (LICA1) and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1). Preferably, the combination of rAAV for the initial and the later administrations comprises respectively: (i) natural AAV9 capsid and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (ii) AAVrh10 capsid, in particular natural AAVrh10 capsid and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1); (iii) peptide-modified AAV9 capsid comprising a MyoAAV peptide chosen from SEQ ID NO: 33 to 60 and peptide-modified AAVpo1 capsidcomprising the peptide A1 (AAVpo1A1); (iv) AAVrh74 capsid, in particular natural AAVrh74 capsid and peptide-modified AAVpo1 capsid comprising the peptide A1 (AAVpo1A1).

[0048] In some particular embodiments, the invention also relates to a recombinant porcine adeno-associated virus serotype 1 (AAVpo1) vector for use in the gene therapy of diseases according to the present disclosure, wherein the AAVpo1 vector is administered to a patient having a natural immunity to an AAV9, AAVrh10 and / or AAVrh74 vector according to the present disclosure. As used herein, a patient having a natural immunity to an AAV9, AAVrh10 and / or AAVrh74 vector relates to a patient having pre-existing neutralizing antibodies against AAV9, AAVrh10 or AAVrh74 resulting from natural infection with adeno- associated virus (AAV). The presence and / or level of neutralizing antibodies against AAV9, AAVrh10 or AAVrh74 in patient plasma or serum may be determined using appropriate neutralization assays that are well-known in the art and disclosed in the present application. One example of neutralization assay is disclosed in Meliani et al., Hum. Gene Ther. Methods, 2015, 26, 45-53).

[0049] The genome of the AAV vectors for use in combination according to the present disclosure either be a single-stranded or self-complementary double-stranded genome (McCarty et al, Gene Therapy, 2003, Dec.,10(26), 2112-2118). Self-complementary vectors are generated by deleting the terminal resolution site (trs) from one of the AAV terminal repeats. These modified vectors, whose replicating genome is half the length of the wild-type AAV genome have the tendency to package DNA dimers. The AAV genome is flanked by ITRs. In particular embodiments, the AAV vector is a pseudotyped vector, i.e. its genome and capsid are derived from AAVs of different serotypes. In some embodiments, the vector comprises AAV2 rep proteins and / or AAV2 ITRs.

[0050] The AAV vectors for use in combination according to the present disclosure are produced by standard methods for producing AAV vectors that are well-known in the art (Review in Aponte-Ubillus et al., Applied Microbiology and Biotechnology, 2018, 102: 1045- 1054). Briefly, following co-transfection with expression plasmid(s) for AAV Rep and Capsid proteins and plasmid containing recombinant AAV vector genome comprising the gene of interest inserted in an expression cassette, flanked by AAV ITRs, in the presence of sufficient helper function to permit packaging of the rAAV vector genome into AAV capsid particle,the cells are incubated for a time sufficient to allow the production of AAV vector particles, the cells are then harvested, lysed, and AAV vector particles are purified by standard purification methods such as affinity chromatography or Iodixanol or Cesium Chloride density gradient ultracentrifugation.

[0051] The AAV vector particle usually packages a gene (i.e. a transgene) of interest for therapy. By “gene of interest for therapy”, “gene of therapeutic interest”, “gene of interest” or “heterologous gene of interest”, it is meant a therapeutic gene or a gene encoding a therapeutic protein, peptide or RNA or a combination thereof.

[0052] In some embodiments, the transgene encoded by the recombinant AAVpo1 vector and the transgene encoded by the recombinant AAV9, AAVrh10 or AAVrh74 vector are the same. In some other embodiments, the transgene encoded by the recombinant AAVpo1 vector and the transgene encoded by the recombinant AAV9, AAVrh10, or AAVrh74 vector are different. The invention encompasses multiple vector systems such as dual vector systems, wherein the combination of vectors encodes overlapping fragments of the transgene that form a complete transgene after co-delivery of the combination of vectors into cells.

[0053] The gene of interest is any nucleic acid sequence capable of modifying a target gene or target cellular pathway, in cells of target tissue or organ. For example, the gene may modify the expression, sequence or regulation of the target gene or cellular pathway.

[0054] In some particular embodiments, the target tissue or organ comprises muscles and / or nervous system.

[0055] As used herein, the term “muscle” refers to cardiac muscle (i.e. heart), smooth muscle and skeletal muscle. The term “muscle cells” refers to myocytes, myotubes, myoblasts, and / or satellite cells.

[0056] As used herein, the term “nervous system “, refers to both the central (CNS) and peripheral (PNS) nervous system. The term “central nervous system or CNS” refers to the brain, spinal cord, retina, cochlea, optic nerve, and / or olfactory nerves. As used herein, the term CNS cells refer to any cells of the CNS including neurons and glial cells (oligodendrocytes, astrocytes, ependymal cells, microglia). The PNS refers to the nerves and ganglia outside the brain and spinal cord.

[0057] Depending on the type of disease, the target tissue or organ may comprise essentially the nervous system (CNS and / or PNS), essentially muscles or both the nervous system and muscles.

[0058] In some embodiments, the gene of interest is a functional version of a gene or a fragment thereof. The functional version of said gene includes the wild-type gene, a variant gene such as variants belonging to the same family and others, or a truncated version, which preserves the functionality of the encoded protein at least partially. A functional version of a gene is useful for replacement or additive gene therapy to replace a gene, which is deficient or non-functional in a patient. For example, a functional version of the dystrophin gene includes the various mini-dystrophin and micro-dystrophin gene constructs that are well- known in the art and other functional versions of the dystrophin gene. In other embodiments, the gene of interest is a gene which inactivates a dominant allele causing an autosomal dominant genetic disease. A fragment of a gene is useful as recombination template for use in combination with a genome editing enzyme.

[0059] Alternatively, the gene of interest may encode a protein of interest for a particular application, (for example an antibody or antibody fragment, a genome-editing enzyme) or a RNA. In some embodiments, the protein is a therapeutic protein including a therapeutic antibody or antibody fragment, or a genome-editing enzyme. In some embodiments, the RNA is a therapeutic RNA.

[0060] In some embodiments, the sequence of the gene of interest is optimized for expression in the treated individual, preferably a human individual. Sequence optimization may include a number of changes in a nucleic acid sequence, including codon optimization, increase of GC content, decrease of the number of CpG islands, decrease of the number of alternative open reading frames (ARFs) and / or decrease of the number of splice donor and splice acceptor sites.

[0061] The gene of interest is a functional gene able to produce the encoded protein, peptide or RNA in the target cells of the disease. In some embodiments, the gene of interest is a human gene. The AAV vectors comprise the gene of interest in a form expressible in cells of target organs. In particular, the gene of interest is operably linked to appropriate regulatory sequences for expression of a transgene in the individual’s target cells, tissue(s) or organ(s).Such sequences which are well-known in the art include in particular a promoter, and further regulatory sequences capable of further controlling the expression of a transgene, such as without limitation, enhancer, terminator, intron, silencer, in particular tissue-specific silencer, and microRNA. The gene of interest is operably linked to a ubiquitous, tissue-specific or inducible promoter which is functional in cells of target organs. In some particular embodiments, the gene of interest is operably linked to at least two promoters, wherein at least one of them is target cells specific- or inducible promoter which is functional in cells of target tissue or organ cells. In some particular embodiments, the gene of interest is operably linked to at least two promoters wherein one of them is target cells specific- or inducible promoter which is functional in cells of a target tissue or organ and the other is target-specific or inducible promoter which is functional in cells of another target tissue or organ which is different from the first one. The gene of interest may be inserted in an expression cassette further comprising additional regulatory sequences as disclosed above.

[0062] Examples of ubiquitous promoters include the CAG promoter, phosphoglycerate kinase 1 (PGK) promoter, the cytomegalovirus enhancer / promoter (CMV), the SV40 early promoter, the retroviral Rous sarcoma virus (RSV) LTR promoter, the dihydrofolate reductase promoter, the β-actin promoter, and the EF1 promoter.

[0063] Muscle-specific promoters include without limitation, the desmin (Des) promoter, muscle creatine kinase (MCK) promoter, alpha-myosin heavy chain (alpha-MHC) promoter, myosin light chain 2 (MLC-2) promoter, cardiac troponin C (cTnC) promoter, human skeletal actin (HSA) promoter or synthetic muscle-specific promoter such as SpC5-12 promoter, CK6 promoter, or MHCK7 promoter.

[0064] Promoters for nervous system, such as CNS expression include promoters driving ubiquitous expression and promoters driving expression into neurons. Representative promoters driving ubiquitous expression, without limitation: CAG promoter (includes the cytomegalovirus enhancer / chicken beta actin promoter, the first exon and the first intron of the chicken beta-actin gene and the splice acceptor of the rabbit beta-globin gene) ; PGK (phosphoglycerate kinase 1) promoter ; β-actin promoter ; EF1a promoter ; CMV promoter. Representative promoters driving expression into neurons include, without limitation, the promoter of the Calcitonin Gene-Related Peptide (CGRP), a known motor neuron-derived factor. Other neuron-selective promoters include the promoters of Choline Acetyl Transferase(ChAT), Neuron Specific Enolase (NSE), Synapsin, Hb9 and ubiquitous promoters including Neuron-Restrictive Silencer Elements (NRSE). Representative promoters driving selective expression in glial cells include the promoter of the Glial Fibrillary Acidic Protein gene (GFAP).

[0065] For expression in muscle cells (skeletal and cardiac muscle cells), the gene of interest is advantageously under the control of a desmin promoter, in particular human desmin promoter (Raguz et al., Dev. Biol., 1998, 201, 26-42; Paulin D & Li Z, Exp. Cell. Res., 2004, Nov 15;301(1):1-7) or MHCK7 promoter. For expression in skeletal muscle cells, the gene of interest is advantageously under the control of a desmin promoter, in particular human desmin promoter, and further comprises a miR208a target sequence that represses expression in cardiac muscle cells (i.e. in the heart; Roudault et al., Circulation, 2013, 128, 1094-104. doi: 10.1161 / CIRCULATIONAHA.113.001340).

[0066] The RNA is advantageously complementary to a target DNA or RNA sequence or binds to a target protein. For example, the RNA is an interfering RNA such as a shRNA, a microRNA, a guide RNA (gRNA) for use in combination with a Cas enzyme or similar enzyme for genome editing, an antisense RNA capable of exon skipping such as a modified small nuclear RNA (snRNA) or a long non-coding RNA. The interfering RNA or microRNA may be used to regulate the expression of a target gene involved in muscle and / or nervous system disease. The guide RNA in complex with a Cas enzyme or similar enzyme for genome editing may be used to modify the sequence of a target gene, in particular to correct the sequence of a mutated / deficient gene or to modify the expression of a target gene involved in a disease, in particular a muscle and / or nervous system disorder. The antisense RNA capable of exon skipping is used in particular to correct a reading frame and restore expression of a deficient gene having a disrupted reading frame. In some embodiments, the RNA is a therapeutic RNA.

[0067] The genome-editing enzyme according to the invention is any enzyme or enzyme complex capable of modifying a target gene or target cellular pathway. For example, the genome-editing enzyme may modify the expression, sequence or regulation of the target gene or cellular pathway. The genome-editing enzyme is advantageously an engineered nuclease, such as with no limitations, a meganuclease, zinc finger nuclease (ZFN), transcription activator-like effector-based nuclease (TALENs), Cas enzyme from clustered regularlyinterspaced palindromic repeats (CRISPR)-Cas system and similar enzymes. The genome- editing enzyme, in particular an engineered nuclease such as Cas enzyme and similar enzymes, may be a functional nuclease which generates a double-strand break (DSB) or single-stranded DNA break (nickase such as Cas9(D10A) in the target genomic locus and is used for site- specific genome editing applications, including with no limitations: gene correction, gene replacement, gene knock-in, gene knock-out, mutagenesis, chromosome translocation, chromosome deletion, and the like. For site-specific genome editing applications, the genome- editing enzyme, in particular an engineered nuclease such as Cas enzyme and similar enzymes may be used in combination with a homologous recombination (HR) matrix or template (also named DNA donor template) which modifies the target genomic locus by double-strand break (DSB)-induced homologous recombination. In particular, the HR template may introduce a transgene of interest into the target genomic locus or repair a mutation in the target genomic locus, preferably in an abnormal or deficient gene causing a muscle and / or nervous system disorder. The genome-editing enzyme, such as Cas enzyme and similar enzyme may be a DNA base-editor such as cytosine base-editor and adenine base-editor or a prime-editor. Base- editors can install all four transition mutations while Prime-editor expand the scope of donor- free precise DNA editing to not only all transition and transversion mutations, but small insertion and deletion mutations as well. Collectively, DNA base-editing and prime-editing tools enable precise nucleotide substitutions in a programmable manner, without requiring a donor template. Alternatively, the genome-editing enzyme, such as Cas enzyme and similar enzymes may be engineered to become nuclease-deficient and used as DNA-binding protein for various genome engineering applications such as with no limitation: transcriptional activation, transcriptional repression, epigenome modification, genome imaging, DNA or RNA pull-down and the like.

[0068] The combination of AAV vector serotypes according to the present disclosure is used in gene therapy of diseases, in particular targeted gene therapy directed to target tissue or organ.

[0069] The combination of AAV vector serotypes according to the present disclosure is preferably used in the form of separate pharmaceutical compositions comprising a therapeutically effective amount of AAV vector particles of each serotype, preferably AAV vector particles packaging a therapeutic gene of interest according to the present disclosure.

[0070] As used herein “Gene therapy” refers to a treatment of an individual which involves delivery of nucleic acid of interest into an individual's cells for the purpose of treating a disease. Delivery of the nucleic acid is generally achieved using a delivery vehicle, also known as a vector. The combination of rAAV vector particles according to the present disclosure may be employed to deliver a gene to a patient's cells.

[0071] As used herein “Cell therapy” refers to a process wherein cells stably transduced by a combination of rAAV vector particles according to the present disclosure are delivered to the individual in need thereof by any appropriate mean such as for example by intravenous injection (infusion), or injection in the tissue of interest (implantation or transplantation). In particular embodiments, cell therapy comprises collecting cells from the individual, transducing the individual’s cells with the combination of rAAV vector particles according to the present disclosure, and administering the stably transduced cells back to the patient. As used herein “cell” refers to isolated cell, natural or artificial cellular aggregate, bioartificial cellular scaffold and bioartificial organ or tissue.

[0072] Gene therapy can be performed by gene transfer, gene editing, exon skipping, RNA-interference, trans-splicing or any other genetic modification of any coding or regulatory sequences in the cell, including those included in the nucleus, mitochondria or as commensal nucleic acid such as with no limitation viral sequences contained in cells.

[0073] The two main types of gene therapy are the following: - a therapy aiming to provide a functional replacement gene for a deficient / abnormal gene: this is replacement or additive gene therapy; - a therapy aiming at gene or genome editing: in such a case, the purpose is to provide to a cell the necessary tools to correct the sequence or modify the expression or regulation of a deficient / abnormal gene so that a functional gene is expressed or an abnormal gene is suppressed (inactivated): this is gene editing therapy.

[0074] In additive gene therapy, the gene of interest may be a functional version of a gene, which is deficient or mutated in a patient, as is the case for example in a genetic disease. In such a case, the gene of interest will restore the expression of a functional gene.

[0075] Gene or genome editing uses one or more gene(s) of interest, such as:(i) a gene encoding a therapeutic RNA as defined above such as an interfering RNA like a shRNA or a microRNA, a guide RNA (gRNA) for use in combination with a Cas enzyme or similar enzyme, or an antisense RNA capable of exon skipping such as a modified small nuclear RNA (snRNA); and (ii) a gene encoding a genome-editing enzyme as defined above such as an engineered nuclease like a meganuclease, zinc finger nuclease (ZFN), transcription activator-like effector-based nuclease (TALENs), Cas enzyme or similar enzymes; or a combination of such genes, and maybe also a fragment of a functional version of a gene for use as recombination template, as defined above.

[0076] Gene therapy is used for treating various inherited (genetic) or acquired diseases or disorders affecting the structure or function of target tissue or organ. The diseases may be caused by trauma, infection, degeneration, structural or metabolic defects, tumors, inflammatory or autoimmune disorders, stroke or other causes.

[0077] In some particular embodiments, the disease affects one or more of the following target tissue or organs: muscles including skeletal, smooth or cardiac muscles; nervous system including brain or spinal cord; the disease may affect muscles including skeletal or cardiac muscles; nervous system including brain or spinal cord; or both.

[0078] In some particular embodiments, the disease is a myopathy such as skeletal and / or cardiomyopathy, preferably a genetic myopathy.

[0079] In some particular embodiments, the disease is a neurological disorder (nervous system disease), preferably a genetic neurological disorder. The neurological disorder may affect the CNS and / or PNS such as the brain, spinal cord or both. CNS diseases include for example Alzheimer, Parkinson, Frontotemporal dementia, Huntington disease and others.

[0080] In some particular embodiments, the disease is a neuromuscular disorder, preferably a genetic neuromuscular disorder. Neuromuscular disease or disorder (NMD) is a very broad term encompassing a range of conditions that impair the functioning of the muscles, either directly, being pathologies of the voluntary muscle, or indirectly, being pathologies of the peripheral nervous system or neuromuscular junctions. Neuromuscular diseases are a broadly defined group of disorders that all involve injury or dysfunction of peripheral nerves or muscle or neuromuscular junctions. The site of injury can be in the cellbodies (i.e., amyotrophic lateral sclerosis [ALS] or sensory ganglionopathies), axons (i.e., axonal peripheral neuropathies or brachial plexopathies), Schwann cells (i.e., chronic inflammatory demyelinating polyradiculoneuropathy), neuromuscular junction (i.e., myasthenia gravis or Lambert-Eaton myasthenic syndrome), muscle (i.e., inflammatory myopathy or muscular dystrophy), or any combination of these sites. Some neuromuscular diseases are also associated with central nervous system disease, such as ALS or spinal muscular atrophies.

[0081] Examples of mutated genes in genetic neurological disorders, that can be targeted by gene therapy using the combination of AAV vectors of the invention are listed in the following table:

[0082] Genetic neurological disorders Disease Gene Friedreich ataxia FXN Ataxia-telangectasia ATM Niemann pick disease SMPD1, NPC1,2 Spinocerebellar ataxia ATXN1-3, PLEKHG4, SPTBN2, CACNA1A, ATXN 7 Marinesco-Sjoren SIL1 Juvenile Parkinsonism SNCA, PARK2, PINK1, GCH1, TH Dystonia TOR1A, GCH1 Huntington’s disease HTT Wilson disease ATP7B Pantothenate kinase- PANK2 associated neurodegeneration Lysosomal Storage Disorders GBA, SMPD1, NPC1, NPC2 (LSDs) Fragile X syndrome FMR1 Rett Syndrome MECP2 Adrenoleukodystrophy ABCD1 Epilepsy BFNS, KCNQ2, KCNQ3, BFNIS, SCN2A, BFIS, PRRT2, CHRNA4, CHRNB2, CHRNA2, KCNT1, SCNA1, TBC1D24, SCN8A, CDKL5, ARX, STXBP1, SCN1B, GABRG2

[0083] Other examples of mutated genes in genetic neurological disorders, that can be targeted by gene therapy using the combination of AAV vectors of the invention are the genes responsible for Spinal muscular atrophies (SMAs) & Motor Neuron diseases; Hereditary motor and sensory neuropathies; Hereditary paraplegia and Hereditary ataxia; listed in thetables below. In some particular embodiments, said neurological disease is selected from the group consisting of: Spinal muscular atrophy (SMN1, ASAH1 genes), Amyotrophic lateral sclerosis (SOD1, ALS2, SETX, FUS, ANG, TARDBP, FIG4, OPTN and others); Hereditary paraplegia (SPAST (SPG4), SPG7, and other SPG genes such as SPG11, SPG20 and SPG21; in particular SPAST (SPG4) and SPG7) and Charcot-Marie-Tooth, Type 4B1 (MTMR2). In some preferred embodiments, said gene is selected from the group consisting of: SMN1, ASAH1, DNM2, MTMR2 and SPAST genes. In some other preferred embodiments, said gene is selected from the group consisting of: SOD1, ALS2, SETX, FUS, ANG, TARDBP, FIG4 and OPTN.

[0084] Examples of mutated genes in genetic myopathies that can be targeted by gene therapy using the combination of AAV vectors of the invention are listed in the following tables:

[0085] Muscular dystrophies Gene Protein DMD Dystrophin EMD Emerin FHL1 Four and a half LIM domain 1 LMNA Lamin A / C SYNE1 Spectrin repeat containing, nuclear envelope 1 (nesprin 1) SYNE2 Spectrin repeat containing, nuclear envelope 2 (nesprin 2) TMEM43 Transmembrane protein 43 TOR1AIP1 Torsin A interacting protein 1 DUX4 Double homeobox 4 SMCHD1 Structural maintenance of chromosomes flexible hinge domain containing 1 PTRF Polymerase I and transcript release factor MYOT Myotilin CAV3 Caveolin 3 DNAJB6 HSP-40 homologue, subfamily B, number 6 DES Desmin TNPO3 Transportin 3 HNRNPDL Heterogeneous nuclear ribonucleoprotein D-like CAPN3 Calpain 3 DYSF Dysferlin SGCG Gamma sarcoglycan SGCA Alpha sarcoglycanSGCB Beta sarcoglycan SGCD Delta-sarcoglycan TCAP Telethonin TRIM32 Tripartite motif-containing 32 FKRP Fukutin-related protein TTN Titin POMT1 Protein-O-mannosyltransferase 1 ANO5 Anoctamin 5 FKTN Fukutin POMT2 Protein-O-mannosyltransferase 2 POMGNT1 O-linked mannose beta1,2-N-acetylglucosaminyltransferase PLEC Plectin TRAPPC11 trafficking protein particle complex 11 GMPPB GDP-mannose pyrophosphorylase B DAG1 Dystroglycan1 DPM3 Dolichyl-phosphate mannosyltransferase polypeptide 3 ISPD Isoprenoid synthase domain containing VCP Valosin-containing protein LIMS2 LIM and senescent cell antigen-like domains 2 GAA Glucosidase alpha, acid

[0086] Congenital muscular dystrophies Gene Protein LAMA2 Laminin alpha 2 chain of merosin COL6A1 Alpha 1 type VI collagen COL6A2 Alpha 2 type VI collagen COL6A3 Alpha 3 type VI collagen SEPN1 Selenoprotein N1 FHL1 Four and a half LIM domain 1 ITGA7 Integrin alpha 7 precursor DNM2 Dynamin 2 TCAP Telethonin LMNA Lamin A / C FKTN Fukutin POMT1 Protein-O-mannosyltransferase 1 POMT2 Protein-O-mannosyltransferase 2 FKRP Fukutin-related protein POMGNT1 O-linked mannose beta1,2-N-acetylglucosaminyltransferase ISPD Isoprenoid synthase domain containing POMGNT2 protein O-linked mannose N-acetylglucosaminyltransferase 2B3GNT1 UDP-GlcNAc:betaGal beta-1,3-N-acetylglucosaminyl-transferase 1 GMPPB GDP-mannose pyrophosphorylase B LARGE Like-glycosyltransferase DPM1 Dolichyl-phosphate mannosyltransferase 1, catalytic subunit DPM2 Dolichyl-phosphate mannosyltransferase polypeptide 2, regulatory subunit ALG13 UDP-N-acetylglucosami-nyltransferase B3GALNT2 Beta-1,3-N-acetylgalacto-saminyltransferase 2 TMEM5 Transmembrane protein 5 POMK Protein-O-mannose kinase CHKB Choline kinase beta ACTA1 Alpha actin, skeletal muscle TRAPPC11 trafficking protein particle complex 11

[0087] Congenital myopathies Gene Protein TPM3 Tropomyosin 3 NEB Nebulin ACTA1 Alpha actin, skeletal muscle TPM2 Tropomyosin 2 (beta) TNNT1 Slow troponin T KBTBD13 Kelch repeat and BTB (POZ) domain containing 13 CFL2 Cofilin 2 (muscle) KLHL40 Kelch-like family member 40 KLHL41 Kelch-like family member 41 LMOD3 Leiomodin 3 (fetal) SEPN1 Selenoprotein N1 RYR1 Ryanodine receptor 1 (skeletal) MYH7 Myosin, heavy polypeptide 7, cardiac muscle, beta MTM1 Myotubularin DNM2 Dynamin 2 BIN1 Amphiphysin TTN Titin SPEG SPEG complex locus MEGF10 Multiple EGF-like-domains 10 MYH2 Myosin, heavy polypeptide 2, skeletal muscle MYBPC3 Cardiac myosin binding protein-C CNTN1 Contactin-1 TRIM32 Tripartite motif-containing 32 PTPLA Protein tyrosine phosphatase-like (3-Hydroxyacyl-CoA dehydratase CACNA1S Calcium channel, voltage-dependent, L type, alpha 1S subunit

[0088] Distal myopathies Gene symbol protein DYSF Dysferlin TTN Titin GNE UDP-N-acetylglucosamine-2- epimerase / N-acetylmannosamine kinase MYH7 Myosin, heavy polypeptide 7, cardiac muscle, beta MATR3 Matrin 3 TIA1 Cytotoxic granuleassociated RNA binding protein MYOT Myotilin NEB Nebulin CAV3 Caveolin 3 LDB3 LIM domain binding 3 ANO5 Anoctamin 5 DNM2 Dynamin 2 KLHL9 Kelch-like homologue 9 FLNC Filamin C, gamma (actin-binding protein - 280) VCP Valosin-containing protein

[0089] Other myopathies Gene symbol protein ISCU Iron-sulfur cluster scaffold homolog (E. coli) MSTN Myostatin FHL1 Four and a half LIM domain 1 BAG3 BCL2-associated athanogene 3 ACVR1 Activin A receptor, type II-like kinase 2 MYOT Myotilin FLNC Filamin C, gamma (actin-binding protein - 280) LDB3 LIM domain binding 3 LAMP2 Lysosomal-associated membrane protein 2 precursor VCP Valosin-containing protein CAV3 Caveolin 3 SEPN1 Selenoprotein N1 CRYAB Crystallin, alpha B DES Desmin VMA21 VMA21 Vacuolar H+-ATPase Homolog (S. Cerevisiae) PLEC plectin PABPN1 Poly(A) binding protein, nuclear 1 TTN Titin RYR1 Ryanodine receptor 1 (skeletal)CLN3 Ceroid-lipofuscinosis, neuronal 3 (=battenin) TRIM54 TRIM63 Tripartite motif containing 63, E3 ubiquitin protein ligase

[0090] Myotonic syndromes Gene protein DMPK Myotonic dystrophy protein kinase CNBP (ZNF9) Cellular nucleic acid-binding protein CLCN1 Chloride channel 1, skeletal muscle (Thomsen disease, autosomal dominant) CAV3 Caveolin 3 HSPG2 Perlecan ATP2A1 ATPase, Ca++ transporting, fast twitch 1

[0091] Ion Channel muscle diseases Gene protein CLCN1 Chloride channel 1, skeletal muscle (Thomsen disease, autosomal dominant) SCN4A Sodium channel, voltage-gated, type IV, alpha SCN5A Voltage-gated sodium channel type V alpha CACNA1S Calcium channel, voltage-dependent, L type, alpha 1S subunit CACNA1A Calcium channel, voltage-dependent, P / Q type, alpha 1A subunit KCNE3 Potassium voltage-gated channel, Isk-related family, member 3 KCNA1 Potassium voltage-gated channel, shaker-related subfamily, member 1 KCNJ18 Kir2.6 (inwardly rectifying potassium channel 2.6) KCNJ2 Potassium inwardly-rectifying channel J2 KCNH2 Voltage-gated potassium channel, subfamily H, member 2 KCNQ1 Potassium voltage-gated channel, KQT-like subfamily, member 1 KCNE2 Potassium voltage-gated channel, Isk-related family, member 2 KCNE1 Potassium voltage-gated channel, Isk-related family, member 1

[0092] Malignant hyperthermia Gene protein RYR1 Ryanodine receptor 1 (skeletal) CACNA1S Calcium channel, voltage-dependent, L type, alpha 1S subunit

[0093] Metabolic myopathies Gene protein GAA Acid alpha-glucosidase preproprotein AGL Amylo-1,6-glucosidase, 4-alpha-glucanotransferase GBE1 Glucan (1,4-alpha-), branching enzyme 1 (glycogen branching enzyme, Andersen disease, glycogen storage disease type IV) PYGM Glycogen phosphorylase PFKM Phosphofructokinase, muscle PHKA1 Phosphorylase b kinase, alpha submit PGM1 Phosphoglucomutase 1 GYG1 Glycogenin 1 GYS1 Glycogen synthase 3 glycogen synthase 1 (muscle) glycogen synthase 1 (muscle) PRKAG2 Protein kinase, AMP-activated, gamma 2 non-catalytic subunit RBCK1 RanBP-type and C3HC4-type zinc finger containing 1 (heme-oxidized IRP2 ubiquitin ligase 1) PGK1 Phosphoglycerate kinase 1 PGAM2 Phosphoglycerate mutase 2 (muscle) LDHA Lactate dehydrogenase A ENO3 Enolase 3, beta muscle specific CPT2 Carnitine palmitoyltransferase II SLC22A5 Solute carrier family 22 member 5 SLC25A20 Carnitine-acylcarnitine translocase ETFA Electron-transfer-flavoprotein, alpha polypeptide ETFB Electron-transfer-flavoprotein, beta polypeptide ETFDH Electron-transferring-flavoprotein dehydrogenase ACADVL Acyl-Coenzyme A dehydrogenase, very long chain ABHD5 Abhydrolase domain containing 5 PNPLA2 Adipose triglyceride lipase (desnutrin) LPIN1 Lipin 1 (phosphatidic acid phosphatase 1) PNPLA8 Patatin-like phospholipase domain containing 8

[0094] Hereditary Cardiomyopathies Gene protein MYH6 Myosin heavy chain 6 MYH7 Myosin, heavy polypeptide 7, cardiac muscle, beta TNNT2 Troponin T2, cardiac TPM1 Tropomyosin 1 (alpha) MYBPC3 Cardiac myosin binding protein-C PRKAG2 Protein kinase, AMP-activated, gamma 2 non-catalytic subunit TNNI3 Troponin I, cardiacMYL3 Myosin light chain 3 TTN Titin MYL2 Myosin light chain 2 ACTC1 Actin, alpha, cardiac muscle precursor CSRP3 Cysteine and glycine-rich protein 3 (cardiac LIM protein) TNNC1 Slow troponin C VCL Vinculin MYLK2 Myosin light chain kinase 2 CAV3 Caveolin 3 MYOZ2 Myozenin 2, or calsarcin 1, a Z disk protein JPH2 Junctophilin-2 PLN Phospholamban NEXN Nexilin(F-actin binding protein) ANKRD1 Ankyrin repeat domain 1 (cardiac muscle) ACTN2 Actinin alpha2 NDUFAF1 NADH-ubiquinone oxidoreductase 1 alpha subcomplex TSFM Ts translation elongation factor, mitochondrial AARS2 Alanyl-tRNA synthetase 2, mitochondrial MRPL3 Mitochondrial ribosomal protein L3 COX15 COX15 homolog, cytochrome c oxidase assembly protein (yeast) MTO1 Mitochondrial tRNA translation optimization 1 MRPL44 Mitochondrial ribosomal protein L44 LMNA Lamin A / C LDB3 LIM domain binding 3 SCN5A Voltage-gated sodium channel type V alpha DES Desmin EYA4 Eyes absent 4 SGCD Delta-sarcoglycan TCAP Telethonin ABCC9 ATP-binding cassette, sub-family C (member 9) TMPO Lamina-associated polypeptide 2 PSEN2 Presenilin 2 CRYAB Crystallin, alpha B FKTN Fukutin TAZ Tafazzin DMD Dystrophin LAMA4 Laminin alpha 4 ILK Integrin-linked kinase MYPN Myopalladin RBM20 RNA binding motif protein 20 SYNE1 Spectrin repeat containing, nuclear envelope 1 (nesprin 1)MURC Muscle-related coiled-coil protein DOLK Dolichol kinase GATAD1 GATA zinc finger domain containing 1 SDHA succinate dehydrogenase complex, subunit A, flavoprotein (Fp) GAA Acid alpha-glucosidase preproprotein DTNA Dystrobrevin, alpha FLNA Filamin A, alpha (actin binding protein 280) TGFB3 Transforming growth factor, beta 3 RYR2 Ryanodine receptor 2 TMEM43 Transmembrane protein 43 DSP Desmoplakin PKP2 Plakophilin 2 DSG2 Desmoglein 2 DSC2 Desmocollin 2 JUP Junction plakoglobin CASQ2 Calsequestrin 2 (cardiac muscle) KCNQ1 Potassium voltage-gated channel, KQT-like subfamily, member 1 KCNH2 Voltage-gated potassium channel, subfamily H, member 2 ANK2 Ankyrin 2 KCNE1 Potassium voltage-gated channel, Isk-related family, member 1 KCNE2 Potassium voltage-gated channel, Isk-related family, member 2 KCNJ2 Potassium inwardly-rectifying channel J2 CACNA1C Calcium channel, voltage-dependent, L type, alpha 1C subunit SCN4B Sodium channel, voltage-gated, type IV, beta subunit AKAP9 A kinase (PRKA) anchor protein (yotiao) 9 SNTA1 Syntrophin, alpha 1 KCNJ5 Potassium inwardly-rectifying channel, subfamily J, member 5 NPPA Natriuretic peptide precursor A KCNA5 Potassium voltage-gated channel, shaker-related subfamily, member 5 GJA5 Connexin 40 SCN1B Sodium channel, voltage-gated, type I, beta subunit SCN2B Sodium channel, voltage-gated, type II, beta subunit NUP155 Nucleoporin 155 kDa GPD1L Glycerol-3-phosphate dehydrogenase 1-like CACNB2 Calcium channel, voltage-dependent, beta 2 subunit KCNE3 Potassium voltage-gated channel, Isk-related family, member 3 SCN3B Sodium channel, voltage-gated, type III, beta subunit HCN4 Hyperpolarization activated cyclic nucleotide-gated potassium channel 4

[0095] Congenital myasthenic syndromes Gene protein CHRNA1 Cholinergic receptor, nicotinic, alpha polypeptide 1 CHRNB1 Cholinergic receptor, nicotinic, beta 1 muscle CHRND Cholinergic receptor, nicotinic, delta CHRNE Cholinergic receptor, nicotinic, epsilon RAPSN Rapsyn CHAT Choline acetyltransferase isoform COLQ Acetylcholinesterase collagen-like tail subunit MUSK muscle, skeletal, receptor tyrosine kinase DOK7 Docking protein 7 AGRN Agrin GFPT1 Glutamine-fructose-6-phosphate transaminase 1 DPAGT1 Dolichyl-phosphate (UDP-N-acetylglucosamine) N- acetylglucosaminephosphotransferase 1 (GlcNAc-1-P transferase) LAMB2 Laminin, beta 2 (laminin S) SCN4A Sodium channel, voltage-gated, type IV, alpha CHRNG Cholinergic receptor, nicotinic, gamma polypeptide PLEC plectin ALG2 Alpha-1,3 / 1,6-mannosyltransferase ALG14 UDP-N-acetylglucosaminyltransferase SYT2 Synaptotagmin II PREPL Prolyl endopeptidase-like

[0096] Spinal muscular atrophies (SMAs) &Motor Neuron diseases Gene protein SMN1 Survival of motor neuron 1, telomeric IGHMBP2 Immunoglobulin mu binding protein 2 PLEKHG5 Pleckstrin homology domain containing, family G (with RhoGef domain) member 5 HSPB8 Heat shock 27kDa protein 8 HSPB1 Heat shock 27kDa protein 1 HSPB3 Heat shock 27kDa protein 3 AARS Alanyl-tRNA synthetase GARS Glycyl-tRNA synthetase BSCL2 Seipin REEP1 Receptor accessory protein 1 SLC5A7 Solute carrier family 5 (sodium / choline cotransporter), member 7 DCTN1 Dynactin 1 UBA1 Ubiquitin-activating enzyme 1 ATP7A ATPase, Cu++ transporting, alpha polypeptideDNAJB2 DnaJ (Hsp40) homolog, subfamily B, member 2 TRPV4 Transient receptor potential cation channel, subfamily V, member 4 DYNC1H1 Dynein, cytoplasmic 1, heavy chain 1 BICD2 Bicaudal D homolog 2 (Drosophila) FBXO38 F-box protein 38 ASAH1 N-acylsphingosine amidohydrolase (acid ceramidase) 1 VAPB Vesicle-associated membrane protein-associated protein B and C EXOSC8 Exosome component 8 SOD1 Superoxide dismutase 1, soluble ALS2 Alsin SETX Senataxin FUS Fusion (involved in t(12;16) in malignant liposarcoma) ANG Angiogenin TARDBP TAR DNA binding protein FIG4 Sac domain-containing inositol phosphatase 3 OPTN Optineurin ATXN2 Ataxin 2 VCP Valosin-containing protein UBQLN2 Ubiquilin 2 SIGMAR1 Sigma non-opioid intracellular receptor 1 CHMP2B Charged multivesicular body protein 2B PFN1 Profilin 1 MATR3 Matrin 3 NEFH Neurofilament, heavy polypeptide PRPH Peripherin C9orf72 Chromosome 9 open reading frame 72 CHCHD10 Coiled-coil-helix-coiled-coil-helix domain containing 10 SQSTM1 Sequestosome 1 AR Androgen receptor GLE1 GLE1 RNA export mediator homolog (yeast) ERBB3 V-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (avian) PIP5K1C Phosphatidylinositol-4-phosphate 5-kinase, type I, gamma EXOSC3 Exosome component 3 VRK1 Vaccinia related kinase 1 SLC52A3 Solute carrier family 52, riboflavin transporter, member 3 SLC52A2 Solute carrier family 52, riboflavin transporter, member 2 HEXB Hexosaminidase B

[0097] Hereditary motor and sensory neuropathies Gene Protein PMP22 Peripheral myelin protein 22 MPZ Myelin protein zero LITAF Lipopolysaccharide-induced TNF factor EGR2 Early growth response 2 protein NEFL Neurofilament, light polypeptide 68kDa HOXD10 Homeobox D10 ARHGEF10 Rho guanine nucleotide exchange factor 10 FBLN5 Fibulin 5 (extra-cellular matrix) DNM2 Dynamin 2 YARS Tyrosyl-tRNA synthetase INF2 Inverted formin 2 GNB4 Guanine nucleotidebinding protein, beta-4 GDAP1 Ganglioside-induced differentiation-associated protein 1 MTMR2 Myotubularin-related protein 2 SBF2 SET binding factor 2 SBF1 SET binding factor 1 SH3TC2 KIAA1985 protein NDRG1 N-myc downstream regulated gene 1 PRX Periaxin HK1 Hexokinase 1 FGD4 Actin-filament binding protein Frabin FIG4 Sac domain-containing inositol phosphatase 3 SURF1 surfeit 1 GJB1 Gap junction protein, beta 1, 32kDa (connexin 32) AIFM1 Apoptosis-inducing factor, mitochondrionassociated 1 PRPS1 Phosphoribosyl pyrophosphate synthetase 1 PDK3 Pyruvate dehydrogenase kinase, isoenzyme 3 KIF1B Kinesin family member 1B MFN2 Mitofusin 2 RAB7A RAB7, member RAS oncogene family TRPV4 Transient receptor potential cation channel, subfamily V, member 4 GARS Glycyl-tRNA synthetase HSPB1 Heat shock 27kDa protein 1 HSPB8 Heat shock 27kDa protein 8 AARS Alanyl-tRNA synthetase DYNC1H1 Dynein, cytoplasmic 1, heavy chain 1 LRSAM1 leucine rich repeat and sterile alpha motif containing 1 DHTKD1 dehydrogenase E1 and transketolase domain containing 1TRIM2 Tripartite motif containing 2 TFG TRK-fused gene MARS methionyl-tRNA synthetase KIF5A Kinesin family member 5A LMNA Lamin A / C MED25 Mediator complex subunit 25 DNAJB2 DnaJ (Hsp40) homolog, subfamily B, member 2 HINT1 Histidine triad nucleotide binding protein 1 KARS Lysyl-tRNA synthetase PLEKHG5 Pleckstrin homology domain containing, family G (with RhoGef domain) member 5 COX6A1 Cytochrome c oxidase subunit VIa polypeptide 1 IGHMBP2 Immunoglobulin mu binding protein 2 SPTLC1 Serine palmitoyltransferase subunit 1 SPTLC2 Serine palmitoyltransferase long chain base subunit 2 ATL1 Atlastin GTPase 1 KIF1A Kinesin family member 1A WNK1 WNK lysine deficient protein kinase 1 IKBKAP Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase complex-associated protein NGF Nerve growth factor (beta polypeptide) DNMT1 DNA (cytosine-5)-methyltransferase 1 SLC12A6 Potassium chloride cotransporter KCC3 GJB3 Gap junction protein, beta 3, 31kDa (=connexin 31) sept-09Septin 9GAN Gigaxonin CTDP1 CTD phosphatase subunit 1 VRK1 Vaccinia related kinase 1

[0098] Hereditary paraplegia Gene symbol protein ATL1 Atlastin SPAST Spastin NIPA1 Non-imprinted in Prader-Willi / Angelman syndrome 1 KIAA0196 Strumpellin KIF5A Kinesin family member 5A RTN2 Reticulon 2 HSPD1 Heat shock 60kDa protein 1 (chaperonin) BSCL2 Seipin REEP1 Receptor accessory protein 1 ZFYVE27 ProtrudinSLC33A1 Solute carrier family 33 (acetyl- CoA transporter) CYP7B1 Cytochrome P450, family 7, subfamily B, polypeptide 1 SPG7 Paraplegin SPG11 Spatacsin ZFYVE26 Spastizin ERLIN2 ER lipid raft associated 2 SPG20 Spartin SPG21 Maspardin B4GALNT1 beta-1,4-N-acetyl-galactosaminyl transferase 1 DDHD1 DDHD domain containing 1 KIF1A Kinesin family member 1A FA2H Fatty acid 2-hydroxylase PNPLA6 Patatin-like phospholipase domain containing 6 C19orf12 chromosome 19 open reading frame 12 GJC2 gap junction protein, gamma 2, 47kDa NT5C2 5'-nucleotidase, cytosolic II GBA2 glucosidase, beta (bile acid) 2 AP4B1 adaptor-related protein complex 4, beta 1 subunit AP5Z1 Hypothetical protein LOC9907 TECPR2 tectonin beta-propeller repeat containing 2 AP4M1 Adaptor-related protein complex 4, mu 1 subunit AP4E1 Adaptor-related protein complex 5, zeta 1 subunit AP4S1 adaptor-related protein complex 4, sigma 1 subunit DDHD2 DDHD domain containing 2 C12orf65 adaptor-related protein complex 4, sigma 1 subunit CYP2U1 cytochrome P450, family 2, subfamily U, polypeptide 1 ARL6IP1 ADP-ribosylation factor-like 6 interacting protein 1 AMPD2 adenosine monophosphate deaminase 2 ENTPD1 ectonucleoside triphosphate diphosphohydrolase 1 ALDH3A2 Aldehyde dehydrogenase 3A2 ALS2 Alsin L1CAM L1 cell adhesion molecule PLP1 Proteolipid protein 1 MTPAP mitochondrial poly(A) polymerase AFG3L2 AFG3 ATPase family gene 3-like 2 (S. cerevisiae) 1 SACS Sacsin

[0099] Other neuromuscular disorders Gene protein TOR1A Torsin A SGCE Sarcoglycan, epsilon IKBKAP Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase complex-associated protein TTR Transthyretin (prealbumin, amyloidosis type I) KIF21A Kinesin family member 21A PHOX2A Paired-like aristaless homeobox protein 2A TUBB3 Tubulin, beta 3 TPM2 Tropomyosin 2 (beta) MYH3 Myosine, heavy chain 3, skeletal muscle, embryonic TNNI2 Troponin I, type 2 TNNT3 Troponin T3, skeletal SYNE1 Spectrin repeat containing, nuclear envelope 1 (nesprin 1) MYH8 Myosin heavy chain, 8, skeletal muscle, perinatal POLG Polymerase (DNA directed), gamma SLC25A4 Mitochondrial carrier; adenine nucleotide translocator C10orf2 chromosome 10 open reading frame 2 POLG2 Mitochondrial DNA polymerase, accessory subunit RRM2B Ribonucleotide reductase M2 B (TP53 inducible) TK2 Thymidine kinase 2, mitochondrial SUCLA2 Succinate-CoA ligase, ADP-forming, beta subunit OPA1 optic atrophy 1 STIM1 Stromal interaction molecule 1 ORAI1 ORAI calcium release-activated calcium modulator 1 PUS1 Pseudouridylate synthase 1 CHCHD10 Coiled-coil-helix-coiled-coil-helix domain containing 10 CASQ1 Calsequestrin 1 (fast-twitch, skeletal muscle) YARS2 tyrosyl-tRNA synthetase 2, mitochondrial [000100] Hereditary ataxia Gene protein symbol ATXN1 Ataxin 1 ATXN2 Ataxin 2 ATXN3 Ataxin 3 SPTBN2 Spectrin, beta, non-erythrocytic 2 CACNA1 Calcium channel, voltage-dependent, P / Q type, alpha 1A subunit A ATXN7 Ataxin 7ATXN8OS Ataxin 8 opposite strand ATXN10 Ataxin 10 TTBK2 Tau tubulin kinase 2 PPP2R2B Protein phosphatase 2 regulatory subunit B, beta isoform KCNC3 Potassium voltage-gated channel, Shaw-related subfamily, member 3 PRKCG Protein kinase C, gamma ITPR1 Inositol 1,4,5-triphosphate receptor type 1 TBP TATA box binding protein IFRD1 Interferon-related developmental regulator 1 KCND3 Potassium voltage-gated channel, Shal-related subfamily, member 3 PDYN prodynorphin EEF2 Eukaryotic translation elongation factor 2 FGF14 Fibroblast growth factor 14 AFG3L2 AFG3 ATPase family gene 3-like 2 (S. cerevisiae) 1 BEAN1 Brain expressed, associated with Nedd42 TK2 Thymidine kinase 2, mitochondrial ELOVL4 ELOVL fatty acid elongase 4 TGM6 Transglutaminase 6 NOP56 NOP56 ribonucleoprotein ELOVL5 ELOVL fatty acid elongase 5 CCDC88C Coiled-coil domain containing 88C KCNA1 Potassium voltage-gated channel, shaker-related subfamily, member 1 CACNB4 Calcium channel, voltage-dependent, beta 4 subunit SLC1A3 EAAT1 (excitatory amino acid transporter type 1) FXN Frataxin TTPA Tocopherol (alpha) transfer protein (ataxia (Friedreich-like) with vitamin E deficiency) C10orf2 chromosome 10 open reading frame 2 APTX Aprataxin SETX Senataxin SYNE1 Spectrin repeat containing, nuclear envelope 1 (nesprin 1) ADCK3 Atypical kinase ADCK3, mitochondrial TDP1 Tyrosyl-DNA phosphodiesterase 1 SIL1 SIL1 homolog, endoplasmic reticulum chaperone POLG Polymerase (DNA directed), gamma ATM Ataxia telangiectasia mutated MRE11A MRE11 meiotic recombination 11 homolog A SACS Sacsin PHYH Phytanoyl-CoA 2-hydroxylase PEX7 Peroxisomal biogenesis factor 7 RNF216 Ring finger protein 216[000101] Any one of the above listed genes may be targeted in replacement gene therapy, wherein the gene of interest is a functional version of the deficient or mutated gene. [000102] Alternatively, the above listed genes may be used as target for gene editing. Gene editing is used to correct the sequence of a mutated gene or modify the expression or regulation of a deficient / abnormal gene so that a functional gene is expressed in muscle cells. In such cases, the gene of interest is chosen from those encoding therapeutic RNAs such as interfering RNAs, guide RNAs for genome editing and antisense RNAs capable of exon skipping, wherein the therapeutic RNAs target the preceding list of genes. Tools such as CRISPR / Cas9 may be used for that purpose. [000103] Thus, by gene editing or gene replacement a correct version of this gene is provided in target cells of affected patients, in particular muscle cells and / or cells of the nervous system (PNS and / or CNS) of affected patients, this may contribute to effective therapies against this disease. [000104] In some embodiments, the target gene for gene therapy (additive gene therapy or gene editing) is a gene responsible for a neurological disorder, a muscular disorder, or a neuromuscular disorder, as disclosed herein. [000105] Disease that can treated by gene therapy using a combination of AAV vector serotypes according to the disclosure include in particular: Muscular dystrophies, Congenital muscular dystrophies, Congenital myopathies, Distal myopathies, Other myopathies, Myotonic syndromes, Ion Channel muscle diseases, Malignant hyperthermia, Metabolic myopathies, Hereditary Cardiomyopathies, Congenital myasthenic syndromes, Myastenia, Spinal muscular atrophies (SMAs) and Motor Neuron diseases, Hereditary paraplegia, Hereditary ataxia, Hereditary motor and sensory neuropathies and other neuromuscular disorders; the disease can be treating by targeting the gene associated with said diseases as listed in the Tables above. These diseases may be classified in different groups: (i) Myopathies include hereditary cardiomyopathies, metabolic myopathies, other myopathies, distal myopathies, muscular dystrophies and congenital myopathies; muscular dystrophies include Duchenne muscular dystrophies; congenital myopathies include myotubular myopathy and centronuclear myopathies (ii) Spinal muscular atrophies (SMAs) and motor neuron diseases include amyotrophic lateral sclerosis (ALS), progressive bulbar palsy (PBP), pseudobulbarpalsy, progressive muscular atrophy (PMA), primary lateral sclerosis (PLS), spinal muscular atrophy (SMA) such as spinal muscular atrophy with progressive myoclonic epilepsy (SMA- PME), SMN1-related spinal muscular atrophy, spinobulbar muscular atrophy (SBMA) and monomelic atrophy (MMA), as well as some rarer variants resembling ALS; (iii) Myotonic syndrome includes myotonic dystrophy type 1 (DMPK gene) and type 2 (CNBP gene); (iv) Congenital myasthenic syndromes and Myastenia; (v) Hereditary motor and sensory neuropathies ; (vi) Hereditary paraplegia and Hereditary ataxia. [000106] In some preferred embodiments, the genetic neuromuscular disease is selected from the group comprising : (i) myopathies, such as congenital myopathies, myasthenic syndromes, metabolic myopathies, distal myopathies, muscular dystrophies with or without cardiomyopathy; and (ii) spinal muscular atrophies (SMAs) and motor neuron diseases; preferably SMN1-related spinal muscular atrophy, spinal muscular atrophy with progressive myoclonic epilepsy (SMA-PME), spinobulbar muscular atrophy (SBMA) and amyotrophic lateral sclerosis (ALS). [000107] Examples of neuromuscular genetic disorders that can be treated using a combination of AAV vector serotypes according to the disclosure are listed below: [000108] - Dystrophinopathies are a spectrum of X-linked muscle diseases caused by pathogenic variants in DMD gene, which encodes the protein dystrophin. Dystrophinopathies comprises Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and DMD-associated dilated cardiomyopathy. [000109] - The Limb-girdle muscular dystrophies (LGMDs) are a group of disorders that are clinically similar to DMD but occur in both sexes as a result of autosomal recessive and autosomal dominant inheritance. Limb-girdle dystrophies are caused by mutation of genes that encode sarcoglycans and other proteins associated with the muscle cell membrane, which interact with dystrophin. The term LGMD1 refers to genetic types showing dominant inheritance (autosomal dominant), whereas LGMD2 refers to types with autosomal recessive inheritance. Pathogenic variants at more than 50 loci have been reported (LGMD1A to LGMD1G; LGMD2A to LGMD2W). Calpainopathy (LGMD2A) is caused by mutation of the gene CAPN3 with more than 450 pathogenic variants described. Contributing genes to LGMD phenotype include: anoctamin 5 (ANO5), blood vessel epicardial substance (BVES),calpain 3 (CAPN3), caveolin 3 (CAV3), CDP-L-ribitol pyrophosphorylase A (CRPPA), dystroglycan 1 (DAG1), desmin (DES), DnaJ heat shock protein family (Hsp40) homolog, subfamily B, member 6 (DNAJB6), dysferlin (DYSF), fukutin related protein (FKRP), fukutin (FKT), GDP-mannose pyrophosphorylase B (GMPPB), heterogeneous nuclear ribonucleoprotein D like (HNRNPDL), LIM zinc finger domain containing 2 (LIMS2), lain A:C (LMNA), myotilin (MYOT), plectin (PLEC), protein O-glucosyltransferase 1 (PLOGLUT1), protein O-linked mannose N-acetylglucosaminyltransferase 1 (beta 1,2-) (POMGNT1), protein O-mannose kinase (POMK), protein O-mannosyltransferase 1 (POMT1), protein O-mannosyltransferase 2 (POMT2), sarcoglycan alpha (SGCA), sarcoglycan beta (SGCB), sarcoglycan delta (SGCD), sarcoglycan gamma (SGCG), titin-cap (TCAP), transportin 3 (TNPO3), torsin 1A interacting protein (TOR1AIP1), trafficking protein particle complex 11 (TRAPPC11), tripartite motif containing 32 (TRIM 32) and titin (TTN). Major contributing genes to LGMD phenotype include CAPN3, DYSF, FKRP and ANO5 (Babi Ramesh Reddy Nallamilli et al., Annals of Clinical and Translational Neurology, 2018, 5, 1574-1587. [000110] - The Emery-Dreifuss Muscular Dystrophy (EDMD) caused by defects in one of the gene including the EMD gene (coding for emerin), the FHL1 gene and the LMNA gene (encoding lamin A and C). [000111] - Nesprin-1 and Nesprin-2 related muscular dystrophy caused by defects in the SYNE1 and SYNE2 gene, respectively; LUMA related muscular dystrophy caused by defects in the TMEM43 gene; LAP1B related muscular dystrophy caused by defects in the TOR1AIP1 gene. [000112] - Facio-scapulo-humeral muscular dystrophy, type 1 (FSHD1A), such as associated with defect in the DUX4 gene (contraction of the D4Z4 macrosatellite repeat in the subtelomeric region of chromosome 4q35) or the FRG1 gene; Facio-scapulo-humeral muscular dystrophy, type 2 (FSHD1B) caused by defects in the SMCHD1 gene. [000113] - Dysferlin is involved in neurological disorders including multiple sclerosis (Hochmeister et al., J. Neuropathol. Exp. Neurol., 2006 Sep;65(9):855-65); Alzheimer (Galvin et al., Acta Neuropathol., 2006 Dec;112(6):665-71 and choreic movement (Takahashi T, et al., Mov. Disord., 2006, Sep;21(9):1513-5).[000114] - Spinal muscular atrophy is a genetic disorder caused by mutations in the Survival Motor Neuron 1 (SMN1) gene which is characterized by weakness and wasting (atrophy) in muscles used for movement. Mutations in ASAH1 gene lead to SMA-PME (spinal muscular atrophy with progressive myoclonic epilepsy). [000115] - Centronuclear myopathies include X-linked myoatubular myopathy (MTM1), autosomal and recessive dominant centronuclear myopathies (DNM2, BIN1, etc) as disclosed in the Tables above. X-linked myotubular myopathy is a genetic disorder caused by mutations in the myotubularin (MTM1) gene which affects muscles used for movement (skeletal muscles) and occurs almost exclusively in males. This condition is characterized by muscle weakness (myopathy) and decreased muscle tone (hypotonia). [000116] - Titinopathies are genetic disorders caused by mutations in the Titin (TTN) gene. Both dominant and recessive TTN mutations have been reported to cause a wide spectrum of cardiac and skeletal muscle diseases. Dominant titinopathies include hereditary myopathy with early respiratory failure (HMERF) caused by mutations in exon 344, and late‐onset tibial muscular dystrophy (TMD). Recessive titinopathies include limb‐girdle muscular dystrophy 2J, young‐ or early‐adult‐onset distal titinopathy, Emery‐Dreifuss‐like myopathy without cardiomyopathy, and congenital myopathy with or without heart disease. [000117] - Pompe disease is a genetic disorder caused by mutations in the acid alpha- glucosidase (GAA) gene. Mutations in the GAA gene prevent acid alpha-glucosidase from breaking down glycogen effectively, which allows this sugar to build up to toxic levels in lysosomes. This buildup damages organs and tissues throughout the body, particularly the muscles, leading to the progressive signs and symptoms of Pompe disease. [000118] - Glycogen storage disease III (GSD3) is an autosomal recessive metabolic disorder caused by homozygous or compound heterozygous mutation in the Amylo-Alpha-1, 6-Glucosidase, 4-Alpha-Glucanotransferase (AGL) gene which encodes the glycogen debrancher enzyme and associated with an accumulation of abnormal glycogen with short outer chains. Clinically, patients with GSD III present in infancy or early childhood with hepatomegaly, hypoglycemia, and growth retardation. Muscle weakness in those with IIIa is minimal in childhood but can become more severe in adults; some patients develop cardiomyopathy.[000119] Glycogen storage disease type IV (GSD IV) is an ultra-rare autosomal recessive disease caused by variants in the GBE1 gene, which encodes the glycogen branching enzyme (GBE). GSD IV accounts for approximately 3% of all GSD. The phenotype of GSD IV ranges from neonatal death to mild adult-onset disease with variable hepatic, muscular, neurologic, dermatologic, and cardiac involvement. [000120] - Genome-wide association studies identified the BIN1 locus as a leading modulator of genetic risk in Alzheimer’s disease (AD) (Voskobiynyk et al., eLife doi: 10.7554 / eLife.57354; July 13, 2020). Hereditary spastic paraplegias (HSPs) are a group of rare, inherited, neurological diseases characterized by broad clinical and genetic heterogeneity. Lower-limb spasticity with first motoneuron involvement is the core symptom of all HSPs. The genes responsible for HSPs include at least 79 SPG genes. Mutations in SPG7 and SPAST are common causes of hereditary spastic paraplegia (HSP) (Review in Lallemant-Dudek P. et al. Fac. Rev., 2021, Mar 10;10:27). [000121] MECP2 (methylCpG binding protein 2) appears to be essential for the normal function of nerve cells. It is an important reader of DNA methylation. Its methyl-CpG-binding (MBD) domain recognizes and binds 5-mC regions. MECP2 gene is X-linked and subject to X inactivation. MECP2 gene mutations are the cause of most cases of Rett syndrome, a progressive neurologic developmental disorder and one of the most common causes of cognitive disability in females. At least 53 disease-causing mutations in this gene have been discovered. [000122] Diseases associated with Androgen Receptor (AR) include Androgen Insensitivity syndrome (partial or complete) and Spinal and bulbar muscular atrophy, X-linked 1 (SMAX1 or SBMA). SBMA is an X-linked, adult-onset neuromuscular condition caused by an abnormal polyglutamine (polyQ) tract expansion in androgen receptor (AR) protein. AR isoform 2 is a naturally occurring variant encoding a truncated AR lacking the polyQ harboring domain. Delivery of this isoform using rAAV9 vector resulted in amelioration of the disease phenotype in SBMA mice by restoring polyQ AR-dysregulated transcription activity (Lim et al., Science Advances, 2021, 7, 34). [000123] In some particular embodiments, the target gene for gene therapy (additive gene therapy or gene editing) is a gene responsible for a neuromuscular disease selected from thegroup comprising : Duchenne muscular dystrophy and Becker muscular dystrophy (DMD gene); Limb-girdle muscular dystrophies (LGMDs) (CAPN3, DYSF, FKRP, ANO5, DNAJB6 genes and others such as SGCA, SGCB, SGCG); Spinal muscular atrophy (SMN1, ASAH1 genes) and Amyotrophic lateral sclerosis (SOD1, ALS2, SETX, FUS, ANG, TARDBP, FIG4, OPTN and others); Myotubular myopathy (MTM1 gene); Centronuclear myopathies (MTM1, DNM2, BIN1 genes); Nemaline myopathies (NEB, ACTA1, KLHL40, KLHL41, KBTBD13, LMOD3, TNNT1, TNNT3, TPM2, TPM3, CFL2, MYPN genes); Selenoprotein N-related myopathy (SEPN1 gene); Congenital myasthenia (ColQ, CHRNE, RAPSN, DOK7, MUSK, CHAT, AGRN genes); Pompe disease (GAA gene); Glycogen storage disease III (GSD3) (AGL gene); Myotonic dystrophy type 1 (DMPK gene) and type 2 (CNBP / ZNF9 gene); Hereditary paraplegia (SPAST (SPG4), SPG7, and other SPG genes such as SPG11, SPG20 and SPG21; in particular SPAST (SPG4) and SPG7 ) Charcot-Marie-Tooth, Type 4B1 (MTMR2); Rett syndrome (MCEP2); and Spinal and bulbar muscular atrophy, X-linked 1 (SMAX1 or SBMA) (AR gene) . [000124] In some preferred embodiments, the target gene is selected from the group consisting of : DMD, CAPN3, DYSF, FKRP, SGCA, SGCB, SGCG, ANO5, MTM1, DNM2, BIN1, GAA, AGL, ColQ, DOK7, SMN1, MCEP2, AR, and ASAH1 genes. [000125] In the context of the invention, a therapeutically effective amount refers to a dose sufficient for reversing, alleviating or inhibiting the progress of the disorder or condition to which such term applies, or reversing, alleviating or inhibiting the progress of one or more symptoms of the disorder or condition to which such term applies. The term "effective dose" or "effective dosage" is defined as an amount sufficient to achieve, or at least partially achieve, the desired effect. [000126] The effective dose is determined and adjusted depending on factors such as the composition used, the route of administration, the physical characteristics of the individual under consideration such as sex, age and weight, concurrent medication, and other factors, that those skilled in the medical arts will recognize. [000127] In the various embodiments of the present invention, the pharmaceutical composition comprises a pharmaceutically acceptable carrier and / or vehicle.[000128] A "pharmaceutically acceptable carrier” refers to a vehicle that does not produce a severe adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to a non-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. [000129] Preferably, the pharmaceutical composition contains vehicles, which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. [000130] The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or suspensions. The solution or suspension may comprise additives which are compatible with viral vectors and do not prevent viral vector particle entry into target cells. In all cases, the form must be sterile and must be fluid to the extent that easy syringe ability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. An example of an appropriate solution is a buffer, such as phosphate buffered saline (PBS) or Ringer lactate. [000131] The invention provides also a method for treating a muscle and / or nervous system disorder, comprising: - administering to a patient in need thereof a therapeutically effective amount of AAV9, AAVrh10 or AAVrh74 vector at an initial time point and then administering to the patient a therapeutically effective amount of AAVpo1 vector at a later time point. [000132] The invention provides also the use of the pharmaceutical compositions according to the present disclosure for the preparation of a medicament for treating a muscle or nervous system disorder, in particular muscle or CNS disorder according to the present disclosure; preferably a muscle and nervous system disorder, in particular muscle and CNS disorder according to the present disclosure.[000133] The invention provides also a product containing an AAVpo1 vector and an AAV9, AAVrh10 or AAVrh74 vector as a combined preparation for sequential use in the treatment of a muscle and / or nervous system disorder. [000134] As used herein, the term “patient” or “individual” includes human and other mammalian subjects that receive either prophylactic or therapeutic treatment. Preferably, a patient or individual according to the invention is a human. [000135] In some embodiments, the individual has been previously tested as having no or low level neutralization antibodies against the AAV serotype used for the initial administration. Neutralizing antibodies against AAV serotype may be measured by standard AAV neutralization assays that are well-known in the art and disclosed in the examples of the present application. In some particular embodiments, low level neutralization antibodies is a titer inferior or equal to 1:10 in the neutralization assay disclosed in the examples of the present application. [000136] Treatment", or "treating" as used herein, is defined as the application or administration of a therapeutic agent or combination of therapeutic agents to a patient, or application or administration of said therapeutic agents to an isolated tissue or cell line from a patient, who has a disease, in particular a muscle disorder with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve or affect the disease, or any symptom of the disease. In particular, the terms "treat' or treatment" refers to reducing or alleviating at least one adverse clinical symptom associated with the disease. [000137] The term "treatment" or "treating" is also used herein in the context of administering the therapeutic agents prophylactically. [000138] The pharmaceutical compositions of the present invention are generally administered according to known procedures, at dosages and for periods of time effective to induce a therapeutic effect in the patient. The pharmaceutical compositions may be administered by any convenient route, such as in a non-limiting manner by infusion or bolus injection, by absorption through epithelial or mucocutaneous linings (e.g., oral mucosa, rectal and intestinal mucosa, etc.).[000139] As used herein, the term “systemic administration” refers to a route of administration of a substance (vector) into the circulatory system and includes enteral or parenteral administration. Parenteral administration includes injection, infusion, implantation and others. [000140] The administration can be systemic, local or systemic combined with local; systemic includes parenteral and oral, and local includes local and loco-regional. Systemic administration is preferably parenteral such as subcutaneous (SC), intramuscular (IM), intravascular such as intravenous (IV), intraarterial or intracoronary; intraperitoneal (IP); intradermal (ID), epidural or else. Local administration is preferably intracerebral, intracerebroventricular, intracisternal, and / or intrathecal administration. The administration may be for example by injection or perfusion. In some preferred embodiments, the administration is parenteral, preferably intravascular such as intravenous (IV), intraarterial or intracoronary, intramuscular, or subcutaneous; more preferably intravascular such as intravenous (IV) or intraarterial, and even more preferably intravenous (IV). In some other preferred embodiments, the administration is intracerebral, intracerebroventricular, intracisternal, and / or intrathecal administration, alone or combined with parenteral administration, preferably intravascular, intramuscular or subcutaneous administration; more preferably intravascular such as intravenous (IV) or intraarterial administration, and even more preferably intravenous (IV). In some other preferred embodiments, the administration is parenteral, preferably intravascular alone or combined with intracerebral, intracerebroventricular, intracisternal, and / or intrathecal administration. The initial time point administration and the later time point administration may be by the same route or a different route. [000141] In some embodiments, the initial time point and the later time point of administration of the combination of vectors are separated by several weeks, months or years. [000142] In some embodiments, the vector used for the initial administration and the vector used for the later time point administration encode the same transgene and the later time point is characterized by a decreased transgene expression level or therapeutic efficacy of the AAV vector administered at the initial time point. According to this embodiment, the sequential administration of the combination of AAV vector serotypes via the systemic route producessustained, high level transgene expression in target organs as compared to the administration of the vector used for the initial administration. [000143] In some other embodiments, the vector used for the initial administration and the vector used for the later time point administration encode different transgenes and the sequential administration of the combination of AAV vector serotypes via the systemic route allows expression of two different transgenes in target organ(s) as compared to the administration of the vector used for the initial administration. [000144] Vector transduction and transgene expression are determined by administration of the combination of AAV vector serotypes, sequentially, via the systemic route in animal models such as mouse models that are well known in the art and disclosed in the examples of the present application. Administration of the vector used for the initial administration, alone is used for comparison. Vector transduction may be determined by measuring vector genome copy number per diploid genome by standard assays that are well known in the art such as real-time PCR assay disclosed in the examples of the present application. Transgene expression is measured at the mRNA or protein levels by standard assays that are well known in the art such as quantitative RT-PCR assay and quantitative western blot analysis as disclosed in the examples of the present application. [000145] The practice of the present invention will employ, unless otherwise indicated, conventional techniques, which are within the skill of the art. Such techniques are explained fully in the literature. [000146] The invention will now be exemplified with the following examples, which are not limitative, with reference to the attached drawings in which: FIGURE LEGENDS [000147] Figure 1: Neutralization antibody titers against rAAVpo1 in mice injected with various rAAV serotypes. A. Mice injected with AAV8, AAV9, AAVrh10, AAVpo1A1, AAV9P1. B. Mice injected with AAvrh74. [000148] Figure 2: Neutralization antibody titers against various rAAV serotypes in mice injected with rAAVpo1[000149] Figure 3: Neutralization antibody titers against rAAVpo1 in rats injected with various rAAV serotypes [000150] Figure 4: Neutralization antibody titers against various rAAV serotypes in rats injected with rAAVpo1 [000151] Figure 5: Seroprevalence of AAV8, AAV9, AAVpo1 (=AAVpo1WT) and AAVpo1.A1 in sera from 50 human donors. A. Seroprevalence in percent. B. Seropositivity in percent. [000152] Figure 6: Biodistribution in skeletal muscles from rats injected with AAV8, AAV9 or AAVrh10 carrying rat Mtm1 gene, followed by AAVpo1.A1-Luciferase gene administration, or rats injected with AAVpo1.A1 carrying rat Mtm1 gene, followed by AAV8-, AAV9- or AAVrh10-Luciferase administration, and controls. A. Vector copy number in Tibialis Anterior. B. Vector copy number in diaphragm. [000153] Figure 7: Luciferase activity in skeletal muscles from rats injected with AAV8, AAV9 or AAVrh10 carrying rat Mtm1 gene, followed by AAVpo1.A1-Luciferase administration, or rats injected with AAVpo1.A1 carrying rat Mtm1 gene, followed by AAV8-, AAV9- or AAVrh10-Luciferase administration, and controls. A. Luciferase activity quantified by Relative Luminescence Unit (RLU) normalized by µg of proteins in Tibialis Anterior. B. Luciferase activity quantified by Relative Luminescence Unit (RLU) normalized by µg of proteins in diaphragm. EXAMPLES EXAMPLE 1 Materials and Methods [000154] The neutralizing antibody assay was performed as previously described (Meliani et al., Hum. Gene Ther. Methods, 2015, 26, 45-53). The 2V6.11 cell line (Accession number CVCL_6355) which inducibly expresses the human adenovirus E4 ORF 634kDa oncoprotein was previously described in Mohammadi et al. (Nucleic Acids Res., 2004, 32, 2652-2659). This cell line was chosen for its highest efficacy of AAV transduction as shown previously in Meliani et al., precited. Recombinant AAV vector serotypes express luciferase reporter geneunder control of CMV promoter. Briefly, on day 1, 96-well plates were seeded with 2 x 1042V6.11 cells for 24 hours in presence of ponasterone A (Life technologies, Carlsbad, USA). A multiplicity of infection (MOI) of 200 was used for each AAV serotype leading to a transduction efficiency of 10,000 to 1000,000 relative light unit (RLU) depending on the serotype. Recombinant AAV-CMV-Luciferase (AAV-CMV-Luc) was diluted in serum-free DMEM (Life technologies, Carlsbad, USA) and incubated with half-logarithmic serial dilutions (1 / 1 to 1 / 3160) of the serum samples, and then incubated for 1 hour at 37°C. Starting point dilution can be adjusted in post-injected samples. Subsequently, the serum-vector mixtures were added to the cells incubated in DMEM with 10% FCS at 37°C and 5% CO2. Each dilution was plated in duplicate. After 24 hours, cells were lysed with the Bright Lite system (Promega, Madison, USA) and the luciferase activity was measured on a luminometer (ENSPIRETM, Perkin Elmer, Waltham, USA). Transduction efficiency was measured as Relative Light Unit (RLU) per second. The neutralizing titer was reported as the highest serum dilution that inhibited AAV transduction by ≥ 50 % compared with the control without serum (100 % transduction). Results 1. Neutralization of rAAVpo1 by sera from mice injected with rAAV from various human or NHP serotypes [000155] AAV vectors (AAVpo1.A1, AAV8, AAV9, AAVrh10) expressing a transgene under the control of Desmin promoter or CAG promoter were injected intravenously at 2 x 1013vg / kg in mice at 3 weeks of age (n= 3-10 per group). AAV9P1 vector was injected intravenously at 5 x 1013vg / kg in 4 mice at 7 weeks of age. AAVpo1.A1 is AAVpo1 wild- type modified with peptide A1, previously disclosed in WO 2021 / 219762. AAVpo1.A1 is representative of AAVpo1. AAV9P1 was previously disclosed in Weinmann et al., Nat Commun. 2020 Oct 28;11(1):5432. doi: 10.1038 / s41467-020-19230-w and WO 2019 / 207132. AAVrh74 vector was injected intravenously at 1012vg / kg or 5x1012vg / kg in mice at 3 weeks of age (n= 4 per group). AAV8 was used for comparison. Sera were collected 6-24 weeks post-injection and neutralizing antibody titers against AAVpo1.A1 and injected serotype were determined.[000156] Mice injected with AAV9, AAVrh10 develop neutralizing antibodies against the injected serotype but do not develop neutralizing antibodies against AAVpo1.A1 (Figure 1A). Similar results were obtained for AAVrh74 (Figure 1B); these results with AAVrh74 were expected since its capsid has 99 % sequence identity with that of AAVrh10. These results indicate that AAVpo1 vector such as AAVpo1.A1 vector could be used for re-administration after an initial administration of AAV9, AAVrh10 or AAVrh74 vector. 2. Neutralization of rAAV from various human or non-human serotypes by sera from mice injected with rAAVpo1 [000157] AAVpo1.A1 vector expressing MTM1 under the control of Desmin promoter were injected intravenously at 2x1013vg / kg in 5 mice at 3 weeks of age. Sera were collected 12 weeks post-injection and neutralizing antibody titers against AAV8, AAV9, AAV5, AAVpo1 wild-type (AAVpo1WT) and AAVpo1.A1 were determined. [000158] Mice injected with AAVpo1.A1 develop high levels of neutralizing antibodies against the injected serotype (AAVpo1.A1) as well as against AAVpo1WT (titer between 1:316 and 1:3160). Neutralizing titers against AAV9 and AAV5 were lower in a range, between 1:3.16 and 1:31.6 (except 1:100 and 1:316 in 2 / 5 mice for AAV9). In contrast, neutralizing titers against AAV8, were much lower (undetectable in 3 / 5 mice; titer of 1:1 in 2 / 5 mice; Figure 2). 3. Neutralization of rAAVpo1 by sera from rats injected with rAAV from various human or NHP serotypes [000159] AAV vectors (AAVpo1.A1, AAV8, AAV9, AAVrh10) expressing a transgene under the control of Desmin promoter were injected intravenously at 2 x 1013vg / kg in rats at 3 weeks of age (n= 5-15 per group). Sera were collected 3-6 weeks post-injection and neutralizing antibody titers against AAVpo1.A1 and injected serotype were determined. [000160] Rats injected with AAV9, AAVrh10 develop neutralizing antibodies against the injected serotype but do not develop neutralizing antibodies against AAVpo1.A1 or at low titer (undetectable in 11-12 / 15 rats; titer of 1:1 in 3-4 / 15 rats; Figure 3). These results indicate that AAVpo1 vector such as AAVpo1.A1 vector could be used for re-administration after an initial administration of AAV9 or AAVrh10 vector.4. Neutralization of rAAV from various human or non-human serotypes by sera from rats injected with rAAVpo1 [000161] AAV vectors (AAVpo1.A1, AAV8, AAV9, AAVrh10) expressing MTM1 under the control of Desmin promoter were injected intravenously at 2x1013vg / kg in 5 rats at 3 weeks of age. Sera were collected 3 weeks post-injection and neutralizing antibody titers against AAV8, AAV9, AAVrh10 and AAVpo1.A1 were determined. [000162] Rats injected with AAVpo1.A1 develop high levels of neutralizing antibodies against the injected serotype (AAVpo1.A1) (titers above 1:1000). Neutralizing titers against AAV9 were lower at a level of 1:100. In contrast, neutralizing titers against AAV8 and AAVrh10, were much lower (for AAV8: titer of 1:1 in 3 / 5 rats, titer of 1:3.16 and 1:10 for 2 / 5 rats; for AAVrh10: undetectable in 2 / 5 rats; titer of 1:1, 1:3,16 and 1:10 in 3 / 5 rats; Figure 4). 5. Neutralization of rAAV from porcine AAV serotype 1 and various human or NHP serotypes by pooled human immunoglobulin [000163] Normal human immunoglobulin (TEGELINE; 50 mg / mL) was tested in neutralizing antibody assay. AAVpo1 (AAVpo1WT and AAVpo1.A1) neutralizing antibody titers were in the range of 1:10 to 1:31.6 while AAV8 and AAV9 neutralizing antibody titers were about 1:316. These results show that neutralizing antibodies against AAVpo1 are present in human sera. 6. AAV seroprevalence in humans [000164] The seroprevalence of AAV8, AAV9 and AAVpo1 (AAVpo1.A1 or AAVpo1WT) was determined on sera from 50 human blood donors randomly selected. AAVpo1 (AAVpo1.A1 and AAVpo1WT) has a lower seroprevalence than AAV8 and AAV9 (Figure 5A). All AAVpo1 (AAVpo1WT or AAVpo1A1) positive donors are AAV8 and AAV9 positive. However, the neutralizing antibody titers against AAVpo1 (AAVpo1WT or AAVpo1A1) are lower than the neutralizing antibody titers against AAV8 and AAV9 (Figure 5B).[000165] Table 1: AAV9 and AAVpo1.A1 neutralizing antibody titers in human donors AAV9 titers Number Number of AAV9 positive sera that are (1 / X) of AAV9 AAVpo1.A1 positive (range = positive sera AAVpo1.A1 titer) 1000 0 0 316 5 4 (1 / 3.16 - 1 / 10) 100 5 4 (1 / 1 - 1 / 3.16) 31,6 7 3 (1 / 1 - 1 / 3.16) 10 10 2 (1 / 1 - 1 / 3.16) 3,16 3 0 1 3 0 TOTAL positives 33 13 Number Number of AAV9 of AAVpo1A1 negative sera negative sera 16 37 [000166] Around 60 % of AAV9 positives patients are AAVpo1 negative (60.4% AAV9 positive / AAVpo1A1 negative (Table 1); 57.6 % AAV9 positive / AAVpo1WT negative) and could therefore be injected with AAVpo1 (AAVpo1WT or AAVpo1A1). In addition, using a positive cut-off inferior or equal to 1:10 in neutralizing antibody titer, 14 / 16 AAV9 positive patients could be injected with AAVpo1.A1 (Table 1). EXAMPLE 2 [000167] AAV vectors expressing a rat Mtm1 cDNA were injected intravenously at a dose of 2x1013vg / kg in 3-week Sprague-Dawley rats and 6 weeks later AAV vectors expressing luciferase were administrated intravenously at a dose of 1x1013vg / kg. The animals were euthanized at 15 weeks of age, 6 weeks after the second injection. Blood samples were collected every 3 weeks and peri-mortem for immunoassays. Muscles and organs were collected and quickly frozen in liquid nitrogen for biochemical assays.Materials and Methods Vector copy number analysis [000168] Samples were homogenized in Cell Lysis Buffer (QIAGEN) by the Bead Mill 24 Homogenizer (FisherBrand) at the speed of 5 m / s for 40 seconds. Total genomic DNA was then extracted with the Gentra Puregen Blood kit (QIAGEN) according to the manufacturer’s instructions. Total gDNA concentration was measured by spectrophotometry using a Nanodrop 8000 (ThermoScientific). The number of vector genomes per diploid genome was quantified from 32 ng of total DNA by Taqman real-time PCR using a LightCycler480 thermocycler (Roche). The titin gene was used for standardization with primers and probe: 5’- gTCCCCTgCgTATCTgCTATg-3’ (forward; SEQ ID NO: 68), 5’- AgAgAggTAgTATTgAAAACgAgCg-3’ (reverse; SEQ ID NO: 69) and 5’- TCCgCAgCTCTAgTggAAgAACCACC-3’ (probe; SEQ ID NO: 70). Primers used for vector genome (Mtm1 transgene) amplification were: 5’- TGGATGGATGGGCGATTTAC-3’ (forward; SEQ ID NO: 71), 5‘-GCGCTGATTGACGAAACTTATC-3’ (reverse; SEQ ID NO: 72) and 5’- TTGAAGAGTACAGAAGGCAGGGCC-3’ (probe; SEQ ID NO: 73) and for Luciferase transgene amplification: 5’-GTGTTGGGCGCGTTATTTATC-3’ (forward; SEQ ID NO: 74), 5’-TAGGCTGCGAAATGTTCATACT-3’ (reverse; SEQ ID NO: 75) and 5’- TTGCGCCCGCGAACGACATTTATA-3’ (probe; SEQ ID NO: 76). Luciferase activity on tissue [000169] Samples were homogenized in PBS and Protease Inhibitor Cocktail (Roche) by the Bead Mill 24 Homogenizer (FisherBrand) at the speed of 4 m / s for 30 seconds. Luciferase activity was measured as Relative Light Unit (RLU) per second on 50 µL lysates on a luminometer (ENSPIRETM, Perkin Elmer, Waltham, USA) after dispensing 100 µL of assay buffer (1mM DTT, 25mM Tris / Base, 1mM EDTA, 15% Glycerol, 8mM MgCl2, 2mM ATP (Roche)) and 100 µL of 167µM D-luciferin (Interchim). The luciferase signal was normalized by the total quantity of proteins determined by BCA protein assay (ThermoFisher Scientific). Results [000170] According to vector genome quantification in the Tibialis Anterior and the diaphragm (Figure 6), AAVpo1A1 vector, administered at a dose of 1x1013vg / kg, effectively transduced skeletal muscles following a first administration of AAV8, AAV9 or AAVrh10vectors at a dose of 2x1013vg / kg, achieving results comparable to a single dose of AAVpo1A1. In contrast, the second administration of AAVpo1.A1 is neutralized by antibodies produced from the first injection of AAVpo1.A1, preventing transduction of the skeletal muscles by the second vector. Similarly, neutralizing antibodies against AAVpo1.A1 detected at 6 weeks after the injection of AAV8 in the serum of one of the rats injected with AAV8 and AAVpo1.A1, led to the absence of transduction of the skeletal muscles by the second administered vector. [000171] The expression of the luciferase gene carried by the second-injected vector, was quantified by measuring the luciferase activity in the Tibialis Anterior and the diaphragm (Figure 7). The luminescence levels were correlated with vector copy numbers: after a first administration of AAV8, AAV9 or AAVrh10 vectors, the injection of AAVpo1A1 allowed the expression of luciferase at levels similar to a single dose of this vector. [000172] These data show that AAVpo1 vectors such as AAVpo1.A1 vector can be used for re-administration following a prior injection of AAV8, AAV9 or AAVrh10 vectors. This also suggests that AAVrh74 can also be used in combination with AAVpo1 vectors as AAVpo1.A1 is not neutralized by antibodies produced after AAVrh74 injection (Figure 1B). [000173] In contrast, injections of AAV8 or AAVrh10 vectors at a dose of 1x1013vg / kg, following a prior injection of AAVpo1.A1, resulted in undetectable levels of skeletal muscle transduction due to the presence of neutralizing antibodies, even at low levels, prior to the second injection (Figure 4 and 6).

Claims

CLAIMS 1. A combination of a recombinant porcine adeno-associated virus serotype 1 (AAVpo1) vector and a recombinant adeno-associated virus serotype 9, rh10 or rh74 (AAV9, AAVrh10, AAVrh74) vector for use in the gene therapy of muscle and / or nervous system diseases in an individual in need thereof, wherein the AAV9, AAVrh10 or AAVrh74 vector is administered at an initial time point and the AAVpo1 vector is administered at a later time point.

2. The combination for use according to claim 1, wherein the AAVpo1, AAV9, AAVrh10 or AAVrh74 serotype is selected from the group consisting of AAV capsids comprising a sequence having at least 95 % identity with any one of SEQ ID NO: 1 and 3 to 5 and hybrid or peptide-modified derivatives thereof having at least 90 % identity with any one of SEQ ID NO: 1 and 3 to 5.

3. The combination for use according to claim 2, wherein the peptide-modified AAV serotype comprises a peptide comprising the sequence RGD, preferably a 7mer peptide comprising the sequence RGD.

4. The combination for use according to claim 2 or claim 3, wherein the peptide- modified AAV serotype comprises a peptide selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82; preferably SEQ ID NO: 6, 15, 33 to 60 and 77 to 82; more preferably SEQ ID NO: 6, 15, 36, 44, 50, 56, 58, 60, 77 and 79.

5. The combination for use according to any one of claims 1 to 4, wherein: - the AAVpo1 serotype has at least 95 % identity with SEQ ID NO: 61 and comprises the peptide A1 of SEQ ID NO: 15; - the AAV9 serotype has at least 95 % identity with SEQ ID NO:3 and comprises a peptide selected from the group consisting of SEQ ID NO: 6 to 60 and 77 to 82, preferably selected from the group consisting of SEQ ID NO: 6, 15, 33 to 60 and 77 to 82 and more preferably SEQ ID NO: 6, 15, 36, 44, 50, 56, 58, 60, 77 and 79; - the AAV9 serotype is hybrid AAV9rh74 having at least 95 % identity with SEQ ID NO: 62; preferably comprising the peptide P1 of SEQ ID NO: 6 or a peptide of any one of SEQ ID NO: 77 to 82; more preferably having at least95 % identity with SEQ ID NO: 64 or 65 and comprising the peptide P1 of SEQ ID NO: 6 or having at least 95 % identity with SEQ ID NO: 83 and comprising the peptide of SEQ ID NO:

77.

6. The combination for use according to any one of claims 1 to 5, wherein the recombinant AAVpo1 vector and the recombinant AAV9, AAVrh74 or AAVrh10 vector encode a transgene of interest for therapy.

7. The combination for use according to claim 6, wherein the transgene encoded by the recombinant AAVpo1 vector and the recombinant AAV9, AAVrh74 or AAVrh10 vector is the same.

8. The combination for use according to claim 6, wherein the transgene encoded by the recombinant AAVpo1 vector and the recombinant AAV9, AAVrh10 or AAVrh74 vector are different.

9. The combination for use according to any one of claims 6 to 8, wherein the transgene of interest for therapy is selected from the group consisting of: (i) therapeutic genes; (ii) genes encoding therapeutic proteins or peptides such as therapeutic antibodies or antibody fragments and genome editing enzymes; and (iii) genes encoding therapeutic RNAs such as interfering RNAs, guide RNAs for genome editing and antisense RNAs capable of exon skipping.

10. The combination for use according to any one of claims 1 to 9, wherein the recombinant AAV serotype for the initial administration and the recombinant AAV serotype for the later administration are chosen respectively from : (i) natural AAV9 capsid of SEQ ID NO: 3 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61 ; (ii) AAVrh10 capsid, in particular natural AAVrh10 capsid of SEQ ID NO: 4 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61 (iii) peptide- modified AAV9 capsid comprising a peptide chosen from SEQ ID NO: 33 to 60 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61; (iv) AAVrh74 capsid, in particular natural AAVrh74 capsid of SEQ ID NO: 5 and AAVpo1 capsid modified with peptide A1 of SEQ ID NO: 61.

11. The combination for use according to any one of claims 1 to 10, which is administered by systemic route, preferably intravascular route, even more preferably intravenous route.

12. The combination for use according to any one of claims 1 to 11, wherein the disease is selected from the group consisting of: neurological diseases, muscular diseases, and combination thereof.

13. The combination for use according to claim 12, wherein the disease is a neuromuscular disease.

14. The combination for use according to any one of claims 1 to 13, wherein the disease is a genetic disease.

15. The combination for use according to claim 14, wherein the target gene for gene therapy of the genetic disease is selected from the group comprising: DMD, DYSF, FKRP, MTM1, SGCA, SGCB, SGCG, CAPN3, ANO5, DNM2, BIN1, GAA, AGL, ColQ, DOK7, SMN1, ASAH1, MCEP2, and AR genes.

16. A product containing a recombinant AAVpo1 vector and a recombinant AAV9, AAVrh10 or AAVrh74 vector as a combined preparation for sequential use in the treatment of a muscle and / or nervous system disorder.

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