Nucleic acid encoding cone-rod homeobox (CRX) protein, and expression vectors thereof, for use in treating a ciliopathy

Abstract

The present invention relates to nucleic acids, expression vectors, and pharmaceutical compositions useful for treating a ciliopathy. The present invention also relates to methods for treating a ciliopathy in a subject in need thereof.

Description

NUCLEIC ACID ENCODING CONE-ROD HOMEOBOX (CRX) PROTEIN, AND EXPRESSION VECTORS THEREOF, FOR USE IN TREATING A CILIOPATHYFIELD OF INVENTION

[0001] The present invention relates to gene therapy.

[0002] The present invention relates to nucleic acids, expression vectors, and pharmaceutical compositions useful for treating a ciliopathy. The present invention also relates to methods for treating a ciliopathy in a subject in need thereof.

[0003] In particular the present invention relates to a gene therapy applicable to the treatment of a ciliopathy; more particularly a CEP290INPHP 1 -related ciliopathy and / or a ciliopathy causing retinal degeneration. The present invention relates especially to a gene therapy applicable to the treatment of retinal ciliopathies, whether non syndromic or syndromic, and more particularly to those associated with CEP290 or NPHP1 mutationsBACKGROUND OF INVENTION

[0004] Ciliopathies are genetic disorders that affect the cellular cilia and / or the cilia anchoring structures, the basal bodies and / or the ciliary function. Such genetic disorders are associated with a wide variety of symptoms, which may affect a number of organs, including (in a non-limitative manner) those associated with visual impairment. This cilium may include the photoreceptor transition zone, known as the photoreceptor connecting cilium, and / or axonemal extension known as the outer segment.

[0005] In fact, the diagnosis of ciliopathies is based on a multidisciplinary approach, which may combine clinical criteria, genetic analyses, and additional investigations. This can include functional assessments of cilia, ophthalmological examinations, and genetic testing.

[0006] Among the ciliary genes involved in ciliopathies, CEP290 is a protein encoded by a gene (i.e., CEP290 in human and Cep290 in mice) that plays a crucial role in the development and function of cilia. In fact, disrupted Cep290 mouse is well known model representative of ciliopathy for skilled person. Mutations in this gene can lead to a wide range of disorders, collectively known as ciliopathies; such as Leber Congenital Amaurosis, Retinitis Pigmentosa, Cone-Rod Dystrophy, Senior-Loken Syndrome, Joubert Syndrome, Nephronophthisis, Meckel-Gruber Syndrome, Bardet-Biedl syndrome. These conditions also affect various organs and systems in the body.

[0007] Such ciliopathies may include, in particular, retinal ciliopathies, such as syndromic retinal disorders and non- syndromic retinal disorders. Retinal ciliopathies represent a subgroup of inherited retinal diseases (IRDs), which can be characterized by structural or functional abnormalities of the photoreceptor cilia.

[0008] In particular, inherited retinal diseases (IRDs) are common causes of visual impairment. They form a group of genetically and clinically heterogeneous diseases. IRDs are characterized by photoreceptor cell death and form a group of clinically and genetically heterogeneous diseases with more than 300 genes identified. For example, IRDs are frequently associated with ciliopathies due to the crucial role of ciliary function in maintaining the integrity of photoreceptor cells. Disruptions in this function can lead to progressive degeneration of the retina and subsequent vision impairment.

[0009] Approximatively 25% of the gene defects underlying retinal degeneration affect the structure or function of the ‘connecting cilium’ in photoreceptors. This structure corresponds to the transition zone of a prototypic cilium, a region with increasing relevance for ciliary homeostasis. The connecting cilium connects the inner and outer segments of the photoreceptor, mediating the bi-directional transport of all proteins required for vision transduction and therefore vision. The outer segment, connecting the cilium and associated basal body, forms a highly specialised sensory cilium dedicated to photoreception and subsequent signal transduction, where the light is converted into a neuro-electrophysiological signal, processed throughout the retina, and transmitted to the brain via the optic nerve.

[0010] Attempts to treat ciliopathies, or to attenuate their progression have been reported in the past. See, for example, Paff et al. (“Current and Future Treatments in Primary Ciliary Dyskinesia”; International Journal of Molecular Sciences; 2021, 22, 9834).

[0011] Reference is further made to WO2020 / 182722 which reports a recombinant AAV vector carrying a nucleic acid sequence encoding the retinal transcription factor cone-rod homeobox (CRX) for use in treating a CRX-associated Inherited retinal diseases (IRD).

[0012] Yet, there remains a need for means for treating ciliopathies, and for treating or preventing and / or reducing the likelihood of occurrence, or the likelihood of reoccurrence, of symptoms thereof.

[0013] In particular, there remains a need for treating ocular ciliopathies, and for treating or preventing and / or reducing the likelihood of occurrence, or the likelihood of reoccurrence of ciliopathies associated with vision impairment; including those associated with photoreceptor impairment or degeneration.

[0014] The invention has for purpose to meet the above-mentioned needs.SUMMARY

[0015] This invention relates to a nucleic acid comprising a sequence encoding a conerod homeobox (CRX) protein or a functional variant thereof, for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof.

[0016] This invention further relates to expression vector for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0017] In some embodiments, the expression vector is a recombinant adeno-associated virus (rAAV) vector.

[0018] In some embodiments, the rAAV vector is a pseudotyped rAAV vector.

[0019] In some embodiments, the rAAV vector comprises an AAV capsid protein from an AAV serotype selected from the group consisting of: AAV2, AAV5 and AAV8.

[0020] This invention further relates to a nanoparticle for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0021] This invention also relates to composition for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; characterized in that it comprises (i) at least one of a nucleic acid as described herein, and / or at least one expression vector as described herein, and / or at least one nanoparticle as described herein, and (ii) a pharmaceutically acceptable excipient.

[0022] In some embodiments, the sequence encoding the CRX protein is operably liked to a promoter allowing the expression in a cone and / or rod photoreceptors.

[0023] In some embodiments, the nucleic acid for use as described herein, the expression vector for use as described herein, the nanoparticle for use as described herein, or the pharmaceutical composition for use as described herein; is administrable via a route selected from the group consisting of: oral, sublingual, buccal, rectal, intravenous, intramuscular, subcutaneous, intranasal, inhalational, vaginal, transdermal, intravitreal, subretinal and suprachoroidal; preferably intravitreal, subretinal and suprachoroidal.

[0024] In some embodiments, the nucleic acid for use as described herein, the expression vector for use as described herein, the nanoparticle for use as described herein, or the pharmaceutical composition for use as described herein; is administrable in a dose ranging from 108copies per dose to 1014copies per dose; more preferably ranging from 109to 3.1012copies per dose.

[0025] In some embodiments, the retinopathy associated to ciliopathy is selected from the group consisting of: Retinal dystrophy (RD) associated to a ciliopathy, Retinitis pigmentosa (RP) associated to a ciliopathy, Leber Congenital Amaurosis (LCA)associated to a ciliopathy, Cone-Rod Dystrophy (CRD) associated to a ciliopathy, Nephronopthisis (NPHP) associated to a ciliopathy, Senior Loken syndrome (SLSN) associated to a ciliopathy, Joubert Syndrome (JBTS) associated to a ciliopathy, Meckel Gruber syndrome (MKS) associated to a ciliopathy, Bardet-Biedl syndrome (BBS) associated to a ciliopathy, Short-rib thoracic dysplasia (SRTD) associated to a ciliopathy, Usher Syndrome associated to a ciliopathy, Alstrdm Syndrome associated to a ciliopathy and Orafaciodigital Syndrome associated to a ciliopathy.

[0026] In some embodiments, the retinopathy associated to a ciliopathy is associated to one or more mutation in one or more gene of Centro somal Protein (CEP), preferably on one or more genes selected from the group consisting of: CEP290, CEP120, CEP104, CEP 55 and. CEP41; and even more preferably CEP290.

[0027] In some embodiments, the retinopathy associated to a ciliopathy is associated to one or more mutation in one or more gene of Nephronophthisis (NPHP), preferably on one or more genes selected from the group consisting of: NPHP I , NPHP2, NPHP3, NPHP4, NPHP5, NPHP6, NPHP7, NPHP8, NPHP9, NPHP10, NPHP11, NPHP12, NPHP 13, NPHP 14, NPHP 15, NPHP 16, NPHP 17, NPHP 18, NPHP 19 and NPHP20; preferably NPHP1 and NPHP6

[0028] In some embodiments, the subject in need thereof is not associated to a mutation in gene CRX.

[0029] In some embodiments, the subject is a mammalian subject, preferably a human subject.

[0030] This invention thus relates to a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use in a method for treating a ciliopathy in a subject in need thereof.

[0031] This invention further relates to an expression vector for use in a method for treating a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0032] In some embodiments, the vector is a recombinant adeno-associated virus (rAAV).

[0033] In some embodiments, the rAAV vector is a pseudotyped AAV.

[0034] In some embodiments, the rAAV vector is characterized in that the AAV capsid is selected from the group consisting of: AAV2, AAV5 and AAV8, preferably an AVV2 / 5 or an AAV2 / 8.

[0035] This invention further relates to a nanoparticle for use in a method for treating a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0036] This invention further relates to a pharmaceutical composition for use in a method for treating a ciliopathy in a subject in need thereof; characterized in that it comprises (i) at least one of a nucleic acid as described hereinabove, and / or at least one expression vector as described hereinabove, and / or at least one nanoparticle as described hereinabove, and (ii) a pharmaceutically acceptable excipient.

[0037] In some embodiments, the sequence encoding the CRX protein is operably liked to a promoter allowing the expression in a cone and / or rod photoreceptors.

[0038] In some embodiments, the nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use as described hereinabove is administrable via a route selected from the group consisting of: oral, sublingual, buccal, rectal, intravenous, intramuscular, subcutaneous, intranasal, inhalational, vaginal, transdermal, intravitreal, subretinal and suprachoroidal; preferably intravitreal, subretinal, and suprachoroidal.

[0039] In some embodiments, the nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use as described hereinabove is administrable in a dose ranging from 108copies per dose to 1014copies per dose; more preferably ranging from 109to 3.1012copies per dose.

[0040] In some embodiments, the ciliopathy is an ocular ciliopathy.

[0041] In some embodiments, the ciliopathy is selected from the group consisting of: hereditary retinopathy, retinal dystrophy, Nephronopthisis (NPHP), Senior-Loken syndrome (SLSN), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Joubert Syndrome (JBTS), and Leber Congenital Amaurosis 10 (LCA-10).

[0042] In some embodiments, the ciliopathy is associated to a mutation in gene CEP290.

[0043] In some embodiments, the subject in need thereof is not associated to a mutation in gene CRX.

[0044] In some embodiments, the subject is a mammalian subject, preferably a human subject.DEFINITIONS

[0045] In the present invention, the following terms have the following meanings:

[0046] “AAV” is an abbreviation for adeno-associated virus, and may be used to refer to the virus itself or derivatives thereof.

[0047] “AAV-CRX refers to an AAV vector suitable for the expression of a CRX protein, or a functional variant thereof.

[0048] “AAV-RK1-CRX” refers to an AAV vector suitable for the expression of a CRX protein, or a functional variant thereof; the nucleic acid sequence suitable for the expression of the CRX protein, or functional variant thereof, being operably linked to a Rhodopsin Kinase 1 promoter, and more preferably to hRKl for human Rhodopsin Kinase 1 promoter. The human rhodopsin kinase (RK) gene promoter is also known as G protein-coupled receptor kinase 1 (GRK1).

[0049] “AAV-GFP refers to an AAV vector suitable for driving the expression of a Green Fluorescent Protein (GFP) protein.

[0050] “AAV-RK1-GFP” refers to an AAV vector suitable for the expression of a GFP protein, or a functional variant thereof; the nucleic acid sequence suitable for the expression of the CRX protein, or functional variant thereof, being operably linked to a Rhodopsin Kinase 1 promoter, and more preferably to hRKl for human Rhodopsin Kinase 1 promoter.

[0051] “Ciliopathies” refers to genetic disorders that affect cellular cilia, including the ciliary axoneme, basal bodies, centrosomes, and transition zone, and / or impair ciliary function. Such genetic disorders are associated with a wide variety of symptoms, which may affect a number of organs, including (in a non-limitative manner) those associated with visual impairment. Within the group of ciliopathies, retinal ciliopathy (also called herein ocular ciliopathy or retinopathy associated to a ciliopathy) refers to retinal disease due to one or more mutation affecting the formation, maintenance and / or functioning of the photoreceptor sensory cilium. This cilium may include the photoreceptor transition zone known as the photoreceptor connecting cilium and axonemal extension known as the outer segment. The diagnosis of ciliopathies is based on a multidisciplinary approach, which may combine clinical criteria, genetic analyses, and additional investigations. This can include functional assessments of cilia, ophthalmological examinations, and genetic testing.

[0052] “CRX”, refers to cone-rod homeobox CRX (NCBI: Nucleotide NM_000554.6; Protein NP_000545.1, or Ensembl No. ENSG00000105392) is an Otd / OTX-like ‘paired’ homeodomain transcription factor that is preferentially expressed in vertebrate rod and cone photoreceptor cells in the retina and pinealocytes in the brain. CRX plays an essential role in the development and maintenance of functional mammalian rod and cone photoreceptors. CRX encodes a 299 amino acid-long protein that contains a homeodomain (HD) near its N-terminus that is responsible for DNA binding. One of skill in the art would understand that a CRX coding sequence may include any nucleic acid sequence that encodes an CRX gene product. The CRX coding sequence may or may not include intervening regulatory elements (e.g.: introns, enhancers, or other non-coding sequences).

[0053] A “CRX functional variant’’ may thus refer to any cone-rod homeobox CRX mutant which retains its function as an Otd / OTX-like ‘paired’ homeodomain transcription factor along with a functional DNA binding domain.

[0054] “Heterologous gene’’ or “heterologous nucleotide sequence’’ will typically refer to a gene or nucleotide sequence that is not naturally occurring in the virus. Alternatively, a heterologous gene or nucleotide sequence may refer to a viral sequence that is placed into a non-naturally occurring environment (e.g.: by association with a promoter with which it is not naturally associated in the virus).

[0055] “Hypomorphic mutation” refers to a mutation that confers a reduced level of activity to the altered gene product or that leads to a reduced level of expression of the altered gene product.

[0056] The term “identity” or “identical”, when used in a relationship between the sequences of two or more amino acid sequences, or of two or more nucleic acid sequences, refers to the degree of sequence relatedness between amino acid sequences or nucleic acid sequences, as determined by the number of matches between strings of two or more amino acid residues or nucleic acid residues. “Identity” measures the percent of identical matches between the smaller of two or more sequences with gap alignments (if any) addressed by a particular mathematical model or computer program (i.e., “algorithms”). Identity of related amino acid sequences or nucleic acid sequences can be readily calculated by known methods. Such methods include, but are not limited to, those described in Lesk A. M. (1988). Computational molecular biology: Sources and methods for sequence analysis. New York, NY: Oxford University Press; Smith D. W. (1993). Biocomputing: Informatics and genome projects. San Diego, CA: Academic Press; Griffin A. M. & Griffin H. G. (1994). Computer analysis of sequence data, Part 1. Totowa, NJ: Humana Press; von Heijne G. (1987). Sequence analysis in molecular biology: treasure trove or trivial pursuit. San Diego, CA: Academic press; Gribskov M. R. & Devereux J. (1991). Sequence analysis primer. New York, NY: Stockton Press; Carillo et al., 1988. SIAM J Appl Math. 48(5): 1073-82. Preferred methods for determining identity are designed to give the largest match between the sequences tested. Methods of determining identity are described in publicly available computer programs. Preferred computer program methods for determining identity between two sequences include theGCG program package, including GAP (Genetics Computer Group, University of Wisconsin, Madison, WI; Devereux et al., 1984. Nucleic Acids Res. 12(1 Pt 1 ):387-95), BLASTP, BLASTN, and FASTA (Altschul et al., 1990. J Mol Biol. 215(3):403-10). The BLASTX program is publicly available from the National Center for Biotechnology Information (NCBI) and other sources (BLAST Manual, Altschul et al. NCB / NLM / NIH Bethesda, Md. 20894). The well-known Smith Waterman algorithm may also be used to determine identity.

[0057] “ITR” or “inverted terminal repeat” refer to the stretch of nucleic acid sequences that exist in AAV vector and / or recombinant Adeno- Associated Viral Vectors (rAAV) that can form a T-shaped palindromic structure, that is required for completing AAV lytic and latent life cycles (Muzyczka N and Berns KI 2001). The term "non- resolvable ITR" refers to a modified ITR such that the resolution by the Rep protein is reduced. A non-resolvable ITR can be an ITR sequence without the terminal resolution site (TRS) which leads to low or no resolution of the non-resolvable ITR and would yield 90-95% of self-complementary AAV vectors (McCarty et al 2003).

[0058] “Operably linked” refers to a functional relationship between two or more polynucleotide (e.g., DNA) segments. Typically, the term refers to the functional relationship of a transcriptional regulatory sequence to a sequence to be transcribed. For example, a promoter or enhancer sequence is operably linked to a coding sequence if it stimulates or modulates the transcription of the coding sequence in an appropriate host cell or other expression system. Generally, promoter transcriptional regulatory sequences that are operably linked to a transcribed sequence are contiguous to the transcribed sequence, i.e., they are cis-acting. However, some transcriptional regulatory sequences, such as enhancers, need not be physically contiguous or located in close proximity to the coding sequences whose transcription they enhance.

[0059] “Pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier” refers to an excipient or carrier that does not produce an adverse, allergic or other untoward reaction when administered to a mammal, such as a human. It includes any and all solvents, such as, for example, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents. A pharmaceuticallyacceptable excipient or carrier refers to a non-toxic solid, semi- solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. For human administration, preparations should meet sterility, pyrogenicity, general safety and purity standards as required by the regulatory offices such as the EMA (European Medicines Agency) or FDA (US Food and Drug Administration).

[0060] “Polynucleotide of interest” refers any nucleotide sequence coding for CRX.

[0061] “Promoter” refers to a sequence that regulates transcription of an operably- linked gene, or nucleotide sequence encoding a protein, etc. Promoters provide the sequence sufficient to direct transcription, as well as the recognition sites for RNA polymerase and other transcription factors required for efficient transcription and can direct cell-specific expression. In addition to the sequence sufficient to direct transcription, a promoter sequence of the invention can also include sequences of other regulatory elements that are involved in modulating transcription (e.g.: enhancers, kozak sequences and introns). In addition, standard techniques are known in the art for creating functional promoters by mixing and matching known regulatory elements. "Truncated promoters" may also be generated from promoter fragments or by mix and matching fragments of known regulatory elements.

[0062] “Subject” or “subject in need thereof’ is refers to a mammal, preferably a human. In one embodiment, a subject may be a “patient”, i.e., a warm-blooded animal, more preferably a human, who / which is awaiting the receipt of, or is receiving medical care or was / is / will be the object of a medical procedure, or is monitored for the development of a disease. In one embodiment, the subject is an adult (for example a subject above the age of 18). In another embodiment, the subject is a child (for example a subject below the age of 18). In one embodiment, the subject is a male. In another embodiment, the subject is a female.

[0063] “Treating” or “treatment” of a given condition, or a clinical symptom thereof, encompasses slowing or arresting or reducing its development or at least one of the clinical symptoms thereof. "Treating" or "treatment" can also refer to alleviating or ameliorating at least one physical parameter including those which may not be discernible by the patient.

[0064] “Preventing” or “prevention” of a given condition, or a clinical symptom thereof, encompasses "preventing or "prevention" as used herein, “’reduction of the likelihood of occurrence” and / or “reduction of a likelihood of re-occurrence”. For example, it may refer to preventing or delaying the onset or development or progression of a given condition. For example, it may refer to reducing the likelihood of occurrence or re-occurrence of the onset or development or progression of a given condition.

[0065] The term "vector" is intended to refer to any polynucleotide construct, typically a plasmid or a viral nucleic acid, used to transmit genetic material to a host cell. Vectors can be, for example, viruses, plasmids, cosmids, or phage. A vector as used herein can be composed of either DNA or RNA. In some embodiments, a vector is composed of DNA.

[0066] An "expression vector" is a vector that is capable of directing, directly or indirectly, the expression of a protein encoded by one or more genes carried by the vector when it is present in the appropriate environment. Vectors are preferably capable of autonomous replication. Typically, an expression vector comprises a transcription promoter, a gene, and a transcription terminator. Gene expression is usually placed under the control of a promoter, and a gene is said to be "operably linked to" the promoter.

[0067] A “recombinant virus vector” or “recombinant viral vector” is intended to refer to a non-wild-type recombinant viral particle that functions as a gene delivery vehicle and which comprises a recombinant viral genome packaged within a viral (e.g.: AAV) capsid. A specific type of virus vector may be a “recombinant adeno-associated virus vector”, or “AAV vector”. The recombinant viral genome packaged in the viral vector is also referred to herein as the “vector genome”.

[0068] As used herein, the term “lipid nanoparticle” or “LNP” refers to a nano-sized transfer vehicle comprising one or more lipids (e.g., cationic lipids, non-cationic lipids, and PEG-modified lipids).

[0069] As used herein, the term “nanoparticle size” or “nanoparticle diameter” refers to the hydrodynamic diameter as measured by dynamic light scattering; for example using a zeta sizer or nanoparticle tracking analysis (NTA).

[0070] As used herein, the term “cationic lipid” refers to any of a number of lipid species that carry a net positive charge at physiological pH. Such lipids include, but are not limited to, DODAC, DOTMA, DDAB, DOTAP, DC-Chol and DMRIE.

[0071] A « lipid aggregate-forming cationic lipid » and the like refer, in the usual and customary sense, to a net positively charged lipid which can facilitate the formation of lipid aggregates. The term « lipid aggregate » refers to a lipid structure including a plurality of lipids or type of lipids, forming a higher order structure (e.g., secondary, tertiary or quaternary structure). Non-limiting examples of lipid aggregates include liposomes, unilamellar vesicles, multilamellar vesicles, micelles, amorphous aggregates, and the like. The lipid aggregates of the present invention can contain any suitable lipid, including cationic lipids, zwitterionic lipids, neutral lipids, or anionic lipids.

[0072] As used herein, the term “ionizable lipid” refers to any of a number of lipid species that are protonated at a low pH (i.e. lower than a physiological pH), which makes them positively charged, but remain neutral at physiological pH.

[0073] As used herein, the term “zwitterionic lipid,” refers to a lipid comprised of a zwitterionic head group and one or more hydrophobic chains linked through one or more covalent bonds.

[0074] As used herein, the term “zwitterionic head group” refers to a multi-functional group, which at a physiological pH is zwitterionic and at other lower pH within the same range is cationic. The cationic and negative charges are either permanent or pH dependent. The cationic nature may arise from any number of primary, secondary, tertiary or quaternary amines, guanadines or non-nitrogenous cations, e.g., phosphonium, sulfonium, etc. The anionic portion can consist of any number of sulfonates, phosphates, phosphonates or carboxylates.

[0075] As used herein, the term “neutral” refers to any of a number of lipid species which exist in an uncharged form. Such lipids include, for example diacylglyceride, tocopherol and cholesterol.

[0076] As used herein, the term “non-cationic lipid” refers to any neutral lipid as described above as well as anionic lipids. Examples of anionic lipids include cardiolipin, diacylphosphatidylglycerol, diacylphosphatidylserine and diacylphosphatidic acid.

[0077] As used herein, the term “encapsulate” refers to coating of various substances within another material at sizes on the nano scale. The encapsulated material is referred to as the internal phase. The encapsulation material is known as the external phase, the shell, coating or membrane. Hence this term may refer to liponanoparticles having an external phase (membrane) including, for example phospholipids, and an internal phase (core).DETAILED DESCRIPTION

[0078] It has now been surprisingly found that expression of the transcription factor CRX can be used for treating a subject carrying a ciliopathy. This is surprising because, to the knowledge of the inventors, the cone-rod homeobox (CRX) protein has never been associated to any form of ciliopathy. Conversely, to the knowledge of the inventors, the occurrence of a mutant form of the CRX gene was never reported in patients diagnosed with a ciliopathy.

[0079] Also surprisingly, it has been found that the expression of CRX in CEP290- deficient mouse models could delay photoreceptor degeneration and promote photoreceptor differentiation. To the knowledge of the inventors, no functional relationship was previously reported between CRX and CEP290, which hence provides evidence of the effect of CRX at a wider level of regulation.

[0080] In particular, the data provided herein enable the treatment of a broad range of ciliopathies, including ciliopathies associated to inherited retinal dystrophies (IRDs), CEP290-associated ciliopathies and beyond CEP290-associated ciliopathies.

[0081] The present invention relates generally to a nucleic acid comprising a sequence to express the CRX protein in subject, expression vectors (such as recombinant viralvectors) and methods to express the CRX protein in subjects suffering from a ciliopathy; in particular of an ocular ciliopathy.

[0082] The present invention relates also to a nucleic acid comprising a sequence to express the CRX protein in subject, expression vectors (such as recombinant viral vectors) and methods to express the CRX protein in subjects suffering from a retinal degeneration in syndromic and non-syndromic ciliopathy.

[0083] According to a first main embodiment, the invention thus relates to a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use in a method for treating a ciliopathy in a subject in need thereof.

[0084] According to a second main embodiment, the invention relates to an expression vector for use in a method for treating a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0085] In some embodiments, a nucleic acid comprising a sequence encoding a conerod homeobox (CRX) protein or a functional variant thereof, for use in a method for treating a retinopathy associated to ciliopathy in a subject in need thereof. Retinopathy associated to ciliopathy is also called herein retinal ciliopathy.

[0086] According to particular embodiments, the expression vector is a recombinant adeno-associated virus (rAAV) vector. According to particular embodiments, the expression vector is a pseudotyped AAV vector.

[0087] The AAV capsid may comprise, consist or substantially consist of an AAV capsid protein selected from a group consisting of: AVI, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV-Rh74, and AAVRhlO, and modified capsids of these serotypes.

[0088] According to particular embodiments, the expression vector is a recombinant adeno-associated virus (rAAV), comprising:(i) an AAV capsid protein, or a mutant form thereof; and(ii) a heterologous polynucleotide encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0089] According to particular embodiments, the expression vector is a recombinant adeno-associated virus (rAAV), comprising:(i) an AAV capsid protein, or a mutant form thereof; and(ii) a heterologous polynucleotide encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, the polynucleotide being operably linked to a promoter suitable for expression in a host cell, and particularly a host mammalian cell.

[0090] According to particular embodiments, the expression vector is a recombinant adeno-associated virus (rAAV); the rAAV being characterized in that the AAV capsid is comprising an AAV capsid protein selected from the group consisting of: AAV2, AAV5 and AAV8, preferably an AAV2 / 5 or an AAV2 / 8. An AAV2 / 5 or AAV2 / 8 vector may thus correspond to an AAV comprising ITR regions corresponding to AAVs from serotype 2, and a capsid from serotype 5 or 8 respectively.

[0091] According to a third main embodiment, the invention relates to a nanoparticle for use in a method for treating a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0092] According to a particular embodiment, the invention relates to a nanoparticle for use in a method for treating a ciliopathy in a subject in need thereof; the nanoparticle consisting essentially of a nucleic acid sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof

[0093] According to a fourth main embodiment, the invention relates to a pharmaceutical composition for use in a method for treating a ciliopathy in a subject in need thereof; characterized in that it comprises(i) at least one of a nucleic acid sequence as defined above, and / or at least one expression vector as defined above, and / or at least one nanoparticle as defined above, and(ii) a pharmaceutically acceptable excipient.

[0094] According to particular main embodiments, the nucleic acid for use, the expression vector for use, the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the nucleic acid sequence encoding the CRX protein is operably liked to a promoter allowing the expression in a cone and / or rod photoreceptors.

[0095] According to particular main embodiments the nucleic acid for use, the expression vector for use, the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that it is administrable via a route selected from the group consisting of: oral, sublingual, buccal, rectal, intravenous, intramuscular, subcutaneous, intranasal, inhalational, vaginal, transdermal, intravitreal, subretinal and suprachoroidal; preferably intravitreal, subretinal, and suprachoroidal.

[0096] According to more particular main embodiments, the invention thus relates to the nucleic acid for use; characterized in that is in a form administrable via a route selected from the group consisting of: intravitreal, subretinal, and suprachoroidal.

[0097] According to more particular main embodiments, the invention thus relates to the expression vector for use; characterized in that is in a form administrable via a route selected from the group consisting of: intravitreal, subretinal, and suprachoroidal.

[0098] According to more particular main embodiments, the invention thus relates to the nanoparticle for use; characterized in that is in a form administrable via a route selected from the group consisting of: intravitreal, subretinal, and suprachoroidal.

[0099] According to more particular main embodiments, the invention thus relates to the pharmaceutical composition for use; characterized in that is in a form administrable via a route selected from the group consisting of: intravitreal, subretinal, and suprachoroidal.

[0100] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that it is administrable in a dose ranging from108copies per dose to 1014copies per dose; more preferably ranging from 109to 5.1012copies per dose.

[0101] According to a particular embodiment, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that it is administrable in a dose ranging from 108, 109, IO10, 1011, 1012, 1013copies per dose to 1014copies per dose.

[0102] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the ciliopathy is selected from the group consisting of: hereditary retinopathy, retinal dystrophy, Nephronopthisis (NPHP), Senior- Loken Syndrome (SLSN), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Joubert Syndrome (JBTS), and Leber Congenital Amaurosis 10 (LCA-10).

[0103] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the ciliopathy is an ocular ciliopathy.

[0104] According to more particular main embodiments, the nucleic acid for use, as defined above, is characterized in that the ciliopathy is an ocular ciliopathy. According to more particular main embodiments, the expression vector for use, as defined above, is characterized in that the ciliopathy is an ocular ciliopathy. According to more particular main embodiments, the nanoparticle for use, as defined above, is characterized in that the ciliopathy is an ocular ciliopathy. According to more particular main embodiments, the pharmaceutical composition for use, as defined above, is characterized in that the ciliopathy is an ocular ciliopathy.

[0105] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the ciliopathy is associated to a mutation in gene CEP290.

[0106] According to another particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the ciliopathy is associated to a mutation in gene NPHP1.

[0107] In some embodiments, the gene CEP290 comprises or consists of the nucleic acid sequence SEQ ID NO:3. In some embodiments, the gene CEP290 consists of the nucleic acid sequence SEQ ID NO:3.

[0108] In some embodiments, the gene NPHP1 comprises or consist to the nucleic acid derived from the mouse gene Nphpl: ENSMUST00000028857.14 from GENCODE VM38 - cDNA RefSeq NM_016902.4.

[0109] According to more particular main embodiments, the nucleic acid for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene CEP290. According to more particular main embodiments, the expression vector for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene CEP290. According to more particular main embodiments, the nanoparticle for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene CEP290. According to more particular main embodiments, the pharmaceutical composition for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene CEP290.

[0110] According to more particular main embodiments, the nucleic acid for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene NPHPE According to more particular main embodiments, the expression vector for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene NPHPE According to more particular main embodiments, the nanoparticle for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene NPHPE According to more particular main embodiments, the pharmaceutical composition for use, as defined above, is characterized in that the ciliopathy is associated to a mutation in gene NPHP1.

[0111] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the subject in need thereof is not associated to (does not bear) a mutation in gene CRX.

[0112] According to more particular main embodiments, the nucleic acid for use, as defined above, is characterized in that the subject in need thereof is not associated to (does not bear) a mutation in gene CRX. According to more particular main embodiments, the expression vector for use, as defined above, is characterized in that the subject in need thereof is not associated to (does not bear) a mutation in gene CRX. According to more particular main embodiments, the nanoparticle for use, as defined above, is characterized in that the subject in need thereof is not associated to (does not bear) a mutation in gene CRX. According to more particular main embodiments, the pharmaceutical composition for use, as defined above, is characterized in that the subject in need thereof is not associated to (does not bear) a mutation in gene CRX.

[0113] According to particular main embodiments, the nucleic acid for use, the expression vector for use, or the nanoparticle for use or the pharmaceutical composition for use as defined above, is characterized in that the subject in need thereof is a mammalian subject, preferably a human subject.

[0114] According to more particular main embodiments, the nucleic acid for use, as defined above, is characterized in that the subject in need thereof is a mammalian subject, preferably a human subject. According to more particular main embodiments, the expression vector for use, as defined above, is characterized in that the subject in need thereof is a mammalian subject, preferably a human subject. According to more particular main embodiments, the nanoparticle for use, as defined above, is characterized in that the subject in need thereof is a mammalian subject, preferably a human subject. According to more particular main embodiments, the pharmaceutical composition for use, as defined above, is characterized in that the subject in need thereof is a mammalian subject, preferably a human subject.

[0115] In some embodiments, the cone -rod homeobox (CRX) protein comprises the amino acid sequence SEQ ID NO:1. In some embodiments, the cone-rod homeobox (CRX) protein consists of the amino acid sequence SEQ ID NO:1.

[0116] In some embodiments, CRX designates a polypeptide belonging to the cone-rod homeobox proteins, by reference to the amino acid sequence of SEQ ID NO: 1, which corresponds to the human CRX amino acid sequence.

[0117] In some embodiments, the CRX polypeptide is the wild-type form of CRX polypeptide.

[0118] In some embodiments, the CRX polypeptide is the wild-type form of human CRX polypeptide.

[0119] In some embodiments, the CRX polypeptide is a variant form of CRX polypeptide.

[0120] In some embodiments, the CRX polypeptide is a variant form of human CRX polypeptide.

[0121] A CRX polypeptide variant may refer to proteins, polypeptides or peptides with the amino acid sequence of which is substantially identical (i.e., largely but not wholly identical) to the sequence of the reference protein, polypeptide, or peptide, e.g., at least about 80% identical or at least about 85% identical, e.g., preferably at least about 90% identical, e.g., at least 91% identical, 92% identical, more preferably at least about 93% identical, e.g., at least 94% identical, even more preferably at least about 95% identical, e.g., at least 96% identical, yet more preferably at least about 97% identical, e.g., at least 98% identical, and most preferably at least 99% identical to the sequence of the protein, polypeptide, or peptide, e.g., to the sequence of a corresponding CRX.

[0122] Accordingly, a protein / polypeptide / peptide having at least about 80% identical to a reference amino acid sequence (e.g. a CRX reference polypeptide of sequence SEQ ID NO: 1) may have at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or even at least 99% of sequence identity with said reference sequence.

[0123] In some embodiments, the CRX polypeptide may be a functional variant thereof, i.e. may be conveniently denoted as "modified", or as "mutated" or "mutant", or as comprising one or more mutations, i.e., comprising one or more amino acid sequences changes compared to the amino acid sequence of CRX that has not been so-mutated, such as, particularly, compared to the amino acid sequence of wild-type CRX. Said variant may comprise the same number of amino acids as any CRX polypeptide defined above, more preferably as SEQ ID NO: 1, and thus the mutations and positions described herein are the same for the variant. Alternatively, said variant may comprise a different number of amino acids as SEQ ID NO: 1. In this case, the skilled artisan in the art will know how to place the mutations and positions described herein in the variant.

[0124] The CRX polypeptide may be a functional variant thereof which comprises or consists of an amino acid sequence of SEQ ID NO: 1 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or even at least 99% of sequence identity with said reference sequence.

[0125] In some embodiments, the CRX polypeptide functional variant may refer to a CRX polypeptide functional variant comprising at least 200 amino-acids, for example at least 210, 220, 230, 240, 250, 260, 270, 280 or 290 amino-acids.

[0126] In particular, the CRX polypeptide functional variant may refer to a CRX polypeptide functional variant comprising at least 150 consecutive amino-acids compared to a reference CRX polypeptide sequence SEQ ID NO 1, in particular at least 200 consecutive amino-acids compared to a reference CRX polypeptide sequence SEQ ID NO 1; for example at least 210, 220, 230, 240, 250, 260, 270, 280 or 290 consecutive amino-acids compared to a reference CRX polypeptide sequence SEQ ID NO 1.

[0127] Accordingly, the CRX polypeptide functional variant may refer to a CRX polypeptide functional variant comprising at least 200 consecutive amino-acids compared to a reference CRX polypeptide sequence SEQ ID NO 1, and which retains its function as an Otd / OTX-like ‘paired’ homeodomain transcription factor along with a functional DNA binding domain.

[0128] According to a main embodiment, the invention relates to a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use for treating a ciliopathy in a subject in need thereof.

[0129] This invention further relates to a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use for treating a ciliopathy in a subject in need thereof, wherein the nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0130] This invention also relates to a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof, wherein the nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0131] In some embodiments, the nucleic acid sequence encoding a cone-rod homeobox (CRX) protein comprises the nucleic acid sequence SEQ ID NO: 2. In some embodiments, the sequence encoding a cone-rod homeobox (CRX) protein consists of the nucleic acid sequence SEQ ID NO:2.

[0132] In some embodiments, the nucleic acid sequence encodes a cone-rod homeobox (CRX) protein susceptible to be encoded by the nucleic acid sequence SEQ ID NO: 2

[0133] This invention also relates to an expression vector for use in a method for treating a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0134] This invention additionally relates to an expression vector for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0135] This invention further relates to an expression vector for use for treating a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, wherein the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0136] This invention supplementary relates to an expression vector for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, wherein the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0137] In some embodiments, the expression vector is a recombinant adeno-associated virus (rAAV) vector.

[0138] In some embodiments, the expression vector is a recombinant adeno-associated (rAAV) vector.

[0139] In some embodiments, the rAAV vector is a pseudotyped rAAV vector.

[0140] In some embodiments, the rAAV vector is characterized in that the AAV capsid is selected from the group consisting of: AAV2, AAV5 and AAV8. In some embodiments, the rAAV vector is selected from the group consisting of: serotype 2, serotype 5 and serotype 8.

[0141] In some embodiments, the rAAV vector comprises the genome of an AAV with a serotype selected from the group consisting of: AAV2, AAV5, and AAV8, preferably AAV2. In this context, the term “genome of the AAV” refers specifically to the Inverted Terminal Repeats (ITRs). This means that rAAV vector is selected from the group consisting of: rAAV serotype 2 inverted terminal repeat, rAAV vector serotype 5 inverted terminal repeat and rAAV serotype 8 inverted terminal repeat.

[0142] In some embodiments, the rAAV vector comprises an AAV capsid protein from an AAV serotype selected from the group consisting of: AAV2, AAV5 and AAV8.

[0143] In some embodiments, the rAAV vector is an AAV2 / 5 or an AAV2 / 8. In fact, the rAAV2 / 5 vector is related to an rAAV vector comprising the genome of AAV2 and the capsid from AAV5. The rAAV2 / 8 vector related to an rAAV vector comprising the genome of AAV2 and the capsid from AAV8.

[0144] This invention further relates to a nanoparticle for use for treating a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0145] This invention additionally relates to a nanoparticle for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0146] This invention also relates to a nanoparticle for use for treating a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, wherein the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0147] This invention relates as well to a nanoparticle for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, wherein the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof allows to restore the ciliary function.

[0148] In some embodiments, the nanoparticle is lipid nanoparticle (LNP).

[0149] In some embodiments, the lipid nanoparticle is selected from the group consisting of: Solid lipid nanoparticles (SLNs), nanostructured lipid carriers (NLCs), liposomes,cationic lipid nanoparticles, PEGylated lipid nanoparticles, ionizable lipid nanoparticles, self-emulsifying drug delivery systems (SEDDS), hybrid lipid-polymer nanoparticles, solid-lipid nanoparticles (SLNs), and cubosomes.

[0150] The lipid nanoparticles (LNP) of the present invention may comprise at least one lipid selected from the group consisting of: a cationic lipid, an ionizable lipid, a noncationic lipid, or conjugate thereof.

[0151] The lipid nanoparticles (LNP) of the present invention may comprise at least one lipid selected from the group consisting of: a cationic lipid, an ionizable lipid, or conjugate thereof.

[0152] According to some embodiments, the lipid nanoparticles (LNP) of the present invention comprise: at least one cationic lipid or ionizable lipid; at least one phospholipid; at least one sterol, for example at least one cholesterol.

[0153] According to some embodiments, the nucleic acid encoding a cone-rod homeobox (CRX) protein or a functional variant thereof is conjugated to the lipid nanoparticle (LNP). According to some embodiments, the said nucleic acid is encapsulated within the lipid nanoparticle (LNP).

[0154] According to some embodiments, the lipid nanoparticles (LNP) of the present invention comprise at least one conjugated lipid; in particular at least one PEGylated lipid.

[0155] According to some embodiments, the lipid nanoparticles (LNP) of the present invention comprise: at least one cationic lipid or ionizable lipid; at least one conjugated lipid, for example at least one PEGylated lipid; at least one phospholipid; at least one sterol, for example at least one cholesterol.

[0156] According to some embodiments, the lipid nanoparticles (LNP) of the present invention comprise:at least one cationic lipid or ionizable lipid; at least one conjugated lipid for example at least one PEGylated lipid; at least one phospholipid; cholesterol.

[0157] According to some embodiments, the lipid nanoparticles (LNP) of the present invention comprise: at least one cationic lipid or ionizable lipid; at least one PEGylated lipid; at least one phospholipid; at least one cholesterol.

[0158] Examples of suitable lipids include, for example, the phosphatidyl compounds (e.g., phosphatidylglycerol, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, sphingolipids, cerebrosides, and gangliosides).

[0159] Also contemplated is the use of polymers as transfer vehicles, whether alone or in combination with other transfer vehicles. Suitable polymers may include, for example, polyacrylates, poly alkylcyanoacrylates, polylactide, polylactide-polyglycolide copolymers, polycaprolactones, dextran, albumin, gelatin, alginate, collagen, chitosan, cyclodextrins, dendrimers and polyethylenimine.

[0160] Examples of cationic lipids include those described in WO 2010 / 053572 or US4897355 or US 5171678 or US5334761 or WO 2010 / 042877 or WO2005 / 121348.

[0161] In some embodiments, the cationic lipid or ionizable lipid is selected from the group consisting of: N-[l-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride (“DOTMA”) dioleoylphosphatidyl-ethanolamine (“DOPE”), 5- carboxyspermylglycinedioctadecylamide (“DOGS”), 2,3-dioleyloxy-N-[2(spermine- carboxamido)ethyl]-N,N-dimethyl-l-propanaminium (“DOSPA”), l,2-Dioleoyl-3- Dimethylammonium- Propane (“DODAP”), 1 ,2-Dioleoyl-3-Trimethylammonium- Propane (“DOT P”), l,2-distearyloxy-N,N-dimethyl-3-aminopropane (“DSDMA”), l,2-dioleyloxy-N,N-dimethyl-3-aminopropane (“DODMA”), l,2-dilinoleyloxy-N,N- dimethyl-3-aminopropane (“DLinDMA”), 1 ,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane (“DLenDMA”), N-dioleyl-N,N-dimethylammonium chloride (“DODAC”), N,N-distearyl-N,N-dimethylammonium bromide (“DDAB”), N-(l,2- dimyristyloxypropyl-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide(“DMRIE”), 3-dimethylamino-2-(cholest-5-en-3-beta-oxybutan-4-oxy)-l-(cis,cis-9,12- octadecadienoxy)propane (“CLinDMA”), 2-[5'-(cholest-5-en-3-beta-oxy)-3'- oxapentoxy)-3-dimethyl l-l-(cis,cis-9',l-2'-octadecadienoxy)propane (“CpLinDMA”), N,N-dimethyl-3,4-dioleyloxybenzylamine (“DMOBA”), l,2-N,N'-dioleylcarbamyl-3- dimethylaminopropane (“DOcarbDAP”), 2,3-Dilinoleoyloxy-N,N- dimethylpropylamine (“DLinDAP”), l,2-N,N-Dilinoleylcarbamyl-3- dimethylaminopropane (“DLincarbDAP”), 1 ,2-Dilinoleoylcarbamyl-3- dimethylaminopropane (“DLinCDAP”), 2,2-dilinoleyl-4-dimethylaminomethyl-[l,3]- dioxolane (“DLin-K-DMA”), 2, 2-dilinoleyl-4-dimethylaminoethyl[ 1,3] -dioxolane( “DLin-K-XTC2-DMA ”), 2-(2,2-di((9Z,12Z)-octadeca-9,12-dien-l-yl)-l,3-dioxolan-4- yl)-N,N-dimethylethanamine ( “DLin-KC2-DMA ”), Nl-[2-((lS)-l-[(3- aminopropyl)amino]-4-[di(3-aminopropyl)amino]butylcarboxamido)ethyl]-3,4- di [oleyloxy] -benzamide (MVL5), N4-cholesteryl-spermine (GL67), (6Z,9Z,28Z,31Z)- heptatriaconta-6,9,28,31- tetraen-19-yl 4-(dimethylamino) butanoate (DLin-MC3- DMA;MC3), di((Z)- non-2- en-1- yl) 9-((4- (dimethylamino)butanoyl)oxy)heptadecanedioate (L319), heptadecan-9- yl 8-((2- hydroxyethyl) (8-(nonyloxy)-8- oxooctyl)amino)octanoate (Lipid 5), heptadecan-9- yl 8- ((2- hydroxyethyl)(6- oxo-6-(undecyloxy)hexyl)amino) octanoate (Lipid H (SM-102)), ((4- hydroxybutyl)azanediyl)bis(hexane-6,l- diyl)bis(2- hexyldecanoate) (ALC-0315), l,l'-((2-(4-(2-((2-(bis(2- hydroxy dodecyl)amino)ethyl) (2- hydroxydodecyl)amino)ethyl)piperazin-l- yl)ethyl)azanediyl) bis(dodecan-2- ol) (C12-200), tetrakis(8- methylnonyl) 3,3',3",3"'-(((methylazanediyl) bis(propane-3,l diyl))bis(azanetriyl))tetrapropionate (3060110), (azanetriyl))tetrapropionate (3060110), 3,6- bis(4-(bis(2- hydroxydodecyl)amino)butyl)piperazine- 2,5- dione (cKK- E12), 3,6- bis(4-(bis((9Z,12Z)-2- hydroxyoctadeca-9,12- dien-l-yl)amino)butyl)piperazine-2,5- dione (OF-02), (((3,6- dioxopiperazine-2,5- diyl)bis (butane-4,1- diyl))bis(azanetriyl))tetrakis(ethane-2,l- diyl) (9Z,9'Z,9"Z,9"'Z,12Z,12'Z,12"Z,12"'Z)- tetrakis (octadeca-9,12-dienoate) (OF-Deg-Lin), (((3,6- dioxopiperazine-2,5- diyl)bis(butane-4,l- diyl))bis(azanetriyl))tetrakis (butane-4,1- diyl) (9Z,9'Z,9"Z,9"'Z,12Z,12'Z,12"Z,12"'Z)- tetrakis (octadeca-9,12- dienoate) (OF-C4-Deg- Lin), N1,N3 ,N5- tris(3-(didodecylamino)propyl)benzene-l,3,5- tricarboxamide (TT3), Hexa(octan- 3- yl) 9,9' ,9" ,9"' ,9"" ,9"'" -((((benzene- 1 ,3,5- tricarbonyl)ris(azanediyl)) tris (propane-3, 1- diyl))tris(azanetriyl))hexanonanoate (FTT5), ethyl 5,5- di((Z)- heptadec-8-en-l-yl)-l-(3- (pyrrolidin-1- yl)propyl)-2,5-dihydro-lH-imidazole-2-carboxylate (A2- Iso5-2DC18); 2- [(polyethylene glycol)-2000]-N,N- ditetradecylacetamide; P- sitosterol, (3S,8S,9S,10R,13R,14S,17R)-17-( (2R,5R)-5- ethyl-6- methylheptan-2- yl)-10,13- dimethyl-2,3,4,7,8,9,10,l l,12,13,14,15,16,17-tetradecahydro-lH- cyclopenta[a]phenanthren-3-ol (ALC-0159); bis(2-(dodecyldisulfa-nyl)ethyl) 3,3'-((3- methyl-9- oxo-10- oxa-13,14- dithia-3,6- diazahexacosyl)azanediyl) dipropionate (BAME-O16B); 2-(((((3S,8S,9S,10R,13R,14S,17R)-10,13- dimethyl- 17-((R)-6- methylheptan-2- yl)-2,3,4,7,8,9,10,l l,12,13,14,15,16,17-tetradecahydro-lH- cyclopenta[a]phenanthren-3-yl)oxy)carbonyl)amino)-N,N-bis(2- hydroxy ethyl)- N- methylethan- 1-aminiumbromide (BHEM-Cholesterol); 1,1 '-((2-(4-(2-((2-(bis(2- hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-l- yl)ethyl)azanediyl)bis(dodecan-2-ol) (C12-200); 3,6-bis(4-(bis(2- hydroxydodecyl)amino)butyl)piperazine-2, 5-dione (cKK-E12); 3P-[N-(N',N'- dimethylaminoethane)- carbamoyl]cholesterol (DC-Cholesterol); 2,3-dioleyloxy- N-[2- (sperminecarboxamido)ethyl]- N,N- dimethyl- 1- propanaminium trifluoroacetate (DOSPA); 1,2- dioleoyl-3- trimethylammonium- propane (DOTAP); 1,2- distearoyl-sn- glycero-3-phosphocholine (DSPC); Ethylphosphatidylcholine (ePC); 1,2-dimyristoyl- rac-glycero-3-methoxypolyethylene glycol-2000 (PEG2000-DMG); Nl,N3,N5-tris(3- (didodecylamino)propyl)benzene-l,3,5-tricarboxamide (TT3); or combinations thereof.

[0162] Examples of ionizable cationic lipid are further described in U.S. provisional patent application 61 / 617,468.

[0163] Also contemplated are cationic lipids such as the dialkylamino-based, imidazole- based, and guanidinium-based lipids. For example, certain embodiments are directed to a composition comprising one or more imidazole-based cationic lipids.

[0164] The use of cholesterol-based cationic lipids is also contemplated by the present invention. Such cholesterol-based cationic lipids can be used, either alone or in combination with other cationic or non-cationic lipids.

[0165] In addition, several reagents are commercially available to enhance transfection efficiency. Suitable examples include LIPOFECTIN (DOTMA:DOPE) (Invitrogen, Carlsbad, Calif.), LIPOFECTAMINE (DOSPA:DOPE) (Invitrogen), LIPOFECTAMINE2000. (Invitrogen), FUGENE, TRANSFECTAM (DOGS), and EFFECTENE.

[0166] In some embodiments, the cationic or ionizable lipids are selected from the group consisting of: distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl- phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N- maleimidomethyl)-cyclohexane-l -carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl- phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1- trans PE, l-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), cholesterol, or a combination thereof.

[0167] According to some embodiments, the lipid nanoparticles (LNP) may comprise phospholipids, sterols, sterol derivatives, and / or polyethylene glycol (PEG) -modified lipids.

[0168] According to some embodiments, the lipid nanoparticles (LNP) may comprise phospholipids selected from the group consisting of: phosphatidylcholine, phosphatidylethanolamine, and derivatives thereof.

[0169] According to some embodiments, the cationic or ionizable lipids may be in combination with one or more phospholipids, sterols, sterol derivatives, and / or polyethylene glycol (PEG)-modified lipids.

[0170] According to some embodiments, compositions comprising lipid nanoparticles of the invention may be supplemented by one or more of the following ingredients: potassium chloride, monobasic potassium phosphate, sodium chloride, dibasic sodium phosphate dihydrate, sucrose tromethamine (tris), tromethamine HC1, acetic acid, sodium acetate.

[0171] Advantageously, the LNP compositions according to the invention may not contain any adenovirus or AAV vector.

[0172] According to some embodiments, lipid nanoparticles according to the invention, and compositions thereof, may be characterized by an average particle size ranging from about 5 nm to about 1000 nm; which may thus rang from about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, to about 1000 nm.

[0173] According to some embodiments, lipid nanoparticles according to the invention, and compositions thereof, may be characterized by an average particle size of less than or equal to about 50nm, less than or equal to about lOOnm, less than or equal to about 250nm, less than or equal to about 500nm, less than or equal to about lOOOnm, or comprising and / or spanning the above numerical ranges.

[0174] In some embodiments, the lipid nanoparticles, and compositions thereof, may be characterized by an average particle size of from about 50nm to 150nm or from about 50nm to about 250 nm.

[0175] In some embodiments, at least 50%, 75%, 80%, 90% (or ranges including and / or spanning the above percentages) of the lipid nanoparticles present have a particle size distribution equal to or less than about: 20nm, 40nm, 60nm, 80nm, lOOnm, l lOnm, 120nm, 130nm, 140nm, 160nm, 180nm, 200nm, 300nm, 400nm, 500nm, or a range including and / or spanning the above values of nm.

[0176] In some embodiments, the lipid nanoparticles, and compositions thereof, have an average particle size of less than or equal to about lOnm, 50nm, lOOnm, 250nm, 500nm, lOOOnm, or comprising and / or spanning the above numerical ranges. In someembodiments, at least 90% of the lipid nanoparticles present have a nanoparticle size distribution equal to or less than about: 20nm, 40nm, 60nm, 80nm, lOOnm, l lOnm, 120nm, 130nm, 140nm, 160nm, 180nm, 200nm, 300nm, 400nm, 500nm, or a range comprising and / or spanning the foregoing values of nm.

[0177] This invention additionally relates to a pharmaceutical composition for use for treating a ciliopathy in a subject in need thereof; characterized in that it comprises (i) at least one of a nucleic acid as defined herein, and / or at least one expression vector as defined herein, and / or at least one nanoparticle as defined herein, and (ii) a pharmaceutically acceptable excipient.

[0178] This invention further relates to a pharmaceutical composition for use for treating a retinopathy associated to a ciliopathy in a subject in need thereof; characterized in that it comprises (i) at least one of a nucleic acid as defined herein, and / or at least one expression vector as defined herein, and / or at least one nanoparticle as defined herein, and (ii) a pharmaceutically acceptable excipient.

[0179] In some embodiments, the pharmaceutical composition further comprises (iii) at least one other active ingredient.

[0180] In some embodiments, the pharmaceutically acceptable excipients that may be used in these compositions include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, proteins (such as, e.g ., serum albumin, gelatin, immunoglobulins and the like), buffer substances (such as, e.g. , phosphates, citrates or other organic acids, and the like), amino acids (such as, e.g. , glycine, glutamine, asparagine, arginine, lysine andthe like), antioxidants (such as, e.g, ascorbic acid and the like), chelating agents (such as, e.g. , EDTA), sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (such as, e.g. , protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate and the like), hydrophilic polymers (such as, e.g. , polyvinylpyrrolidone, poly ethylene-poly oxypropylene block polymers and the like), cellulose-based substances (such as, e.g. ,sodium carboxymethylcellulose), poly acrylates, waxes, nonionic surfactants (such as, e.g. , Tween, pluronics, polyethylene glycol and the like) and wool fat.

[0181] In some embodiments, the pharmaceutical composition according to the present invention comprise vehicles which are pharmaceutically acceptable for a formulation capable of being injected to a subject. 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.

[0182] In some embodiments, the nucleic acid comprising a sequence encoding the CRX protein is operably liked to a promoter allowing the expression in a cone and / or rod photoreceptors.

[0183] In certain embodiments, the promoter is a photoreceptor- specific promoter capable of driving gene expression in rod and / or cone photoreceptors. In some embodiments, the promoter is adaptable to drive expression photoreceptors, preferably in rod and / or cone photoreceptors.

[0184] In some embodiments, the promoter is selected from the group consisting of: Rhodopsin (Rho) promoter, phosphodiesterase (PDE) promoter, retinitis pigmentosa 1 (RP1) promoter, Rhodopsin kinase 1 (GRK1) promoter, rod-specific transcription factor promoter, (Nrl) promoter and cone transducin a-subunit (GNAT2) promoter, preferably GRK1 promoter.

[0185] In some embodiments, the promoter is selected from the group consisting of: promoters that drive expression in rod and / or in cone photoreceptors chosen among Rhodopsin (Rho), phosphodiesterase (PDE), retinitis pigmentosa 1 (RP1), and Rhodopsin kinase 1 (GRK1) promoter.

[0186] In some embodiments, the hGRKl promoter comprises the nucleic acid sequence of SEQ ID NO:4. In some embodiments, the hGRKl promoter consists of the nucleic acid sequence of SEQ ID NO:4.

[0187] In some embodiments, the nucleic acid as described herein, the expression vector as described, or the nanoparticle as described herein, which is administrable via a route selected from the group consisting of: oral, sublingual, buccal, rectal, intravenous, intramuscular, subcutaneous, intranasal, inhalational, vaginal, transdermal, intravitreal, subretinal and suprachoroidal; preferably intravitreal, subretinal, and suprachoroidal.

[0188] In some embodiments, the nucleic acid as described herein, the expression vector as described, or the nanoparticle as described, which is administrable in a dose ranging from 108copies per dose to 1014copies per dose; more preferably ranging from 109to 3.1012copies per dose or from 109to 5.1012copies per dose.

[0189] In some embodiments, the ciliopathy is selected from the group consisting of: Retinal dystrophy (RD), Retinitis pigmentosa (RP), Leber Congenital Amaurosis (LCA, such as LCA10, LCA3, LCA5, LCA6, LCA8 or LCA15), Cone-Rod Dystrophy (CRD). Nephronophthisis (NPHP), Scnior-Lpkcn syndrome (SLSN), Joubert syndrome (JBTS), Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Alstrbm syndrome, Usher syndrome, OrofacioDigital syndrome, Short-rib thoracic dysplasia (SRTD) with or without polydactyly (especially Mainzer-Saldino syndrome and Ellis-van Creveld Syndrome, Jeune Syndrome, Sensenbrenner syndrome), Primary Ciliary Dyskinesia and Polycystic Kidney disease.

[0190] In some embodiments, the ciliopathy is selected from the group consisting of: Retinal dystrophy (RD), Retinitis pigmentosa (RP), Leber Congenital Amaurosis (such as LCA10, LCA3, LCA5, LCA6, LCA8 or LCA15), Nephronophthisis (NPHP), Senior- Lpken syndrome (SLSN), Joubert syndrome (JBTS), Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Alstrbm syndrome, Usher syndrome, OrofaciDigital syndrome, Short-rib thoracic dysplasia (SRTD) with or without polydactyly (especially Mainzer-Saldino syndrome and Ellis-van Creveld Syndrome, Jeune Syndrome and Sensenbrenner syndrome).

[0191] In some embodiments, the ciliopathy is selected from the group consisting of: Nephronophthisis (NPHP), Scnior-Lpkcn syndrome (SLSN), Joubert syndrome (JBTS),Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Alstrom syndrome, Usher syndrome and OrofacioDigital syndrome.

[0192] In some embodiments, the ciliopathy is a ciliopathy affecting the kidney with retinal syndromes, affecting the neurons with retinal syndromes or affecting directly the retina.

[0193] In some embodiments, the ciliopathy is Nephronophthisis-related ciliopathies (NPHP-RC) with retinal syndromes.

[0194] In some embodiments, the ciliopathy is preferably retinal ciliopathy (also called herein ocular ciliopathy or retinopathy associated to a ciliopathy).

[0195] In some embodiments, the ciliopathy is an ocular ciliopathy. In some embodiments, the ciliopathy is a syndromic ocular ciliopathy. In some embodiments, the ciliopathy is a non-syndromic ocular ciliopathy.

[0196] In some embodiments, the ciliopathy is a ciliopathy associated with visual impairment. In some embodiments, the ciliopathy is a ciliopathy not associated with visual impairment.

[0197] In some embodiments, the ciliopathy is selected from the group consisting of: hereditary retinopathy, retinal dystrophy, Leber Congenital Amaurosis (such as LCA10 and LCA3, LCA5, LCA6, LCA8, or LCA15), Nephronopthisis (NPHP), Senior Loken syndrome (SLSN), Joubert Syndrome (JBTS), Meckel Gruber syndrome (MKS), Bardet- Biedl syndrome (BBS), Short-rib thoracic dysplasia (SRTD) with or without polydactyly (especially Mainzer-Saldino syndrome and Ellis-van Creveld Syndrome, Jeune Syndrome, Sensenbrenner syndrome), Usher Syndrome, Alstrom Syndrome, Orafaciodigital Syndrome, Primary Ciliary Dyskinesia, Polycystic Kidney disease.

[0198] In some embodiments, the ciliopathy is selected from the group consisting of: hereditary retinopathy, retinal dystrophy, Leber Congenital Amaurosis (such as LCA10 and LCA3, LCA5, LCA6, LCA8, or LCA15), Senior Loken syndrome (SLSN), Joubert Syndrome (JBTS), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Short-rib thoracic dysplasia (SRTD) with or without polydactyly (especially Mainzer-Saldino syndrome and Ellis-van Creveld Syndrome, Jeune Syndrome, Sensenbrenner syndrome), Usher Syndrome, Alstrdm Syndrome, Orafaciodigital Syndrome, Primary Ciliary Dyskinesia.

[0199] In some particular embodiments, the ciliopathy is selected from the group consisting of: hereditary retinopathy, retinal dystrophy, Nephronopthisis (NPHP), senior loken syndrome (SLSN), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Joubert Syndrome (JBTS), and Leber Congenital Amaurosis 10 (LCA-10).

[0200] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal dystrophy (RD), Retinitis pigmentosa (RP), Leber Congenital Amaurosis (LCA such as LCA10, LCA3, LCA5, LCA6, LCA8 or LCA15), Cone-Rod Dystrophy (CRD), Nephronopthisis (NPHP), Senior Loken syndrome (SLSN), Joubert Syndrome (JBTS), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Short-rib thoracic dysplasia (SRTD) with or without polydactyly (especially Mainzer- Saldino syndrome and Ellis-van Creveld Syndrome, Jeune Syndrome, Sensenbrenner syndrome), Usher Syndrome, Alstrdm Syndrome and Orafaciodigital Syndrome.

[0201] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal dystrophy (RD) associated to a ciliopathy, Retinitis pigmentosa (RP) associated to a ciliopathy, Leber Congenital Amaurosis (LCA) associated to a ciliopathy, Cone-Rod Dystrophy (CRD) associated to a ciliopathy, Nephronopthisis (NPHP) associated to a ciliopathy, Senior Loken syndrome (SLSN) associated to a ciliopathy, Joubert Syndrome (JBTS) associated to a ciliopathy, Meckel Gruber syndrome (MKS) associated to a ciliopathy, Bardet-Biedl syndrome (BBS) associated to a ciliopathy, Short-rib thoracic dysplasia (SRTD) with or without polydactyly associated to a ciliopathy (especially Mainzer-Saldino syndrome and Ellis- van Creveld Syndrome, Jeune Syndrome, Sensenbrenner syndrome), Usher Syndrome associated to a ciliopathy, Alstrdm Syndrome associated to a ciliopathy and Orafaciodigital Syndrome associated to a ciliopathy.

[0202] In some particular embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal dystrophy (RD), Retinitis pigmentosa (RP), senior loken syndrome (SLSN), Meckel Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Joubert Syndrome (JBTS), and Leber Congenital Amaurosis (LCA).

[0203] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal dystrophy (RD), Retinitis pigmentosa (RP), Leber Congenital Amaurosis (LCA), Cone-Rod Dystrophy (CRD), Joubert Syndrome (JBTS), Meckel Gruber syndrome (MKS), Short-rib thoracic dysplasia (SRTD) and Primary Ciliary Dyskinesia.

[0204] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: senior loken syndrome (SLSN), Bardet-Biedl syndrome (BBS), Alstrbm Syndrome, Usher Syndrome and Orafaciodigital Syndrome.

[0205] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal Dystrophy (RD), Retinitis Pigmentosa (RP), Leber Congenital Amaurosis (LCA) (such as LCA10 and LCA3, LCA5, LCA6, LCA8, or LCA 15) and Cone-Rod Dystrophy (CRD).

[0206] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Nephronophthisis (NPHP), Scnior-Lpkcn syndrome (SLSN), Joubert syndrome (JBTS), Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Alstrbm syndrome, Usher syndrome, OrofaciDigital syndrome and other subset of IRDs.

[0207] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Scnior-Lpkcn syndrome (SLSN), Joubert syndrome (JBTS), Leber Congenital Amaurosis (LCA) (such as LCA10 and LCA3, LCA5, LCA6, LCA8, or LCA 15).

[0208] In some embodiments, the retinopathy associated to a ciliopathy is Leber Congenital Amaurosis (LCA), and the LCA is preferably selected from the group consisting of: LCA10, LCA3, LCA5, LCA6, LCA8 and LCA15, more preferably LCA10.

[0209] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Nephronophthisis (NPHP), Scnior-Lpkcn syndrome (SLSN), Joubert syndrome (JBTS), Meckel-Gruber syndrome (MKS), Bardet-Biedl syndrome (BBS), Alstrbm syndrome, Usher syndrome, OrofaciDigital syndrome and subset of IRDs.

[0210] In some embodiments, the retinopathy associated to a ciliopathy is selected from the group consisting of: Leber Congenital Amaurosis, Retinitis Pigmentosa, Cone-Rod Dystrophy, Senior-Loken Syndrome, Joubert Syndrome, Nephronophthisis, Meckel- Griiber Syndrome, Bardet-Biedl syndrome.

[0211] In some embodiments, the ciliopathy is a ciliopathy associated with a mutation in one or more genes selected from the group consisting of: BBS1, BBS2, BBS3 (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), BBS21 (C8ORF37), CEP290, TMEM216, AHI1, NPHP1, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5orf42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP 104, KIAA0556, CELSR2, ARMC9, CEP120, SUFU, PIBF1, B9D1, MKS1, B9D2, TMEM231, MKS1, TMEM216, TMEM67, CEP290, RPGRIP1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231, KIF14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), EXOC3L2, NPHP1 (SLSN type 1), NPHP4 (SLSN type 4), IQCB1 (also known as NPHP5) (SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), SDCCAG8 (SLSN type 7), NPHP1, INVS (also known as NPHP2), NPHP 3, NPHP4, IQCB1 (also known as NPHP5), CEP290 (also known as NPHP6), GLIS2 (also known as NPHP7), RPGRIP1L (also known as NPHP8), NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP 13), ZNF423 (also known as NPHP 14), LCA (such as LCA10,LCA3,LCA5,LCA6,LCASorLCA15(Lebercilin),ABCEl,ABHDll,ABI2,ADGR VI, ALMS1,APLNR,AP4E1,ARL13B,ARL6,B9D1,B9D2, BAMBI, BBS1,BBS2,BBS4,BBS5 ,BBS7,BBS9,BB1P1, CC2D2A, C2ORF71, C8ORF37, CEP104, CEP120, CEP162, CEP164,CEP290, CEP41, CFAP410, CLRN1, C0PS7A, CPIANE1, CSPP1,DCDC2,DYNC2H1,DY NC2I1,DYNC2I2,DYNC2LI1,DYNLT2B,DNASE1L2,EHD1,EVC,EVC2,EDC3,EZR,FA M161A,FGF4,FNDC3B,FOLR1, GCGR, GKN1, GPA33,ICA1,IFT14O,IFT172,IFT27,IFT 43,IFT52,IFT74,IFT80,IFT81,INTU,IQCBl,KATNIP,KIFll,KIF3A,KIF7,KIAA0556,KI AAO586,KIAAO753,KIZ,LCA5,LZTFL1,MAK,MMACHC,MKS1,NACC1,NEK1,NEK2,N PHPl,NPHP2,NPHP3,NPHP4,NPHP5,NPHP6,NPHP7,NPHP8,NPHP9,NPHP10,NP HP11,NPHP12,NPHP13,NPHP14,NPHP15,NPHP16,NPHP17,NPHP18,NPHP19,NP HP20,OFD1,PDE6D,POC1B,PGBD1,PTP4A1,PYG02,RAB28,RP1,RP1L1,RP2,RPGR ,RPGRIP1,RPGRIP1L,SDCCAG8,SRTD1,SRTD12,SH3PXD2A,SIK3,SPATA7,SPRED3 ,SRA1,SUN1, TCTN1, TCTN2, TCTN3, TOPORS, TMEM107, TMEM138, TMEM160, TME M216, TMEM231, TMEM237, TTC21B, TTC8, TULP1, WDR19, WDR34, WDR35.

[0212] In some embodiments, the ciliopathy is a ciliopathy associated with a mutation in one or more genes selected from the group consisting of: BBS1, BBS2, BBS3 (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), BBS21 (C8ORF37), CEP290, TMEM216, AHI1, NPHP1, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5orf42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP 104, KIAA0556, CELSR2, ARMC9, CEP120, SUFU, PIBF1, B9D1, MKS1, B9D2, TMEM231, MKS1, TMEM216, TMEM67, CEP290, RPGRIP1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231, KIF14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), EXOC3L2, NPHP1 (SLSN type 1), NPHP4 (SLSN type 4), IQCB1 (also known as NPHP5) (SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), SDCCAG8 (SLSN type 7), NPHP1, INVS (also known as NPHP2), NPHP3, NPHP4, IQCB1 (also known as NPHP5), CEP290 (also known as NPHP6), GLIS2 (also known as NPHP7), RPGRIP1L (also known as NPHP8), NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP13), and ZNF423 (also known as NPHP14).

[0213] In some embodiments, the ciliopathy or the retinopathy associated to a ciliopathy is a ciliopathy associated with a mutation in one or more genes selected from the group consisting of: BBS1, BBS2, BBSS (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBSS 3 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), BBS21 (C8ORF37), CEP290, TMEM216, AHI1, NPHP1, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5orf42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP104, KIAA0556, CELSR2, ARMC9, CEP120, SUFU, PIBF1, B9D1, MKS1, B9D2, TMEM231, MKS1, TMEM216, TMEM67, CEP290, RPGRIP1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231, KIF14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), EXOC3L2, NPHP1 (SLSN type 1), NPHP4 (SLSN type 4), IQCB1 (also known as NPHP5) (SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), SDCCAG8 (SLSN type 7), NPHP1, INVS (also known as NPHP2), NPHP3, NPHP4, IQCB1 (also known as NPHP5), CEP290 (also known as NPHP6), GLIS2 (also known as NPHP7), RPGRIP1L (also known as NPHP8), NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP13), ZNF423 (also known as NPHP14), LCA (such as LCA10,LCA3,LCA5,LCA6,LCASorLCA15(Lebercilin),ABCEl,ABHDll,ABI2,ADGR VI,. ALMS1,APLNR,AP4E1,ARL13B,ARL6,B9D1,B9D2, BAMBI, BBS1,BBS2,BBS4,BBS5 ,BBS7,BBS9,BBIPl,CC2D2A,C2ORF71,C8ORF37,CEP104,CEP120,CEP162,CEP164, CEP290, CEP41, CFAP410, CLRN1, COPS7A, CPLANE1, CSPP1,DCDC2,DYNC2H1,DY NC2I1,DYNC2I2,DYNC2LI1,DYNLT2B,DNASE1L2,EHD1,EVC,EVC2,EDC3,EZR,FA M161A,FGF4,FNDC3B,FOLR1, GCGR, GKN1, GPA33,ICA1,IFT14O,IFT172,IFT27,IFT 43,IFT52,IFT74,IFT80,IFT81,INTU,IQCBl,KATNIP,KIFll,KIF3A,KIF7,KIAA0556,KI AA0586, KIAAO753,KIZ,LCA5,LZTFL1, MAK, MMACHC,MKS1,NACC1,NEK1,NEK2,N PHPl,NPHP2,NPHP3,NPHP4,NPHP5,NPHP6,NPHP7,NPHP8,NPHP9,NPHP10,NPHP11,NPHP12,NPHP13,NPHP14,NPHP15,NPHP16,NPHP17,NPHP18,NPHP19,NP HP20,OFD1,PDE6D,POC1B,PGBD1,PTP4A1,PYG02,RAB28,RP1,RP1L1,RP2,RPGR ,RPGRIP1,RPGRIP1L,SDCCAG8,SRTD1,SRTD12,SH3PXD2A,SIK3,SPATA7,SPRED3,SRA1,SUN1, TCTN1, TCTN2, TCTN3, TOPORS, TMEM107, TMEM138, TMEM160, TMEM216, TMEM231, TMEM237, TTC21B, TTC8, TULP1, WDR19, WDR34, WDR35.

[0214] In some embodiments, the ciliopathy or the retinopathy associated to a ciliopathy is a ciliopathy associated with one or more mutations in one or more genes of NPHP (Nephronophthisis), preferably on one or more genes selected from the group consisting of: NPHP I. NPHP2, NPHP3, NPHP4, NPHP 5, NPHP6, NPHP7, NPHP8, NPHP9, NPHP 10, NPHP11, NPHP 12, NPHP13, NPHP 14, NPHP15, NPHP 16, NPHP 17, NPHP 18, NPHP19 and NPHP20; preferably NPHP1 and NPHP6.

[0215] In some embodiments, the ciliopathy or the retinopathy associated to a ciliopathy is a ciliopathy associated with one or more mutations in one or more genes of CEP (Centrosomal Protein), preferably on one or more genes selected from the group consisting of: CEP290, CEP162, CEP164, CEP120, CEP104, CEP55 and CEP41 preferably CEP290, CEP162, CEP164, CEP120, CEP104, and CEP41; and even more preferably CEP290.

[0216] In some embodiments, the Bardet-Biedl syndrome (BBS) is associated with a mutation in one or more genes selected from the group consisting of: BBS1, BBS2, BBS3 (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), and BBS21 (C8ORF37).

[0217] In some embodiments, the Joubert syndrome (JBTS) is associated with a mutation in one or more genes selected from the group consisting of: CEP290, TMEM216, AHI1, NPHP1, TMEM67, RPGR1P1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5orf42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP104, KIAA0556, CELSR2, ARMC9, CEP120, SUFU, P1BF1, B9D1, MKS1, B9D2, and TMEM231.

[0218] In some embodiments, the Meckel-Gruber syndrome (MKS) is associated with a mutation in one or more genes selected from the group consisting of: MKS1, TMEM216, TMEM67, CEP290, RPGR1P1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231,K1F14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), and EXOC3L2.

[0219] In some embodiments, the Senior-L0ken syndrome (SLSN) is associated with a mutation in one or more genes selected form the group consisting of: NPHP1 (SLS type 7), NPHP4 (SLS type 4), IQCB1 (also known as NPHP5) SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), and SDCCAG8 (SLS type 7).

[0220] In some embodiments, the Nephronopthisis (NPHP) is associated with a mutation in one or more genes selected from the group consisting of: NPHP1, INVS (also known as NPHP2), NPHP3, NPHP4, IQCB1 (also known as NPHP5). CEP290 (also known as NPHP6). GLIS2 (also known as NPHP7). RPGRIP1L (also known as NPHP8). NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP 10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP13), ZNF423 (also known as NPHP14), NPHP15, NPHP16, NPHP17, NPHP18, NPHP19 and NPHP20. In said embodiment, the Nephronopthisis (NPHP) may be associated with retinal syndromes.

[0221] In some embodiments, the ciliopathy is associated to a mutation in gene CEP290. In some embodiments, the ciliopathy is not associated to a mutation in gene CEP290.

[0222] In some embodiments, the retinopathy associated to a ciliopathy is associated to a mutation in gene CEP290. In another embodiment, the retinopathy associated to a ciliopathy is not associated to (does not bear) a mutation in gene CEP290.

[0223] Advantageously, the ciliopathy is not associated to (does not bear) a mutation in a gene selected from the group consisting of: PDE6B, NMNAT1, ARL13b, AIPL1 and ABCA4 genes.

[0224] In some embodiments, the ciliopathy is not associated to a mutation in gene CRX.

[0225] In some embodiments, the retinopathy associated to a ciliopathy is not related to (does not bear) a mutation in CRX gene.

[0226] In one embodiment, the mutation is a homozygote mutation. In a second embodiment, the mutation is a heterozygote mutation.

[0227] In one embodiment, the ciliopathy is a recessive genetic disorder. Thus, two copies of the mutated gene are required to cause the ciliopathy.

[0228] In a second embodiment, the ciliopathy is a dominant genetic disorder. Thus, only one copy of the mutated gene is required to cause the ciliopathy.

[0229] In some embodiments, the subject is a mammalian subject.

[0230] In one embodiment, the subject is non-human mammalian subject. In another embodiment, the subject is a human.

[0231] The subject, preferably the mammalian subject, and more preferably the human, may comprise a subject at any developmental stage, including a neonate (newborn), infant, toddler, child, adolescent, or adult (which encompass young adult and elderly individual).

[0232] In some embodiments, the mammalian subject is preferably a toddler, child, adolescent, or adult, more preferably an adolescent or an adult, even more preferably an adult.

[0233] In some embodiment, the subject is selected from the group consisting of: Mice, rats, guinea pigs, rabbits, hamsters, monkeys, pigs, dogs, cats, ferrets, sheep, primates.

[0234] In some embodiments, the subject in need thereof is a subject having one or more mutation selected from the group consisting of: BBS1, BBS2, BBS3 (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), BBS21 (C8ORF37), CEP290, TMEM216, AHU, NPHP1, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5orf42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP104, KIAA0556, CELSR2, ARMC9, CEP120, SUFU, PIBF1, B9D1, MKS1, B9D2, TMEM231, MKS1, TMEM216, TMEM67,CEP290, RPGRIP1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231, K1E14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), EXOC3L2, NPHP1 (SLS type 1), NPHP4 (SLS type 4), IQCB1 (also known as NPHP5) (SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), SDCCAG8 (SLS type 7), NPHP1, INVS (also known as NPHP2), NPHP3, NPHP4, IQCB1 (also known as NPHP5), CEP290 (also known as NPHP6), GLIS2 (also known as NPHP7), RPGRIP1L (also known as NPHP8), NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP13), ZNF423 (also known as NPHP14) LCA (such as LCA10,LCA3,LCA5,LCA6,LCASorLCA15(Lebercilin),ABCEl,ABHDll,ABI2,ADGR VI, ALMS1,APLNR,AP4E1,ARL13B,ARL6,B9D1,B9D2, BAMBI, BBS1,BBS2,BBS4,BBS5 ,BBS7,BBS9,BBIPl,CC2D2A,C2ORF71,C8ORF37,CEP104,CEP120,CEP162,CEP164, CEP290, CEP41, CFAP410, CLRN1, COPS7A, CPLANE1, CSPP1,DCDC2,DYNC2H1,DY NC2I1,DYNC2I2,DYNC2LI1,DYNLT2B,DNASE1L2,EHD1,EVC,EVC2,EDC3,EZR,FA M161A,FGF4,FNDC3B,FOLR1, GCGR, GKN1, GPA33,ICA1,IFT14O,IFT172,IFT27,IFT 43,IFT52,IFT74,IFT80,IFT81,INTU,IQCBl,KATNIP,KIFll,KIF3A,KIF7,KIAA0556,KIAAO586,KIAAO753,KIZ,LCA5,LZTFL1,MAK,MMACHC,MKS1,NACC1,NEK1,NEK2,N PHPl,NPHP2,NPHP3,NPHP4,NPHP5,NPHP6,NPHP7,NPHP8,NPHP9,NPHP10,NP HP11,NPHP12,NPHP13,NPHP14,NPHP15,NPHP16,NPHP17,NPHP18,NPHP19,NP HP20,OFD1,PDE6D,POC1B,PGBD1,PTP4A1,PYG02,RAB28,RP1,RP1L1,RP2,RPGR ,RPGRIP1,RPGRIP1L,SDCCAG8,SRTD1,SRTD12,SH3PXD2A,SIK3,SPATA7,SPRED3 ,SRA1,SUN1, TCTN1, TCTN2, TCTN3, TOPORS, TMEM107, TMEM138, TMEM160, TME M216, TMEM231, TMEM237, TTC21B, TTC8, TULP1, WDR19, WDR34, WDR35.

[0235] In some embodiments, the subject in need thereof is a subject having one or more mutation selected from the group consisting of: BBS1, BBS2, BBS3 (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (IFT27), BBS20 (IFT72), BBS21 (C8ORF37), CEP290, TMEM216, AHI1, NPHP1, TMEM67, RPGRIP1L, ARL13B, CC2D2A, OFD1, INPP5E, TMEM138, C5or 42, TCTN1, TCTN2, TCTN3, ZNF423, CEP41, TMEM237, CSPP1, PDE6D, IFT172, KIAA0586, TXNDC15, CEP104, KIAA0556, CELSR2, ARMC9,CEP120, SUFU, P1BF1, B9D1, MKS1, B9D2, TMEM231, MKS1, TMEM216, TMEM67, CEP290, RPGR1P1L, CC2D2A, NPHP3, TCTN2, B9D1, B9D2, TMEM231, KIF14, TMEM107, TXNDC15, C5orf42, CSPP1, CEP55, SEC8 (also known as EXOC4), EXOC3L2, NPHP1 (SLS type 1), NPHP4 (SLS type 4), IQCB1 (also known as NPHP5) (SLS type 5), CEP290 (also known as NPHP6) (SLS type 6), SDCCAG8 (SLS type 7), NPHP1, INVS (also known as NPHP2), NPHP 3, NPHP4, IQCB1 (also known as NPHP5), CEP290 (also known as NPHP6), GLIS2 (also known as NPHP7), RPGRIP1L (also known as NPHP8), NEK8 (also known as NPHP9), SDCCAG8 (also known as NPHP10), TMEM67 (also known as NPHP11), TTC21B (also known as NPHP12), WDR19 (also known as NPHP13), ZNF423 (also known as NPHP14).

[0236] In some embodiments, the subject in need thereof is a subject having one or more mutations one or more gene selected from the group consisting of: CEP, NPHP and BBS.

[0237] In some embodiments, the subject in need thereof is a subject having one or more mutations in the gene CEP (Centrosomal Protein), preferably on one or more gene selected from the group consisting of: CEP290, CEP162, CEP164, CEP120, CEP104, CEP55 and CEP41; preferably CEP29C). CEP 162. CEP 164. CEP120, CEP104, and CEP41; and even more preferably CEP290.

[0238] In some embodiments, the subject in need thereof is a subject having one or more mutations in the gene NPHP, preferably on one or more gene selected from the group consisting of: NPHP1, NPHP2, NPHP3, NPHP4, NPHP 5, NPHP6, NPHP7, NPHP8, NPHP9, NPHP10, NPHP11, NPHP12, NPHP13, NPHP14, NPHP15, NPHP16, NPHP 17, NPHP18, NPHP 19 and NPHP20; preferably NPHP1 and NPHP6.

[0239] In some embodiments, the subject in need thereof is a subject having one or more mutations in the gene BBS, preferably on one or more gene selected from the group consisting of: BBS1, BBS2, BBSS (ARL6), BBS4, BBS5, BBS6 (MKKS), BBS7, BBS8 (TTC8), BBS9, BBS10, BBS11 (TRIM32), BBS12, BBS13 (MKS1), BBS14 (CEP290), BBS15 (WDPCP), BBS16 (SDCCAG8), BBS17 (LZTFL1), BBS18 (BBIP1), BBS19 (1FT27), BBS20 (IFT72), and BBS21 (C8ORF37).

[0240] In some embodiments, the subject in need thereof is a subject having one or more mutations in the gene CEP290 and / or NPHP1.

[0241] In one embodiment, the subject in need thereof is a subject having one or more mutation in the gene CEP290, preferably Cep290~ / ~, more preferably cep290del35 / del35or C(fp2. ()Q^e6 / del36

[0242] In one embodiment, the subject in need thereof is a subject having one or more mutations in the gene NPHP1, preferably NphpE3' .

[0243] According to a main embodiment, the invention relates to compositions comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, expression vectors comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, nanoparticles comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, and pharmaceutical compositions comprising (i) at least one of nucleic acid as described herein, and / or at least one expression vector as described herein, and / or at least one nanoparticle as described herein, and (ii) a pharmaceutically acceptable excipient ; for use in in a method for treating a ciliopathy, in a subject in need thereof.

[0244] In some embodiments, the invention relates to compositions comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, expression vectors comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, nanoparticles comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, and pharmaceutical compositions comprising (i) at least one of nucleic acid as described herein, and / or at least one expression vector as described herein, and / or at least one nanoparticle as described herein, and (ii) a pharmaceutically acceptable excipient ; for use for treating a retinopathy associated to a ciliopathy, in a subject in need thereof.

[0245] The invention also relates to a method of treatment of a ciliopathy as described herein, in a subject in need thereof comprising a step of administering the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein.

[0246] The invention also relates to a method of treatment of a retinopathy associated to a ciliopathy as described herein, in a subject in need thereof comprising a step of administering the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein.

[0247] The invention also relates to a method of treatment of a ciliopathy associated to retinal degeneration as described herein, in a subject in need thereof comprising a step of administering, to said subject, the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein.

[0248] The invention also relates to a method of treatment of a CEP290-associated ciliopathy as described herein, in a subject in need thereof comprising a step of administering, to said subject, the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein.

[0249] The invention also relates to a method of treatment of a NPHP1 -associated ciliopathy as described herein, in a subject in need thereof comprising a step of administering, to said subject, the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein.

[0250] The invention also relates to a use of the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein, for the manufacture of a medicament for treating ciliopathy as described herein.

[0251] The invention also relates to a use of the nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein, for the manufacture of a medicament for treating a retinopathy associated to a ciliopathy as described herein.

[0252] Thus, the invention also relates to a use of nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein, for the manufacture of a medicament to treat a ciliopathy associated to retinal degeneration as described herein.

[0253] Thus, the invention also relates to a use of nucleic acid as described herein, the expression vector as described herein, the nanoparticle as described herein, or the pharmaceutical composition as described herein, for the manufacture of a medicament to treat a CEP290-associated ciliopathy as described herein, in a subject in need thereof.

[0254] Hence, the invention relates to the administration of a subject in need thereof of a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, expression vectors comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, nanoparticles comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, and pharmaceutical compositions, as described herein, wherein the subject has a ciliopathy.

[0255] The invention also relates to the administration of a subject in need thereof of a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, expression vectors comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, nanoparticles comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, as described herein, and pharmaceutical compositions, as described herein, wherein the subject has a retinopathy associated to a ciliopathy.

[0256] This invention further relates to a method of treating a ciliopathy in a subject in need thereof comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0257] This invention also relates to a method to restore ciliary function in a subject in need thereof comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

[0258] This invention additionally relates to a use of a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof for treating ciliopathy in a subject in need thereof.

[0259] Furthermore, this invention relates to the use of a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof for treating ciliopathy in a subject in need thereof, wherein the ciliary function is restored.

[0260] In some embodiments, the nucleic acid may be contained within an expression vector, which may, in turn, be incorporated into a nanoparticle.

[0261] In some embodiments, the nucleic acid, the expression vector, or the nanoparticle may be incorporated into a pharmaceutical composition.BRIEF DESCRIPTION OF THE DRAWINGS

[0262] Figure 1 is a immunohistochemical analysis. VAR002 was injected or not in P14 Cep290flel35 / del35(also named herein: Cep290del / delmice). 1A. Staining of the full retinas and IB. retinal sections from samples collected at P45. All samples were immunolabelled with anti-Rhodopsin (Rho) and anti-cone arrestin antibodies (CA). Nuclei are stained with DAPI. Specific cone staining was performed, in the central block, the retinal sections were immunolabelled with anti m-opsin antibodies (m-opsin, red). Nuclei are stained with DAPI. in the right block, the retinal sections were immunolabelled with anti s-opsin antibodies (s-opsin). Nuclei are stained with DAPI. ONL, outer nuclear layer, INL: inner nuclear layer, GCL: ganglion cell layer. NT: non treated. Bar represents 25pm.

[0263] Figure 2 is an immunohistochemical analysis of the full retinas. VAR002 was subretinally injected or not in P14 Cep290del35 / del35mice. All samples were immunolabelled with anti-CRX (CRX), anti-Rhodopsin (Rho) and anti-cone arrestin antibodies (CA). Nuclei are stained with DAPI. Bar 200 um.

[0264] Figure 3 is an immunohistochemical analysis. AAV-CRX (VAR002) or AAV- GFP was injected in P14 Cep290del35 / de35mice. Retinal sections from samples collected at P45. All samples were immunolabelled with anti-CRX (CRX), anti-Rhodopsin (Rho) to label rod photoreceptors. Cone photoreceptors were labeled with anti-cone arrestin (CA), anti M-opsin (Opnlmw, red), anti S-opsin antibodies (Opnlsw). Anti-Cep290 antibody was used to see the effect on the Cep290 hypomorph mutant (Cep290). Nuclei are stained with DAPI. ONL, outer nuclear layer, INL: inner nuclear layer. Bar represents 20pm.

[0265] Figure 4 is an immunohistochemical analysis. Subretinal injection of AAV-CRX (VAR002) at P14 in Cep290del33 / (M33mice (exon 35 in mice corresponds to exon 36 in humans, as exon 1 in humans is non-coding). As control, the contralateral eye was not injected. Retinal sections from samples collected at P45. All samples were immunolabelled with anti-CRX (CRX), rods were labelled with anti-Rhodopsin (Rho) and cones with anti-cone arrestin (CA), with anti-M-opsin (Opnlmw), anti-S-opsin antibodies (Opnlsw). Nuclei are stained with DAPI. ONL, outer nuclear layer, INL: inner nuclear layer. Bar represents 15pm.

[0266] Figure 5 is an immunohistochemical analysis. Subretinal injection of AAV-CRX (VAR002) at P9 in Nphpl'3' mice and Nphpl+ / ' mice. As control, the contralateral eye was not injected. Retinal sections from samples collected at P28. All samples were immunolabelled with anti-Rhodopsin (Rho) and anti-cone arrestin (CA), to label respectively rods and cones. Nuclei are stained with DAPI. ONL, outer nuclear layer, INL: inner nuclear layer, GCL: ganglion cell layer. Scale Bar: 20pm.EXAMPLES

[0267] The present invention is further illustrated by the following examples. Unless stated otherwise, the acronym “VAR002” will refer herein to an AAV vector defined as AAV2 / 5.hGRKl.CRX; hence referring to an AAV2 / 5 vector suitable for expressing a functional CRX polypeptide, the expression of the functional CRX polypeptide being operably linked to a human GRK1 promoter.

[0268] Example 1: AAV-CRX protects Cep290del / delphotoreceptors degeneration.Materials and MethodsIn-vivo sub-retinal AAV injections

[0269] Adult / young mice were anesthetized with intraperitoneal injection, using 50 pL / 10 g per mice of a solution composed of xylazine (1 mg / mL) and ketamine (10 mg / mL) in phosphate-buffered saline (PBS). Iris were dilated using specific eye drops (Mydriaticum (Tropicamide 5 mg 0.5%) and Neosynephrine (Phenylephrine hydrochloride 2.5%)). Eyes were anesthetizing using Cebesine 0.4%.

[0270] Once anesthetized, Cep290del35 / del35mice were injected into the subretinal space with AAV2 / 5.hGRKl.CRX (VAR002) at post natal day 14 (P14) with IxlO10vg per eye (n=8 mice).

[0271] We evaluate this in a mouse model, Cep290flel35 / del35, which carries a homozygous in-frame Cep290 mutation that skips exon 35 in the mouse, corresponding to exon 36 in the human ortholog. This mutation alters the formation of the photoreceptor lightsensitive primary cilium, leading to rudimentary outer segments (OS) and associated progressive photoreceptor loss, as demonstrated by the gradual reduction in the thickness of the photoreceptor nuclear layer (outer nuclear layer, ONL), with little or no change observed in other cellular layers. Notably, in the Cep290del33 / (M33mouse model, retinal degeneration begins with eye opening, around postnatal day 15 (Pl 5), resulting in a reduction of ONL thickness by half by P30, which is complete by P120. Consistent with the rudimentary OS, scotopic and photopic (electroretinogram) ERG responses are significantly lower than in control mice, alongside a decreased content ofphototransduction proteins such as rhodopsin and cone opsins as early as P15. By P120, most photoreceptors are lost, resulting in flat scotopic and photopic ERG recordings. This mouse model is especially representative of retinal ciliopathy.Histological analysis

[0272] Mice were euthanized using cervical dislocation. Eyes were collected and fixed in 4% paraformaldehyde (PFA) for 1 hour, rinsed in PBS before dehydratation and paraffin inclusion. 7 pm-thick sections were cut with a microtome (Rotatory microtom, ref.: HM 340E [ThermoFisher Scientific™]), placed overnight at 37°C, and then stored at room temperature. Prior to immunohistochemistry (IHC), slides were deparaffinized and then incubated in hot citrate buffer (pH 6, 0.1 M) for 20 minutes at 500 W. Once cooled down, slides were placed in PBS. For immunostaining (IHC) on flat mount retina, the retina was dissected and fixed in 4% PFA for 1 hour, rinsed in PBS and incubated directly with the primary antibody.

[0273] Primary antibodies in Dako REAL™ Antibody diluent (ref S2022 lot 20060091) supplemented with 0.3% Triton X-100 were added to the slides overnight at 4°C in a dry chamber (mouse anti-rhodopsin clone 4D2, ref.: MABN15 [Merck™], dilution 1 / 1000; rabbit anti-cone arrestin, ref.: AB 15282 [Merck™], dilution 1 / 1000; rabbit anti-s opnlsw, ref.: ab5407 [Abeam™], dilution 1 / 1000, rabbit anti-s opnlmw, ref.: ab5405 [Abeam™], dilution 1 / 1000.

[0274] They were rinsed 3 times with PBST, 5 minutes before adding secondary antibodies (donkey anti-mouse IgG Alexa Fluor 555, ref.: A31570 [ThermoFisher Scientific], dilution 1 / 1000; donkey anti-rabbit IgG Alexa Fluor 488, ref.: A21206 [ThermoFisher Scientific™], dilution 1 / 1000; donkey anti-goat Alexa Fluor 488, ref.: Al 1055 [ThermoFisher Scientific™], dilution 1 / 1000) in Dako REAE (tm) Antibody diluent (ref S2022 lot 20060091) supplemented with 0.3% Triton X-100. After 2 hours of incubation at room temperature in the dark, slides were rinsed 3 times with PBST for 5 minutes, and DAPI (ref.: 62248 [ThermoFisher Scientific™], dilution 1 / 1000) was added for 20 minutes in PBST. Finally, slides were rinsed 3 times with PBST. Flat mount retinaswere incubated for 1 night with the secondary antibody.

[0275] FluorSaveTM Reagent (ref.: 345789, Millipore, batch 3034632) was used to add a coverslip. Slides were stored at 4°C. Pictures were acquired using an Imager M2 microscope (Zeiss™) or a Axiozoom Apotome2 (Zeiss™) and analyzed using Zen software and Image J.Results

[0276] To evaluate the potential therapeutic effects of VAR002, subretinal injections were administered at P14 (IxlO10vg), followed by retinal harvesting at post natal day 45 (P45).

[0277] In this disease, photoreceptors do not differentiate properly (lack of opsin expression) including a lack of OS formation. This is characterized by the lack of OS formation. As a consequence, or in addition photoreceptors degenerate. Death of photoreceptor leads to a thinning of the outer nuclear layer (ONL), where the photoreceptors are located. Following subretinal injection, Immunohistochemical analysis did show a significant improvement in the outer nuclear layer (ONL) thickness in treated eyes, and it revealed a notable increase in the expression of photoreceptor markers, including rhodopsin, cone-arrestin, s-opsin and m-opsin (Figure 1A and B).

[0278] Mosaic imaging of the whole retina section of treated Cep2 0del33 / (M33mice showed a large area of transduced photoreceptors expressing hCRX (Figure 2). In this area, CA and Rho expression were observed. This is in contrast with the non-injected retina, where no CRX was observed, and Rho and CA were poorly expressed. This was confirmed with higher magnification (Figure 3).

[0279] In addition, the formation of the outer segment of the photoreceptors was observed after treatment (Figure 3-4). These findings provide evidence that VAR002 promotes photoreceptor maturation by enhancing the expression of key proteins involved in phototransduction and growth of the outer segment. Notably, a more stable expression and localized expression of CEP290 protein on the apical side of the ONL (Figure 4).Furthermore, subsequent to the treatment, the formation of photoreceptor outer segments was observed, with evidence of an increase in well-differentiated cones and rods in the treated retinas (Figures 1-4). Accordingly, VAR002 is effective in promoting photoreceptor maturation, at least in part, through the promotion of expression of key proteins essential for phototransduction and outer segment developmentv

[0280] Altogether these results reflect the correction of fundamental ciliary defects, including the restoration of the transition zone architecture, normalization of intraflagellar transport, and enhancement of cellular proteostasis, which shows an amelioration on the ciliary function.Example: AAV-CRX promotes rod and cone photoreceptor maturation and preservation in Nphpl~'~ retinas

[0281] Materials and Methods

[0282] In-vivo sub-retinal AAV injections

[0283] Nphpl^ and Nphpl+ / ~ 9-10 days old pups (Post-natal days 9-10 P9-P10) were anesthetized with intraperitoneal injection, using 50 pL / 10 g per mice of a solution composed of xylazine (1 mg / mL) and ketamine (10 mg / mL) in phosphate-buffered saline (PBS). For each the right eyelid was opened, and the iris were dilated using specific eye drops (Mydriaticum (Tropicamide 5 mg 0.5%) and Neosynephrine (Phenylephrine hydrochloride 2.5%)). Eyes were anesthetized using Cebesine 0.4%.

[0284] Once anesthetized, Nphpl^' and Nphpl+ / ' mice were injected into the subretinal space with AAV2 / 5.hGRKl.CRX (VAR002) at P14 with 0.5xl010vg per eye (n=10 mice).

[0285] We assessed the therapeutic potential of AAV-CRX (VAR002) in an additional ciliopathy model, the Nphpl^' mouse, which lacks NPHP1 expression. In contrast to CEP290, which is positioned more distally along the connecting cilium toward the outer segment, NPHP1 is part of the NPHP complex located at the base of the connecting ciliumnear the transition zone. It maintains ciliary organization and photoreceptor polarity with the outer segment. Inactivation of NPHP1 leads to structural defects in primary cilia, including destabilization of the axonemal bundle and ciliary junction. These defects impair the ability of cilia to maintain structural integrity. As a consequence, photoreceptors do not grow outer segments and Nphpl^' mice display rapid retinal degeneration with early onset between PIO and P14. By P21, most photoreceptors degenerate, with only two to three rows of photoreceptors remaining. During photoreceptor development and maturation, no outer segments develop due to impaired cilia formation. Little or no change was observed in other cellular layers. Consistent with such photoreceptor development and degeneration, scotopic and photopic (electroretinogram) ERG responses are significantly lower than in control mice. No phenotype is observed in Nphpl+ / ' mouse retina, which corresponds to normal phenotype.

[0286] Histological analysis

[0287] Mice were euthanized using cervical dislocation. Eyes were collected and fixed in 4% paraformaldehyde (PFA) for 1 hour, rinsed in PBS before dehydration and OCT inclusion. 12 pm-thick sections were cut with a cryostat, dried and stored at -80°C. Prior to IHC, slides were defrost and rinsed three time in PBS. For IHC, primary antibodies in PBS with 0.3% Triton X-100 were added to the slides overnight at 4°C in a dry chamber (mouse anti-rhodopsin clone 4D2, ref.: MABN15 [Merck™], dilution 1 / 1000; rabbit anticone arrestin, ref.: AB15282 [Merck™], dilution 1 / 1000; rabbit anti-s opnlsw, ref.: ab5407 [Abeam™], dilution 1 / 1000, rabbit anti-s opnlmw, ref.: ab5405 [Abeam™], dilution 1 / 1000.

[0288] They were rinsed 3 times with PBST, 5 minutes before adding secondary antibodies (donkey anti-mouse IgG Alexa Fluor 555, ref.: A31570 [ThermoFisher Scientific], dilution 1 / 1000; donkey anti-rabbit IgG Alexa Fluor 488, ref.: A21206 [ThermoFisher Scientific™], dilution 1 / 1000; donkey anti-goat Alexa Fluor 488, ref.: Al 1055 [ThermoFisher Scientific™], dilution 1 / 1000) in Dako REAE (tm) Antibody diluent (ref S2022 lot 20060091) supplemented with 0.3% Triton X-100. After 2 hours of incubation at room temperature in the dark, slides were rinsed 3 times with PBST for 5 minutes, and DAPI (ref.: 62248 [ThermoFisher Scientific™], dilution 1 / 1000) was addedfor 20 minutes in PBST. Finally, slides were rinsed 3 times with PBST. Flat mount retinas were incubated for 1 night with the secondary antibody.

[0289] FluorSaveTM Reagent (ref.: 345789, Millipore, batch 3034632) was used to add a coverslip. Slides were stored at 4°C. Pictures were acquired using an Imager M2 microscope (Zeiss™) or a Axiozoom Apotome2 (Zeiss™) and analyzed using Zen software and Image J.

[0290] Results

[0291] To evaluate the potential therapeutic effects of VAR002, subretinal injections were administered at P9-P10 (0.5xl010vg) in Nphpl^' and Nphpl+ / ' mice and the retinas were analyzed at P30. C57B16 / J mouse retina was used as positive control for antibody validation. Nphpl^' photoreceptors do not differentiate properly with a lack of OS formation and therefore degenerate, leading to a thinning of the outer nuclear layer (ONL) where the photoreceptors are located. In contrast, heterozygous mice display normal retinas. Nphpl^' subretinal injection of AAV-CRX led to some preservation of the ONL in contrast with the fellow non-injected eye. Notably, immunohistochemical analysis revealed a notable increase in the expression of rod and cone photoreceptor markers, including rhodopsin, cone-arrestin, compared to the contralateral non-treated eye (Figure 5). In addition, the formation of the outer segment of the photoreceptors was observed after treatment. These findings provide evidence of VAR002 efficacy on Nphpl^' mutant mice. It delays photoreceptor degeneration and enhances photoreceptor maturation by promoting outer segment growth and restoring light-driven translocation of key phototransduction and structural proteins from the inner segment to the outer segment, demonstrating functional intraflagellar transport, which is linked to an amelioration on the ciliary function.SEQUENCES

Claims

CLAIMS1. A nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof, for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof.

2. An expression vector for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the expression vector comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

3. The expression vector for use according to claim 2, wherein the expression vector is a recombinant adeno-associated virus (rAAV) vector.

4. The expression vector for use according to claim 3, wherein the rAAV vector is a pseudotyped rAAV vector.

5. The expression vector for use according to claim 3 or claim 4, wherein the rAAV vector comprises an AAV capsid protein from an AAV serotype selected from the group consisting of: AAV2, AAV5 and AAV8.

6. A nanoparticle for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; the nanoparticle comprising a nucleic acid comprising a sequence encoding a cone-rod homeobox (CRX) protein or a functional variant thereof.

7. A pharmaceutical composition for use in a method for treating a retinopathy associated to a ciliopathy in a subject in need thereof; characterized in that it comprises (i) at least one of a nucleic acid according to claim 1, and / or at least one expression vector according to any of claims 2 to 5, and / or at least one nanoparticle according to claim 6, and (ii) a pharmaceutically acceptable excipient.

8. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 7; whereinthe sequence encoding the CRX protein is operably liked to a promoter allowing the expression in a cone and / or rod photoreceptors.

9. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 8; which is administrable via a route selected from the group consisting of: oral, sublingual, buccal, rectal, intravenous, intramuscular, subcutaneous, intranasal, inhalational, vaginal, transdermal, intravitreal, subretinal and suprachoroidal; preferably intravitreal, subretinal and suprachoroidal.

10. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 9; which is administrable in a dose ranging from 108copies per dose to 1014copies per dose; more preferably ranging from 109to 3.1012copies per dose.

11. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 10; wherein the retinopathy associated to a ciliopathy is selected from the group consisting of: Retinal dystrophy (RD) associated to a ciliopathy, Retinitis pigmentosa (RP) associated to a ciliopathy, Leber Congenital Amaurosis (LCA) associated to a ciliopathy, Cone-Rod Dystrophy (CRD) associated to a ciliopathy, Nephronopthisis (NPHP) associated to a ciliopathy, Senior Loken syndrome (SLSN) associated to a ciliopathy, Joubert Syndrome (JBTS) associated to a ciliopathy, Meckel Gruber syndrome (MKS) associated to a ciliopathy, Bardet- Biedl syndrome (BBS) associated to a ciliopathy, Short-rib thoracic dysplasia (SRTD) associated to a ciliopathy, Usher Syndrome associated to a ciliopathy, Alstrdm Syndrome associated to a ciliopathy and Orafaciodigital Syndrome associated to a ciliopathy.

12. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 11; wherein the retinopathy associated to a ciliopathy is associated to one or more mutation in one or more gene of Centrosomal Protein (CEP), preferably on one ormore genes selected from the group consisting of: CEP290, CEP 164, CEP 162, CEP120, CEP104, CEP55 and. CEP41; and even more preferably CEP290.

13. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 11; wherein the retinopathy associated to a ciliopathy is associated to one or more mutation in one or more gene of Nephronophthisis (NPHP), preferably on one or more genes selected from the group consisting of: NPHP1, NPHP2, NPHP3, NPHP4, NPHP 5, NPHP6, NPHP7, NPHP8, NPHP9, NPHP 10, NPHP11, NPHP12, NPHP13, NPHP14, NPHP 15, NPHP16, NPHP 17, NPHP 18, NPHP19 and NPHP20; preferably NPHP1 and NPHP6.

14. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 13; wherein the subject in need thereof is not associated to a mutation in gene CRX.

15. The nucleic acid for use, the expression vector for use, the nanoparticle for use, or the pharmaceutical composition for use according to any one claims 1 to 14; wherein the subject is a mammalian subject, preferably a human subject.

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