Use of CRB1 gene in treatment of systemic diseases such as diseases in eyes and intestinal tract

Abstract

Provided is use of CRB1 gene in preparing a medicament for treating systemic diseases such as diseases in the eyes and the intestinal tract. Further provided is use of a CRB1 gene-based therapeutic drug in preparing a medicament for treating systemic diseases such as diseases in the eyes and the intestinal tract.

Description

Application of CRB1 gene in the treatment of systemic diseases such as eye and intestinal diseases Technical Field The present invention belongs to the field of biomedicine, and specifically relates to the use of the CRB1 gene in treating systemic diseases such as eye and intestinal diseases. Background Art The Crb1 gene, first discovered in Drosophila, is crucial for maintaining normal apical and basal cell polarity and the integrity of the embryonic epithelium. The phenotype of retinopathy caused by Crb1 mutations varies greatly depending on their temporal and spatial distribution, genetic background, and external factors such as light exposure, with no clear genotype-phenotype correlation documented. Additional genetic and / or environmental factors may influence the expression of CRB1-related diseases. Whether Crb1 plays a similar role in regulating cell adhesion and polarization in mammals remains unknown. The CRB complex has four core components: Crb, Stardust (Sdt), PATJ (Protein associated with tight junctions or Pals1-associated tightjunction protein), and Lin-7. Crb is a transmembrane protein with a large extracellular structure consisting of 29 epidermal growth factor-like (EGF-like) repeats and 4 laminin-A globular structure-like (Lamiin AG-like) repeats. The intracellular structure is smaller and consists of 37 amino acids.

[0027] Sdt is composed of two Lin-2 / Lin-7 (L27) domains, a PDZ domain, a Src-homology 3 (SH3) domain, and a guanylate kinase (GUK) domain. PATJ is composed of four PDZ domains and an N-terminal L27 domain. Lin-7 is a relatively short protein consisting of 195 amino acids, containing an N-terminal L27 domain and a C-terminal PDZ domain. The human body has multiple mucosal epithelia that form a barrier between the external and internal host environments. The gastrointestinal mucosa is a semipermeable barrier with a complex task, allowing nutrient absorption and immune sensing while limiting the transport of potentially harmful antigens and microorganisms. Regulation of this seemingly contradictory task is achieved through interactions between intestinal mucosal structural components and molecules that operate in a dynamic manner to maintain intestinal integrity and immune homeostasis. The function of the intestinal barrier can be compromised by structural damage to the mucosa or subtle changes in barrier regulatory components. The intestinal mucosa is composed of multiple elements, primarily an outer mucus layer (containing commensal intestinal microbiota, antimicrobial proteins, and secretory immunoglobulin molecules); a central monolayer of specialized epithelial cells; and an inner lamina propria (containing innate and adaptive immune cells, such as T cells, B cells, macrophages, and dendritic cells). The mucus layer is the first physical line of defense encountered by foreign molecules upon reaching the intestinal lumen, preventing bacteria from directly contacting epithelial cells. The mucus layer is primarily composed of highly glycosylated mucins, which form a gel-like sieve-like structure covering the intestinal epithelium. In both the small and large intestines, mucin 2 is the most abundant mucin secreted by goblet cells. Mucin 2 knockout mice spontaneously develop colitis, demonstrating that mucin 2 expression is crucial for disease prevention. Beneath the mucus layer lies the intestinal epithelial cell layer, the strongest physical barrier. A pool of multipotent stem cells located in the crypts gives rise to five distinct cell types: absorptive enterocytes, goblet cells, enteroendocrine cells, Paneth cells, and microfold cells. Together, these cells form a continuous, polarized monolayer that separates the lumen from the lamina propria. Because the cell membrane is impermeable to hydrophilic solutes without the presence of specific transport proteins, the passage of such molecules through the intestinal epithelium is highly restricted. The uptake of lipophilic or large molecules primarily depends on diffusion and endocytosis. Molecular transport between intestinal epithelial cells is regulated by the presence of junctional complexes. The three most important junctional complexes are tight junctions (TJs), adherens junctions (AJs), and desmosomes. Tight junctions are the apical junctional complexes of epithelial cells and primarily function to seal intercellular spaces. Tight junctions are composed of transmembrane proteins (such as claudins and occludin), peripheral membrane proteins (such as ZO-1 and ZO-2), and regulatory proteins. Adherens junctions lie beneath tight junctions and, together with desmosomes, provide strong adhesive forces to maintain epithelial structural integrity. Both tight and adherens junctions are connected by linker rings of actin and myosin, which allow for the regulation of junctions by the cytoskeleton. Summary of the Invention In one aspect, the present application provides the use of the CRB1 gene in preparing a drug for treating a disease. In one aspect, the present application provides use of a CRB1 gene therapy drug in the preparation of a drug for treating a disease. Through some technical solutions of the present application, intestinal diseases and / or eye diseases can be treated in a targeted and localized manner, which is more effective than existing methods and has less impact on other organs. Through some technical solutions of the present application, in addition to intestinal diseases, other diseases of the patient, such as eye diseases, can also be treated simultaneously. BRIEF DESCRIPTION OF THE DRAWINGS FIG1 is a diagram of the AAV2 vector used to express mouse Crb1. Figure 2: Retinal phenotypes in Rd8-GF and Rd8-GF-SPF mice. (A, B) Representative immunofluorescence staining images (A) and quantitative analysis (B) of IBA1+ (red) microglia / macrophages in the ONL of WT-SPF, Rd8-SPF, and Rd8-GF mice retinas. n = 3 retinas per group; Student's t-test, ***P < 0.001; NS: not significant. (C) Representative fundus images of Rd8-SPF and Rd8-GF mice at the indicated ages. (D) Histological images of the retina of Rd8-GF mice at the indicated ages. (E) Representative immunofluorescence staining images of ZO-1 (red) and phalloidin (green) in the retinas of 4-week-old mice. (F) Histological images of retinal lesions at P15 and 8 weeks in Rd8-GF mice housed in an SPF environment (referred to as Rd8-GF-SPF mice). SPF: specific pathogen-free; GF: germ-free. DETAILED DESCRIPTION In order to further illustrate the technical means and effects adopted by this application to achieve the intended purpose, the specific implementation methods, structures, features and effects of this application are described in detail below in combination with the accompanying drawings and preferred embodiments. Terms and Definitions To facilitate understanding of the features and benefits of the present invention by those skilled in the art, the following provides a general description and definition of terms and expressions used in the specification and claims. Unless otherwise indicated, all technical and scientific terms used herein have the ordinary meanings as understood by those skilled in the art regarding the present invention. In the event of conflict, the definitions in this specification shall prevail. As used herein, singular terms refer to one or more than one. For example, "element" or "an element" refers to one element or more than one element. As used herein, the term "plurality" refers to at least two. As used herein, the terms "comprise," "include," "have," "contain," or any similar terms are open-ended conjunctions intended to encompass non-exclusive inclusion, indicating that a combination (e.g., a device, composition, method, etc.) includes the listed elements (e.g., units of a device, components of a composition, substantial steps of a method, etc.), but does not exclude other elements. For example, a composition or article containing a plurality of elements is not limited to only those elements listed herein, but may also include other elements that are not explicitly listed but are generally inherent to the composition or article. Unless expressly stated to the contrary, the term "or" refers to an inclusive "or" rather than an exclusive "or." As used herein, the term "consisting essentially of" when used to define compositions and methods means excluding other elements that have any substantial effect on the combination for the purpose being described, but does not exclude other elements that do not substantially affect the basic and novel characteristics of the invention. As used herein, closed conjunctions such as "consisting of" refer to combinations (units, components, substantial steps, etc.) that exclude other elements, but do not mean to exclude trace amounts of unavoidable impurities unless otherwise specified. Embodiments defined by each of these transitional terms are within the scope of the present invention. Disclosure of a technical solution including the terms "comprising," "including," "having," "containing," or any other similar terms as specific embodiments thereof shall also be deemed to simultaneously disclose corresponding technical solutions including the terms "substantially consisting of" and "consisting of." Herein, all features or conditions defined in the form of numerical ranges or percentage ranges are intended for simplicity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered to have encompassed and specifically disclosed all possible subranges and individual values ​​within the range, particularly integer values. For example, a range description of "1 to 8" should be considered to have specifically disclosed all subranges such as 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8 ... and so on, particularly subranges defined by all integer values, and should be considered to have specifically disclosed individual values ​​within the range such as 1, 2, 3, 4, 5, 6, 7, 8. Similarly, a range description of "between 1 and 8" should be considered to have specifically disclosed all ranges such as 1 to 8, 1 to 7, 2 to 8, 2 to 6, 3 to 6, 4 to 8, 3 to 8, and so on, including their endpoints. Unless otherwise indicated, this interpretation method applies to all contents of the present invention, regardless of whether the scope is extensive or not. As used herein, the terms "expression vector," "expression construct," or "expression cassette" refer to a nucleic acid comprising a nucleotide sequence, such as a coding sequence and a template sequence, as well as sequences necessary for expression of the coding sequence. An expression vector can be viral or non-viral. For example, an expression vector can comprise a nucleic acid construct that, when introduced into a host cell, results in the transcription and / or translation of an RNA or polypeptide, respectively. This definition explicitly includes antisense constructs or sense constructs that are not translated or cannot be translated. One skilled in the art will recognize that the inserted polynucleotide sequence need not be identical to the gene sequence from which it is derived but may only be substantially similar. As used herein, the term "plasmid" generally refers to a non-viral expression vector, such as a nucleic acid molecule encoding a gene and / or regulatory elements necessary for gene expression. As used herein, the term "viral vector" generally refers to a virus-derived nucleic acid capable of transporting another nucleic acid into a cell. When a viral vector is present in a suitable environment, it is capable of directing the expression of one or more proteins encoded by one or more genes carried by the vector. Implementation Method On the one hand, the use of the CRB1 gene in the preparation of a medicament for treating a disease is provided, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. On the one hand, a method for treating a disease is provided, comprising administering CRB1 gene therapy, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. On the one hand, a CRB1 gene therapy drug for treating a disease is provided, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. On the one hand, the use of the CRB1 gene therapy drug in treating a disease is provided, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. On the one hand, the use of the CRB1 gene therapy drug in treating a disease is provided, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. On the one hand, the use of the CRB1 gene therapy drug in the preparation of a medicament for treating a disease is provided, wherein the disease is an intestinal disease and / or an eye disease, in particular an intestinal disease. In some embodiments, administering CRB1 gene therapy comprises administering a CRB1 gene therapy drug. In some embodiments, the CRB1 gene therapy drug comprises genetic material associated with the CRB1 gene. In some embodiments, the genetic material is a nucleotide or peptide, particularly DNA, RNA, or a peptide. In some embodiments, the CRB1 gene therapy drug comprises a CRB1 expression vector. In some embodiments, the CRB1 expression vector comprises a CRB1 target gene. In some embodiments, the CRB1 gene therapy drug comprises a CRB1 target gene. In some embodiments, the CRB1 gene therapy drug comprises a viral vector or a non-viral vector. In some embodiments, the CRB1 gene therapy agent comprises a viral vector. In some embodiments, the viral vector is a DNA virus or an RNA virus. In some embodiments, the viral vector is replication-defective, replication-deficient, replication-attenuated, or replication-competent. In some embodiments, the viral vector comprises a viral vector based on one or more of the following viruses: an adenovirus (e.g., a human adenovirus (e.g., adenovirus serotype 5 (Ad5), adenovirus serotype 26 (Ad26), adenovirus serotype 34 (Ad34), adenovirus serotype 35 (Ad35), or adenovirus serotype 48 (Ad48)), a chimpanzee adenovirus (e.g., ChAd3, ChAd5, ChAd6, ChAd7, ChAd8, ChAd9, ChAd10, ChAd11, ChAd17, ChAd16, ChAd19, ChAd20, ChAd22, ChAd24, ChAdY25, ChAd26, ChAd28, ChAd30, ChAd31, ChAd37, ChAd38, ChAd43, ChAd44, ChAd55, ChAd 63, ChAd63, ChAd68, ChAd73, ChAd82, ChAd83, ChAd143, ChAd144, ChAd145, or ChAd147), gorilla adenovirus (e.g., GC44, GC45, or GC46), rhesus adenovirus (e.g., RhAd51, RhAd52, RhAd53, RhAd54, RhAd55, RhAd56, RhAd57, RhAd58, RhAd59, RhAd60, RhAd61, RhAd62, RhAd63, RhAd64, RhAd65, or RhAd66), bovine adenovirus, canine adenovirus, or porcine adenovirus), adeno-associated virus (e.g., AAV1, AAV2, AAV3, AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV8.2, AAV9, AAVrh10, AAV10, AAV11, AAV2 / 8, AAV2 / 5, AAV2 / 6, or AAV2 / 9), a retrovirus (e.g., an alpha retrovirus (e.g., Rous sarcoma virus), a beta retrovirus (e.g., mouse mammary tumor virus), a gamma retrovirus (e.g., Moloney murine leukemia virus, Moloney murine sarcoma virus, Harvey murine sarcoma virus, Gibbon ape leukemia virus, feline leukemia virus, Freund murine leukemia virus, or murine stem cell virus), a delta retrovirus, an epsilon retrovirus, a lentivirus (e.g., human immunodeficiency virus (type 1 or type 2), Maedi-Vesna virus, caprine arthritis encephalitis virus, equine infectious anemia virus, feline immunodeficiency virus, bovine immunodeficiency virus, or simian immunodeficiency virus), or a foamy virus), an orthomyxovirus (e.g., influenza virus (e.g., influenza A virus, influenza B virus virus, influenza C virus, or influenza D virus)), paramyxovirus (e.g., Newcastle disease virus, mumps virus, measles virus, Sendai virus, pneumovirus, Henipa virus), poxvirus (e.g., vaccinia virus, vaccinia virus, Ankara vaccinia virus, New York vaccinia virus, vaccinia virus Tiantan strain, canarypox virus, fowlpox virus, or myxoma virus), papillomavirus (e.g., human papillomavirus, or bovine papillomavirus), picornavirus (e.g., enterovirus (e.g., rhinovirus, coxsackie virus, echovirus, or poliovirus) or Seneca virus), alphavirus (e.g., Sindbis virus, Ora virus, Babancon virus, Bama Forest virus, Bebaru virus, Bogi River virus, Kabasu virus, Chikungunya virus, Eastern equine encephalitis virus, Everglades virus, Fort Morgan virus, Geta virus, Highland J virus, Zilagachi virus, Mayaro virus, Me Tri virus, Madberg virus, Moso das Pedras virus, Mukambu virus, Ndumau virus, Ochon virus, Pichunna virus, Rio Negro virus, Ross River virus, salmon pancreatic disease virus, Lushan virus, Semliki Forest virus, Southern elephant seal virus, Tonat virus, Trokala virus, Una virus, Venezuelan equine encephalitis virus, Western equine encephalitis virus, or Vodaro River virus), herpes viruses (such as herpes simplex virus (type 1 or 2), varicella virus, Epstein-Barr virus, or cytomegalovirus), arena viruses (such as lymphocytic choriomeningitis virus, inflammatory mammalian arenavirus, Cali mammalian arenavirus, Guanarito virus, Junin virus, Lassa virus, Louyo virus, Machupo virus, Sabia virus, or Whitewater Arroyo virus), coronavirus (e.g., alphacoronavirus, betacoronavirus, gammacoronavirus, or deltacoronavirus), reovirus (e.g., rotavirus), rhabdovirus (e.g., vesicular stomatitis virus, Maraba virus, rabies virus), flavivirus (e.g., yellow fever virus or Zika virus), polyomavirus (e.g., simian vacuolating virus 40), baculovirus, tobacco mosaic virus, or anellovirus. In some embodiments, the viral vector is an adeno-associated viral vector. In some embodiments, the viral vector is AAV1. In some embodiments, the viral vector is AAV2. In some embodiments, the viral vector is AAV3. In some embodiments, the viral vector is AAV3B. In some embodiments, the viral vector is AAV4. In some embodiments, the viral vector is AAV5. In some embodiments, the viral vector is AAV6. In some embodiments, the viral vector is AAV7. In some embodiments, the viral vector is AAV8. In some embodiments, the viral vector is AAV8.2. In some embodiments, the viral vector is AAV9. In some embodiments, the viral vector is AAVrh10. In some embodiments, the viral vector is AAV10. In some embodiments, the viral vector is AAV11. In some embodiments, the viral vector is AAV2 / 8. In some embodiments, the viral vector is AAV2 / 5. In some embodiments, the viral vector is AAV2 / 6. In some embodiments, the viral vector is AAV2 / 9. In some embodiments, the viral vector is an adenoviral vector. In some embodiments, the viral vector is a retroviral vector. In some embodiments, the viral vector is a lentiviral vector. In some embodiments, the viral vector is a poxvirus vector. In some embodiments, the viral vector is an alphavirus vector. In some embodiments, the viral vector comprises the CRB1 gene of interest. In some embodiments, the viral vector comprises a promoter. In some embodiments, the viral vector comprises an enhancer. In some embodiments, the viral vector comprises a silencer. In some embodiments, the viral vector comprises a boundary element. In some embodiments, the viral vector comprises an insulator. In some embodiments, the CRB1 gene therapy agent comprises a non-viral vector. In some embodiments, the CRB1 gene therapy agent comprises a plasmid, a minicircle, a bacterial artificial chromosome, a yeast artificial chromosome, a bacteriophage, a phagemid, a cosmid, or a fosmid. In some embodiments, the CRB1 gene therapy agent comprises naked plasmid DNA. In some embodiments, the CRB1 gene therapy drug comprises a liposome carrier. In some embodiments, the liposome comprises a liposome selected from the group consisting of trimethyl-2,3-dioleyloxypropylammonium chloride (DOTMA), trimethyl-2,3-dioleyloxypropylammonium bromide (DOTAP), dimethyl-2,3-dioleyloxypropyl-2-(2-sperminecarboxamido)ethylammonium trifluoroacetate (DOSPA), trimethyldodecylammonium bromide (DTAB), trimethyltetradecylammonium bromide (TTAB), trimethylhexadecylammonium bromide (CTAB), dimethyldioctadecylammonium bromide (DDAB), dimethyl-2-hydroxyethyl-2,3-dioleyloxypropylammonium bromide (DORI), dimethyl-2-hydroxyethyl-2,3-dioleyloxypropylammonium bromide (DORIE), dimethyl-3-hydroxypropyl-2,3-dioleyloxypropylammonium bromide (DORIE-HP), dimethyl-4-hydroxybutyl-2,3-dioleyloxypropylammonium bromide (DORIE-HP), and dimethyl-4-hydroxybutyl-2,3-dioleyloxypropylammonium bromide (DORIE-HP). at least one of the group consisting of: 1,2-dioleoyl-3-succinyl-sn-glycerocholine ester (DOSC), 3β-[N-(N′,N′-dimethylaminoethyl)carbamoyl]cholesterol (DC-Chol), lipid poly-L-lysine (LPLL), stearylamine, dioleoylphosphatidylethanolamine (DOPE), dioleoylphosphatidylcholine (DOPC), and cholesterol. In some embodiments, the CRB1 gene therapy drug comprises a vesicle vector, particularly an extracellular vesicle vector. In some embodiments, the CRB1 gene therapy drug comprises an exosome vector. In some embodiments, the CRB1 gene therapy drug comprises an engineered exosome vector. In some embodiments, the CRB1 gene therapy drug comprises a microsphere carrier. In some embodiments, the microsphere carrier comprises polyethylene microspheres, latex microspheres, starch microspheres, gelatin microspheres, or polysaccharide microspheres. In some embodiments, the administration is to the intestine. In some embodiments, the administration is to the stomach, jejunum, ileum, cecum, colon, or rectum. In some embodiments, the administration is to the stomach. In some embodiments, the administration is to the jejunum. In some embodiments, the administration is to the ileum. In some embodiments, the administration is to the cecum. In some embodiments, the administration is to the colon. In some embodiments, the administration is to the rectum. In some embodiments, the administration is orally or anally. In some embodiments, the drug for treating a disease is a drug for administration to the intestine. In some embodiments, the drug for treating a disease is a drug for administration to the stomach, jejunum, ileum, cecum, colon, or rectum. In some embodiments, the drug for treating a disease is a drug for administration to the stomach. In some embodiments, the drug for treating a disease is a drug for administration to the jejunum. In some embodiments, the drug for treating a disease is a drug for administration to the ileum. In some embodiments, the drug for treating a disease is a drug for administration to the cecum. In some embodiments, the drug for treating a disease is a drug for administration to the colon. In some embodiments, the drug for treating a disease is a drug for administration to the rectum. In some embodiments, the drug for treating a disease is a drug for administration orally or anally. In some embodiments, the drug for treating the disease is at least one selected from tablets (e.g., sugar-coated tablets, film-coated tablets, sublingual tablets, orally disintegrating tablets, or buccal tablets), pills, powders, granules, capsules (e.g., soft capsules, or microcapsules), lozenges, syrups, solutions, emulsions, suspensions, controlled-release preparations (e.g., instant-release preparations, sustained-release preparations, or sustained-release microcapsules), aerosols, or films (e.g., orally disintegrating films, or oral mucosal adhesive films). In some embodiments, the drug for treating a disease includes a sweetener (e.g., sucrose, xylitol, oligofructose, cyclamate, stevia, or aspartame), a flavoring agent (e.g., spices or essences), a mucilage agent (e.g., sodium alginate, gum arabic, gelatin, methylcellulose, or sodium carboxymethylcellulose), a clarifying agent (e.g., chitosan or gelatin), a preservative (e.g., benzoic acid and its salts, sorbic acid and its salts, or parabens), a disintegrant (e.g., low-substituted hydroxypropyl cellulose, crospovidone, sodium starch glycolate, cross-linked sodium carboxymethyl cellulose, or starch), a binder (e.g., hydroxypropyl cellulose, hypromellose, etc.), a preservative (e.g., benzoic acid and its salts, sorbic acid and its salts, or parabens), a disintegrant (e.g., low-substituted hydroxypropyl cellulose, crospovidone, sodium starch glycolate, cross-linked sodium carboxymethyl cellulose, or starch), a binder (e.g., hydroxypropyl cellulose, , povidone, copovidone, or pregelatinized starch), lubricants (such as stearic acid, magnesium stearate, or sodium fumayl stearate), wetting agents (such as polyoxyethylene sorbitan fatty acid esters, poloxamers, or polyoxyethylene castor oil derivatives), suspending agents (such as hydroxypropyl methylcellulose, hydroxypropyl cellulose, povidone, copovidone, sodium carboxymethylcellulose, or methylcellulose), stabilizers (such as citric acid, fumaric acid, or succinic acid), and fillers (such as starch, sucrose, lactose, or microcrystalline cellulose), binders (such as cellulose derivatives, alginates, gelatin, or polyvinyl pyrrolidone). In some embodiments, the intestinal disease is inflammatory bowel disease (IBD) (i.e., enteritis) and / or intestinal epithelial barrier damage. In some embodiments, the intestinal disease is inflammatory bowel disease. In some embodiments, the inflammatory bowel disease is Crohn's disease or ulcerative colitis. In some embodiments, the intestinal disease is intestinal epithelial barrier damage. In some embodiments, the intestinal disease is an intestinal disease of the colon or an intestinal disease of the rectum. In some embodiments, the intestinal disease is an intestinal disease of the colon. In some embodiments, the intestinal disease is an intestinal disease of the rectum. In some embodiments, the intestinal disease is an inflammatory bowel disease of the colon or an inflammatory bowel disease of the rectum. In some embodiments, the eye disease is ocular inflammation. In some embodiments, the eye disease is optic neuritis, uveitis, conjunctivitis, vitritis, choroiditis, retinitis, retinal vasculitis, sympathetic ophthalmia, retinopathy, glaucoma, cataract, macular degeneration (e.g., age-related macular degeneration), Behçet's disease, or Vogt-Koyanagi-Harada syndrome. In some embodiments, the eye disease is an optic nerve disease, in particular glaucoma, optic neuritis (e.g., papillitis or retrobulbar neuritis), ischemic optic neuropathy (e.g., non-arteritic ischemic optic neuropathy or arteritic ischemic optic neuropathy), inflammatory optic neuropathy, optic disc edema, optic atrophy, or optic disc vasculitis. In some embodiments, the eye disease is a retinal disease, such as retinal tear, retinal detachment, retinopathy (e.g., diabetic retinopathy), epiretinal membrane, macular hole, macular degeneration, or retinitis pigmentosa. In some embodiments, the intestinal disease and / or eye disease, in particular the intestinal disease, is an intestinal disease and / or eye disease, in particular an intestinal disease, associated with the CRB1 gene, in particular a mutation in the CRB1 gene. In some embodiments, the intestinal disease and / or eye disease, in particular the intestinal disease, is an intestinal disease and / or eye disease, in particular an intestinal disease, associated with a Rd8 mutation in the CRB1 gene. In some embodiments, the intestinal disease and / or eye disease, in particular the intestinal disease, is also an intestinal disease and / or eye disease associated with at least one gene selected from the group consisting of ABCA4, ABCC6, ABCC9, ACBD5, ACO2, ACO2, ACTG1, ADGRV1, AHI1, AIPL1, ALMS1, AMY2B, APC, ARFGEF1, ARL13B, ARL13B, ARL6, ARMC9, ATOH7, B9D1, BAG3, BBS1, BBS1, BBS2, BBS5, BEST1, C2CD3 , CA4, CABP4, CACNA1F, CBS, CC2D2A, CDH23, CDH23, CDHR1, CEMIP2, CEP104, CEP250, CEP290, CEP290, CEP41, CEP78, CERKL, CFAP410, CFAP418, CHM, CLCC 1. CLCN7, CLN3, CLN5, CLN8, CLRN1, CLRN1, CNGA1, CNGA1, CNGA3, CNGB1, CNGB3, CNNM4, COL11A1, COL11A2, COL18A1, COL2A1, COL4A1, COL9A1, COL9A2, CP , CP, CPLANE1, CRB1, ERCC4, CSPP1, CTNNA1, CYP4V2, DHDDS, DYNC2H1, DYNC2I1, DYNC2I2, ENPP1, ERCC4, EVC2, EYS, EYS, F5, FAM161A, FBN1, FKRP, FKTN, F LG, FLVCR1, FOXE3, FUZ, GLB1, GMPPB, GNAT1, GRK1, GRM6, GUCA1A, GUCA1B, GUCY2D, HADHA, HGSNAT, HPS3, HPS5, IDH3B, IFT122, IFT140, IFT140, IFT43, I FT52, IFT74, IFT80, IFT80, IFT81, IFT88, IKBKG, IMPDH1, IMPG2, INPP5E, INTU, IQCB1, IQCE, IREB2, KCNJ13, KCNQ1, KCNV2, KIAA0586, KIAA0753, KIF7, KIZ, KIZ-AS1, KLHL7, KRIT1, LBR, LCA5, LOC101927157, LOC111365204, LRP2, LRP5, MAK, MAPKAPK3, MATK, MCOLN1, MERTK, MKS1, MPDZ, MT-ATP6, MT-CO3,MT-TE, MT-TL1, MTHFR, MUTYH, MYO7A, MYO7A, NMNAT1, NPHP1, NR2E3, OCA2, OTX2, PANK2, PAX6, PCARE, PC DH15, PDE6A, PDE6B, PDE6B, PDE6D, PEX1, PEX1, PEX12, PEX26, PEX6, PHF3, PITPNM3, PKD2, PLA2G5, POC5 , POMT1, PRCD, PRDM13, PROM1, PRPF3, PRPF31, PRPF8, PRPH2, RAD51C, RBP3, RBP4, RD3, RDH12, RDH5, RGR , RGR, RHO, RIMS1, RLBP1, ROM1, RP1, RP1L1, RP2, RPE65, RPE65, RPGR, RPGRIP1, RPGRIP1L, RS1, SACS, SAG ,SCAPER,SDCCAG8,SIX6,SLC19A1,SLC22A5,SLC26A4,SLC2A9,SLTM,SNRNP200,SPAG17,SPATA7,SPG11 , TFAP2A, TGFB2, TGFBR2, TMEM107, TMEM237, TMEM67, TOGARAM1, TOPORS, TPP1, TRAF3IP1, TREX1, TRIM5 9-IFT80, TSPAN12, TTC21B, TTC21B, TTC8, TULP1, USH1C, USH2A, USH2A, USH2A, USH2A, USH2A-AS1, VAC1 4. VCAN, VCAN, VCAN-AS1, VHL, VPS13B, WDR19, WDR19, WDR35, WDR73, YARS1, ZFYVE26, ZFYVE26, and ZNF408. , Example Experimental methods mice C57BL / 6N mice carrying the Rd8 mutation (Crb1 rd8 / Rd8 , named Rd8 mice) and C57BL / 6J mice (Crb1 wt / wt WT mice (both WT and WT mice) were purchased from Jicui Yaokang Biotechnology Co., Ltd. and maintained in a standard specific pathogen-free animal facility at the Zhongshan Eye Center of Sun Yat-sen University. Rd8 mice were purified at the animal facility of Huazhong Agricultural University. Purified germ-free mice were maintained in a sterile state, and fecal samples were tested for microorganisms and parasites weekly to ensure sterility. Genotyping of Rd8 and Crb1-KO mutations was performed according to Mattapallil et al., 2012 or according to the instructions of Jicui Pharmaceutical Biotechnology Co., Ltd. All animal experiments were reviewed and approved by the Institutional Animal Care and Use Committee (IACUC) of the Zhongshan Eye Center of Sun Yat-sen University and Huazhong Agricultural University and adhered to the relevant sections of the ARVO Statement for the Use of Animals in Research. Histology Mice were sacrificed by cervical dislocation, and the desired tissues were removed and fixed with 4% paraformaldehyde (PFA) at 4°C for 24 h. Samples were washed three times with PBS, embedded in paraffin, and sectioned into 10 μm slices using a microtome. Sections were stained with hematoxylin and eosin (H&E). Immunofluorescence Mice were perfused with 4% PFA via cardiac puncture. Eyes were removed and post-fixed for 15 minutes at room temperature. Colonic tissue was cut into approximately 5 mm pieces and post-fixed in 4% PFA for 4 hours at room temperature. Colonic and cecal tissues were washed three times with PBS for 15 minutes, incubated overnight in 30% sucrose / PBS, embedded in OCT, and cryosectioned. Frozen tissue sections (12 μm) were incubated with blocking buffer (10% donkey serum, 0.1% Triton X-100 in PBS) for 30 minutes at room temperature, incubated with primary antibodies diluted in blocking buffer overnight at 4°C, and incubated with fluorochrome-conjugated secondary antibodies or phalloidin for 2 hours at room temperature, with extensive washing after each incubation. Fluorescence microscopy images were acquired using a Zeiss LSM880 confocal microscope. The primary antibodies used included anti-CRB1 (PA5-66373, ThermoFisher; 1:50), anti-IBA1 (ab178846, Abcam; 1:500), anti-CD4 (60-0042-U100, Tonbo Bioscience; 1:100), anti-ZO-1 (61-7300, ThermoFisher; 1:500), anti-occludin (33-1500, ThermoFisher; 1:200), AlexaFluor 488-Phallodin (A12379, ThermoFisher; 1:500), anti-β-Catenin (Cell Signaling Technology, 8480; 1:200), and anti-GFP (ab290, Abcam; 1:500). The specificity of these antibodies was verified before immunofluorescence staining. Quantitative real-time polymerase chain reaction (qPCR) Fresh mouse colon tissue (approximately 1 cm) was rapidly frozen in liquid nitrogen, ground, and lysed with RNA extraction and lysis buffer. Total RNA was then purified using the Qiagen RNeasy Plus kit and reverse-transcribed into cDNA using the Takara PrimeScript RT kit with gDNA cleanup enzyme. The expression levels of the corresponding genes were analyzed by qPCR. Data were normalized to β-actin. AAV vector preparation The Crb1 target gene (SEQ ID NO: 1) or the GFP target gene (SEQ ID NO: 2) was cloned into the target AAV expression vector by enzyme digestion-ligation-transformation. Positive clones were obtained and sequenced to confirm the correctness of the inserted fragment. The AAV recombinant vector was transfected into HEK293T cells and the virus was harvested after 72 hours. After purification, concentration and virus titer detection, AAV2 virus particles encoding GFP or Crb1 genes were obtained and used as AAV2 vectors. AAV2 vector delivery Rd8 newborn mice were administered 5×10 11 AAV2 vectors (dissolved in 20 μl 1× PBS) containing GFP or Crb1 genes via the anus at 10–12 days of age. The AAV2 vectors are shown in Figure 5 . During the enema procedure, a small amount of sterile mineral oil was used to lubricate the outside of a 10 μL pipette tip attached to the pipette shaft. The pipette tip was then inserted into the anus and the pipette was gently depressed to release the ultrapure AAV viral solution (in vivo grade). Therapeutic effects of regulating Crb1 expression in the intestine on Rd8 mice Using the AAV2 viral vector system (Figure 1), we obtained Rd8 mice that expressed CRB1 protein at the apical end of intestinal epithelial cells but did not express Crb1 in other tissues including the liver, spleen, or retina (see Figure 2, A–B). These mice were named Rd8-AAV-Crb1 mice. As shown in Figure 2, C–G, expression of CRB1 in intestinal epithelial cells restored the adherens junction integrity of the colonic epithelial barrier (see Figure 2, C–D), whereas changes in colonic microvilli in Rd8 mice remained unchanged (see Figure 2, E–G). Although the structural defects of Bruch's membrane were not restored in Rd8-AAV-Crb1 mice (see Figure 2, H–I), intestinal permeability (see Figure 2, J) and bacterial translocation in peripheral blood (see Figure 2, K–L) were significantly reduced in Rd8-AAV-Crb1 mice. The results showed that restoring CRB1 expression in the intestine, for example by administering CRB1 gene therapy drugs such as viral vectors to mice, could improve the morphological defects of the colon epithelial barrier in mice, thereby treating intestinal diseases. The above-mentioned embodiments are only preferred embodiments of the present application and cannot be used to limit the scope of protection of the present application. Any non-substantial changes and replacements made by technicians in this field based on the present application shall fall within the scope of protection required by the present application.

Claims

1. Use of the CRB1 gene in the preparation of a drug for treating a disease, wherein the disease is an intestinal disease.

2. Use of CRB1 gene therapy drugs in the preparation of drugs for treating diseases, wherein the diseases are intestinal diseases.

3. The use according to claim 2, wherein The CRB1 gene therapy drug includes a viral vector or a non-viral vector; Optionally, the CRB1 gene therapy drug includes a non-viral vector; Optionally, the CRB1 gene therapy drug comprises a plasmid, a minicircle, a bacterial artificial chromosome, a yeast artificial chromosome, a bacteriophage, a phagemid, a cosmid, or a F-cosmid; Optionally, the CRB1 gene therapy drug comprises a liposome carrier; Optionally, the CRB1 gene therapy drug includes a microsphere carrier.

4. The use according to claim 2 or 3, wherein The CRB1 gene therapy drug includes a viral vector; Optionally, the viral vector comprises a viral vector based on one or more of the following viruses: an adenovirus, an adeno-associated virus, a retrovirus, an orthomyxovirus, a paramyxovirus, a poxvirus, a papillomavirus, a picornavirus, an alphavirus, a herpesvirus, an arenavirus, a coronavirus, a reovirus, a rhabdovirus, a flavivirus, a polyomavirus, a baculovirus, a tobacco mosaic virus, or an anellovirus.

5. The use according to any one of claims 2 to 4, wherein The CRB1 gene therapy drug includes an adeno-associated virus vector; Optionally, the viral vector includes a viral vector based on one or more of the following viruses: AAV1, AAV2, AAV3, AAV3B, AAV4, AAV5, AAV6, AAV7, AAV8, AAV8.2, AAV9, AAVrh10, AAV10, AAV11, AAV2 / 8, AAV2 / 5, AAV2 / 6 or AAV2 / 9.

6. The use according to any one of claims 2 to 5, wherein The CRB1 gene therapy drug includes a CRB1 expression vector; Optionally, the CRB1 expression vector includes a CRB1 target gene.

7. The use according to any one of claims 1 to 6, wherein The drug for treating the disease is a drug for administration to the intestine; Optionally, the drug for treating the disease is a drug administered to the colon or rectum; Optionally, the drug for treating a disease is a drug administered to the colon.

8. The use according to any one of claims 1 to 7, wherein The intestinal disease is inflammatory bowel disease and / or intestinal epithelial barrier damage; Optionally, the intestinal disease is inflammatory bowel disease; Optionally, the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

9. The use according to any one of claims 1 to 8, wherein The intestinal disease is an intestinal disease of the colon or rectum.

10. The use according to any one of claims 1 to 9, wherein The intestinal disease is an intestinal disease related to the CRB1 gene.

11. Use of CRB1 gene and / or CRB1 gene therapeutic drug in the preparation of a drug for treating a disease, wherein the disease is an eye disease.

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