Application of RBFOX2 and related substances thereof in treatment of colitis

Abstract

The invention relates to the technical field of biomedical engineering, provides an application of an RBFOX2 inhibitor in treatment of inflammatory bowel diseases, and particularly relates to an effect of interfering RNA or antisense oligonucleotide or recombinant expression vector of a targeted RBFOX2 coding sequence in preparation of products for treating colitis and related symptoms and / or other complications thereof. The invention also relates to a corresponding recombinant vector or pharmaceutical composition. The RBFOX2 inhibitor can effectively reduce the level of pro-inflammatory cytokines and inhibit the activation of macrophages, has effective application values of relieving and treating colitis and other symptoms and / or other complications of the inflammatory bowel disease, provides a new target spot for the treatment of the inflammatory bowel disease, and has a wide clinical application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to the use of RNA binding fox-1 homolog 2 (RBFOX2) as a target in the preparation of drugs for treating inflammatory bowel disease (IBD), and also to a carrier and a pharmaceutical composition containing the same. Background Art

[0002] Inflammatory bowel disease (IBD) is a group of chronic, recurrent, inflammatory intestinal disease states, mainly including Crohn's disease (CD) and ulcerative colitis (UC). These two IBDs have both overlapping and differences in clinical and pathological characteristics. In recent years, the incidence and prevalence of IBD in my country have shown a significant upward trend. At present, the pathogenesis of IBD is not fully understood. Previous studies have shown that the interaction between intestinal microorganisms, intestinal epithelial cells and innate immunity is widely involved in the pathogenesis of IBD. The innate immune response plays an important role in this. In the intestines of healthy people, macrophages do not produce proinflammatory cytokines, but still have phagocytic and antibacterial activity. Studies have shown that in CD patients, CD14 + Macrophages express marker molecules of macrophages (CD14, CD33, and CD68) and dendritic cells (CD205 and CD209) at the same time, and can produce a large amount of inflammatory factors IL-6, IL-23, and TNF, promoting the production of IFN-γ. Although IBD treatment drugs have made rapid progress in recent years, they all target a single inflammatory factor or inflammatory pathway, resulting in limited clinical efficacy. Therefore, by studying the pathological process and pathogenic genes of IBD, fundamentally blocking and intervening in the pathological effects of abnormal activation of immune cells in the colonic inflammatory microenvironment has important practical significance and clinical transformation prospects.

[0003] By analyzing the differentially expressed genes in the synovial tissues of IBD patients and healthy subjects, it was identified that the high expression of RNA binding fox-1 homolog 2 (RBFOX2) in peripheral blood mononuclear cells (PBMCs) of IBD patients may be involved in the occurrence and development of IBD. RBFOX2, also known as RBM9 or FOX2, is 390 amino acids in length and is highly conserved between yeast and a variety of mammals, suggesting that this protein has extremely important biological functions. RBFOX2 has been reported to play an important biological role in a variety of tumors such as diabetes, heart failure, fatty liver, pancreatic cancer, and myeloid leukemia. Currently, there are few studies on this protein, mainly focusing on the formation of its classic spliceosome catalytic center, that is, RBFOX2 can perform alternative splicing of pre-mRNA by recognizing the evolutionarily conserved GCAUG motif. Studies have found that in the pancreas, RBFOX2 can regulate the selective splicing of genes required for insulin granule docking and exocytosis, thereby affecting insulin secretion; in cardiomyocytes, RBFOX2 can splice multiple cGMP-PKG-Ca2 + RBFOX2 is a key gene in the pathway, causing a significant increase in calcium ion levels in cardiomyocytes; in liver cells, RBFOX2 regulates the alternative splicing of the class B type I scavenger receptor Scarb1 to maintain cholesterol homeostasis in the liver lipogenesis environment; transforming growth factor TGF-β can induce RBFOX2 to catalyze and mediate TAK1 alternative splicing, playing a role in tumor epithelial-mesenchymal transition and chemotherapy drug resistance. In addition, RBFOX2 can also mediate H3K27me3-mediated polycomb complex 2 (PRC2), mediate N6-methyladenosine methylation (m 6 A) directly binds to the RNA binding motif 15 (RBM15) of RBFOX2 and affects gene transcription. Therefore, inhibiting the activity of RBFOX2 has a potential therapeutic effect on the above diseases. Although the role of RBFOX2 protein in autoimmune diseases such as IBD has not been reported, we reasonably speculate that high expression of RBFOX2 may be related to the occurrence and development of IBD.

[0004] In summary, there is an urgent need in this field to develop new strategies for the relief and treatment of IBD, to inhibit the activation and pathogenic effects of immune cells in the local inflammatory microenvironment of the colon, and to control the resulting IBD and its related symptoms and / or other complications. Summary of the invention

[0005] The present invention is aimed at the above problems. First, the adverse effects of abnormal expression of RBFOX2 on IBD are confirmed. Then, based on this target, the recovery of colon inflammatory damage is observed after the abnormal expression of RBFOX2 is eliminated or alleviated, thereby determining the role of RBFOX2 in the onset and progression of IBD.

[0006] The research process of the present invention is as follows:

[0007] Using the Cre-Loxp system to knock out RBFOX2 in mouse myeloid cells, it was found that RBFOX2 knockout alleviated the inflammatory response of the colon in the mouse experimental IBD model, including intestinal villus damage and pathogenic immune cell infiltration. By analyzing the expression of related factors in macrophages after RBFOX2 knockout, it was found that CXC chemokine ligand 1 (CXCL1) is a key chemokine controlled by RBFOX2 and is involved in the RBFOX2 deficiency-mediated IBD disease remission process. Silencing the expression of RBFOX2 using specific small interfering RNA (siRNA) targeting the RBFOX2 gene can also downregulate the production of CXCL1 in mouse peritoneal macrophages. Based on this, it is concluded that the high expression of RBFOX2 in IBD plays an important role in the onset and progression of IBD by upregulating the production of CXCL1 and recruiting more immune inflammatory cells to infiltrate.

[0008] Based on the above research, the specific technical solutions of the present invention are as follows:

[0009] The first aspect of the present invention provides use of a RBFOX2 inhibitor in the preparation of a medicament for treating IBD or its complications.

[0010] IBD is an inflammation of the colon caused by the pathogenicity of macrophages and T lymphocytes. Symptoms associated with IBD are colitis-related symptoms characterized by abdominal pain, diarrhea, bloody stools, etc., which can lead to malnutrition in severe cases. In addition, IBD can also affect other parts of the body, such as joints, eyes, mouth, liver, gallbladder, and skin. IBD also increases the risk of cancer in affected intestinal areas.

[0011] Preferably, the RBFOX2 inhibitor is an interfering RNA or antisense oligonucleotide targeting the RBFOX2 coding sequence. The above substances can target RBFOX2 alone, or can be a compound targeting RBFOX2 in the form of liposomes or nanoparticles.

[0012] Wherein, the RBFOX2 coding sequence is selected from any of the following situations:

[0013] (i) the nucleotide sequence encoding the RBFOX2 protein as shown in SEQ ID NO.1;

[0014] (ii) a sequence homologous to the RNA in (i);

[0015] (iii) A molecule in which one or more nucleotides are substituted, deleted or added in the RNA sequence of (i) and / or (ii), and which encodes a protein or polypeptide having the effect of aggravating IBD symptoms.

[0016] The RBFOX2 protein is selected from any of the following:

[0017] (a) the amino acid sequence shown in SEQ ID NO.2;

[0018] (b) homologous to or having sequence homology with the amino acid sequence of SEQ ID NO. 2 (e.g., more than 80% homologous or having more than 80% sequence identity, such as 80%, 85%, 90%, 95%, 98%, 99%), and having a

[0019] Proteins or peptides that have an effect on IBD and its related symptoms and / or other complications;

[0020] (c) A protein or polypeptide derived from (a) or (b) in which one or more amino acids are substituted, deleted or added in the amino acid sequence of (a) or (b) and which has the effect of aggravating IBD symptoms.

[0021] The interfering RNA or antisense oligonucleotide sequence targeting the RBFOX2 coding sequence is shown in any of the following:

[0022] siRNA 132213-1:AACTCCTGACGCAATGGGTTCA (SEQ ID NO.3);

[0023] siRNA 132214-1: CCAGACCAGTGAACATAACCT (SEQ ID NO.4);

[0024] siRNA 132215-1: GGAGAACAGAGTAGCAATTCA (SEQ ID NO. 5).

[0025] The second aspect of the present invention provides a recombinant vector targeting RBFOX2, comprising an expression vector and an interfering RNA or antisense oligonucleotide targeting the RBFOX2 coding sequence inserted into the expression vector, the sequence of which is as shown above.

[0026] The third aspect of the present invention provides use of a recombinant vector targeting RBFOX2 in the preparation of a drug for treating IBD.

[0027] The vectors include viral vectors and non-viral vectors.

[0028] The "viral vector" includes adeno-associated virus and adenovirus. Suitable viral vectors are well known to those of ordinary skill in the art.

[0029] The "non-viral vector" includes liposomes or lipid complexes, cationic polymers, chitosan polymers and nanoparticle vectors, etc. Suitable non-viral vectors are well known to those skilled in the art.

[0030] In a fourth aspect, the present invention provides a pharmaceutical composition for treating IBD and its complications, comprising an active ingredient and a medically acceptable excipient, carrier or diluent, wherein the active ingredient is the above-mentioned RBFOX2 inhibitor or a recombinant vector targeting RBFOX2.

[0031] The recombinant virus of the present invention and pharmaceutically acceptable excipients together constitute a pharmaceutical composition for treating IBD, thereby exerting therapeutic effects more stably.

[0032] In terms of drug form, the drug or drug composition is suitable for administration by a method selected from the following group: oral administration, injection (such as direct naked DNA or protein injection, liposome-encapsulated DNA or mRNA injection), gold-coated gene gun bombardment, replication-defective adenovirus carrying target DNA or protein encoded by the target gene, enema administration, and transdermal administration.

[0033] Generally, liquid preparations can be kept stable for at least one year at 2°C-8°C, and freeze-dried preparations can be kept stable for at least six months at 30°C. The preparations can be suspensions, water injections, freeze-dried preparations and other preparations commonly used in the pharmaceutical field.

[0034] When the composition of the present invention is administered to animals including humans, the dosage varies depending on the patient's age and weight, disease characteristics and severity, and administration route. The results of animal experiments and various circumstances can be referred to, and the total dosage cannot exceed a certain range.

[0035] Furthermore, the pharmaceutical composition of the present invention can be used in combination with other pharmaceutical compositions for treating IBD-related symptoms or other complications.

[0036] A fifth aspect of the present invention provides use of a reagent for detecting the expression level of RBFOX2 in preparing an IBD detection kit or a drug screening kit.

[0037] The experimental results showed that the expression of RBFOX2 was significantly upregulated in IBD patients, and RBFOX2 knockout alleviated the occurrence and development of DSS-induced colitis in mice, inhibited the activation, infiltration and tissue damage of inflammatory cells in the colon microenvironment, suggesting that RBFOX2 is a promoter of IBD and has the effect of promoting the occurrence and development of IBD. Based on this, it can be inferred that RBFOX2 can be used as a diagnostic marker or drug screening marker for IBD.

[0038] The method for drug screening using RBFOX2 as a marker is as follows:

[0039] (A) treating cells, tissues or animals with IBD or mimicking IBD symptoms or other inflammatory environments with a candidate substance;

[0040] (B) detecting the level of RBFOX2 or a nucleic acid molecule encoding the protein in the cell, tissue or animal;

[0041] (C) After treatment with the candidate substance, if the level of RBFOX2 or the nucleic acid molecule encoding the protein is lower than the RBFOX2 expression level in the IBD model tissue or cell, it indicates that the candidate substance has the effect of treating IBD by inhibiting RBFOX2.

[0042] In a sixth aspect of the present invention, an IBD diagnostic kit or drug screening kit is provided, which comprises a reagent for detecting the expression level of RBFOX2 in a biological sample, and is composed of a reverse transcription system, a primer system, and an amplification system. Diagnosis or drug screening is achieved by detecting the nucleic acid expression level of RBFOX2.

[0043] Among them, the primer sequences of the primer system are as follows:

[0044] Rbfox2-F1: 5'-AACCAGGAGCCAACAACAACT-3' (SEQ ID NO. 6);

[0045] Rbfox2-R1: 5'-GAACGGGATGGTAGTAAAAGGC-3' (SEQ ID NO.7);

[0046] Rbfox2-F2: 5'-TCCGAGGAGCCATCTGAGG-3' (SEQ ID NO.8);

[0047] Rbfox2-R2: 5'-AATCCGTCCTGGTAAACCACA-3' (SEQ ID NO. 9).

[0048] Beneficial protection and effects of the present invention:

[0049] The present invention provides the use of interfering RNA or antisense oligonucleotide of RBFOX2 coding sequence in the preparation of a drug for treating IBD or its complications. Experiments have confirmed that RBFOX2 knockout inhibits the occurrence and development of DSS-induced colitis in mice, reduces inflammatory cell activation, infiltration and tissue damage in the colon microenvironment, indicating that RBFOX2 is a pathogenic factor of IBD and has the effect of aggravating immune inflammation and colon pathological damage. Therefore, the present invention provides a new target for the treatment of IBD by targeting the inhibition of RBFOX2, and has broad clinical application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0050] Figure 1Disease severity scores and weight loss in the DSS-induced colitis model of RBFOX2 knockout mice (CKO mice); A, disease severity scores of WT and cKO mice; B, weight loss ratio of WT and cKO mice.

[0051] Figure 2 It shows that RBFOX2 knockout reduces the inflammatory response and tissue damage in the colon microenvironment; A, eosin & hematoxylin staining results of colon tissue after DSS-induced colitis model in WT and cKO mice; B, histopathological scores of WT and cKO mice.

[0052] Figure 3 The results show that knocking out or interfering with the expression of RBFOX2 reduced the expression level of CXCL1 in mouse peritoneal macrophages; A, the mRNA expression level of CXCL1 in peritoneal macrophages of mice with RBFOX2 knocked out; B, the mRNA expression level of CXCL1 in peritoneal macrophages of mice with RBFOX2 interfered; C, the expression level of CXCL1 in the supernatant of peritoneal macrophages of mice with RBFOX2 knocked out.

[0053] Figure 4 The results showed that AAV9 virus interfering with RBFOX2 expression can inhibit the expression level of CXCL1 in macrophages and pathological damage of colon tissue; A, immunoblotting results of detecting RBFOX2 protein expression after macrophages were infected with recombinant virus AAV9-RBFOX2-RNAi; B, recombinant virus AAV9-RBFOX2-RNAi inhibited the production of CXCL1 by mouse peritoneal macrophages; C, eosin & hematoxylin staining results of colon tissue after treatment of IL10- / - mice with spontaneous colitis with recombinant virus AAV9-RBFOX2-RNAi through tail vein injection. DETAILED DESCRIPTION

[0054] The following examples and experimental examples further illustrate the present invention and should not be construed as limiting the present invention. The examples do not include a detailed description of conventional methods, such as PCR methods, methods for constructing vectors and plasmids, methods for inserting protein-encoding genes into such vectors and plasmids, or methods for introducing plasmids into host cells. Such methods are well known to those skilled in the art and are described in many publications, including Sambrook, J., Fritsch, E. F. and Maniais, T. (1989) Molecular Cloning: A Laboratory Manual, 2 nd edition, Cold spring Harbor Laboratory Press.

[0055] Unless otherwise specified, percentages and parts are calculated by volume. Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention, and the preferred implementation methods and materials described in the specific implementation methods are for demonstration purposes only.

[0056] Example 1: RBFOX2 knockout alleviates clinical phenotypes in a mouse DSS-induced colitis model

[0057] Construction of RBFOX2 myeloid cell conditional knockout mice. fl / fl The mouse was constructed by Saiye Biotechnology Co., Ltd. and then hybridized with Lyz2-Cre mice to obtain RBFOX2 fl / fl Lyz2-Cre + / – mice, called cKO mice, to achieve specific knockout of the RBFOX2 gene in myeloid cells, and also obtain control mice RBFOX2 fl / fl Lyz2-Cre – / – (referred to as WT mice). These mice were housed in a special pathogen-free (SPF) environment.

[0058] WT and cKO mice were fasted overnight and given 3% DSS in drinking water to establish a DSS-induced colitis model in mice. Monitoring indicators included weight change (>110% 0 points; 90%-110% 1 point; 70%-89% 2 points; <70% 3 points), stool characteristics (normal 0 points; soft 1 point; loose 2 points; loose stool 3 points) and blood in stool (none 0 points; a little 1 point; obvious 2 points; mainly blood 3 points), with the highest total score being 9 points.

[0059] The results showed that compared with the control group WT mice, the clinical scores of RBFOX2 cKO mice were lower ( Figure 1 A), the degree of weight loss was significantly slowed down ( Figure 1 B).

[0060] The results showed that RBFOX2 knockout alleviated the occurrence and development of DSS-induced colitis in mice, suggesting that RBFOX2 is a pathogenic factor of IBD.

[0061] Example 2: RBFOX2 knockout alleviates inflammatory response and tissue damage in the colon microenvironment

[0062] RBFOX2 myeloid cell conditional knockout mice were constructed (same as in Example 1). The colon of the mice was obtained, fixed with paraformaldehyde, and then stained with eosin and hematoxylin to observe the histopathological changes of the colon of the mice.

[0063] The results showed that the infiltration of pathogenic inflammatory cells in the colon microenvironment of RBFOX2 knockout mice was significantly reduced, and the crypt structure was more complete ( Figure 2 A). The degree of colon tissue pathological damage in RBFOX2 knockout mice was significantly reduced compared with that in WT mice, and the histological score was lower ( Figure 2 B).

[0064] The results showed that RBFOX2 knockout alleviated inflammatory cell activation, infiltration and tissue damage in the colon microenvironment, suggesting the immune-promoting effect of RBFOX2.

[0065] Example 3: Knockout or interference with RBFOX2 expression enhances CXCL1 expression in mouse peritoneal macrophages

[0066] Primary peritoneal macrophages of RBFOX2 WT and cKO mice were obtained and stimulated with lipopolysaccharide (LPS) (100 ng / ml) to simulate the activation state of macrophages in IBD. Total RNA of macrophages was collected and reverse transcription-PCR and real-time quantitative PCR were used to detect CXCL1 levels. The supernatant was collected and ELISA was used to detect CXCL1 protein levels.

[0067] Peritoneal macrophages of wild-type mice were obtained and transfected with siRNA sequences targeting mouse RBFOX2 (sequences are shown in SEQ ID NO.3-5, synthesized by Santa Cruz) and control sequences (both purchased from Santa Cruz). 72 hours after the above transfection, peritoneal macrophages were stimulated with lipopolysaccharide LPS (100 ng / ml) to simulate the activation state of macrophages in IBD, and total RNA of macrophages was collected, and CXCL1 levels were detected by reverse transcription PCR and real-time quantitative fluorescence PCR.

[0068] The specific sequence for constructing siRNA (siRNA-RBFOX2) that specifically interferes with RBFOX2 expression is as follows:

[0069] siRNA 132213-1:AACTCCTGACGCAATGGGTTCA (SEQ ID NO.3);

[0070] siRNA 132214-1: CCAGACCAGTGAACATAACCT (SEQ ID NO.4);

[0071] siRNA 132215-1: GGAGAACAGAGTAGCAATTCA (SEQ ID NO.5);

[0072] The results showed that peritoneal macrophages from mice with RBFOX2 knockout produced lower levels of CXCL1 ( Figure 3 A). The level of CXCL1 produced by peritoneal macrophages in mice with RBFOX2 knockdown was significantly decreased ( Figure 3 B). The CXCL1 level in the supernatant of peritoneal macrophages from mice with RBFOX2 knockout was also significantly decreased ( Figure 3 C).

[0073] The results showed that reducing the expression of RBFOX2 by gene knockout or siRNA interference could inhibit the activation of macrophages and reduce the production of CXCL1 in macrophages.

[0074] Example 4: AAV9 virus interfering with RBFOX2 can inhibit CXCL1 levels in macrophages and colon pathological damage.

[0075] Shanghai JiKai Gene Co., Ltd. was commissioned to construct an adeno-associated virus that produces targeted interference with RBFOX2 expression based on the adeno-associated virus vector of AAV9 according to the siRNA sequence targeting the RBFOX2 gene in the aforementioned Example 3, and it was named AAV9-RBFOX2-RNAi. The above three AAV9-RBFOX2-RNAi and the control empty virus AAV9-control were respectively infected with primary peritoneal macrophages, and the infection efficiency of the virus was detected by immunoblotting to confirm its interference effect. Then, the above viruses were infected in mouse peritoneal macrophages (same as in Example 3), and lipopolysaccharide LPS (100ng / ml) was used for stimulation to simulate the activation state of macrophages in IBD, and the total RNA of macrophages was collected, and the CXCL1 level was detected by real-time quantitative PCR after reverse transcription PCR.

[0076] The results showed that AAV9-RBFOX2-RNAi( Figure 4 Lane A2, 3, 4) compared with AAV9-control ( Figure 4 A1) It can be observed that the expression of RBFOX2 protein was significantly reduced; the third AAV9-RBFOX2-RNAi and AAV9-control were used to infect mouse peritoneal macrophages, and it was found that interfering with the expression of RBFOX2 inhibited the production of CXCL1 in mouse peritoneal macrophages ( Figure 4 B); AAV9-RBFOX2-RNAi and AAV9-control were injected into the tail vein of Il10- / - mice with spontaneous colitis. The results of eosin and hematoxylin staining of colon tissues showed that interference with RBFOX2 expression in vivo significantly reduced colon inflammation damage ( Figure 4 C).

[0077] The results showed that targeted inhibition of RBFOX2 could inhibit the activation of macrophages, reduce the production of CXCL1 in macrophages, and alleviate colonic inflammatory response and pathological damage.

[0078] The nucleotide sequence encoding the RBFOX2 protein in the present invention is shown in SEQ ID NO.1:

[0079]

[0080] The amino acid sequence of the RBFOX2 protein in the present invention is shown in SEQ ID NO.2:

[0081] MEKKKMVTQGNQEPTTTPDAMVQPFTTIPFPPPPQNGIPTEYGVPHTQDYAGQTGEHNLTLYGSTQAHGEQSSNSPSTQNGSLTTEGGAQTDGQQSQTQSSENSESKSTPKRLHVSNIPFRFRDPDLRQMFGQFGKILDVEIIFNERGSKGFGFVTFENSADADRAREKLHGTVVEGRKIEVNNATARVM TNKKMVTPYANGWKLSPVVGAVYGPELYAASSFQADVSLGNDAAVPLSGRGGINTYIPLISLPLVPGFPYPTAATTAAAFRGAHLRGRGRTVYGAVRAVPPTAIPAYPGVVYQDGFYGADLYGGYAAYRYAQPATATAATAAAAAAAAYSDGYGRVYTADPYHALAPAASYGVGAVASLYRGGYSRFAPY

[0082] The preferred embodiments of the present invention have been specifically described above, but the present invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. Use of RBFOX2 inhibitors in the preparation of drugs for treating inflammatory bowel disease or its complications.

2. The use according to claim 1, characterized in that: The inflammatory bowel disease is an inflammation of the colon caused by pathogenic macrophages and T lymphocytes, including ulcerative colitis and Crohn's disease.

3. The use according to claim 1, characterized in that: The RBFOX2 inhibitor is selected from interfering RNA or antisense oligonucleotides targeting the RBFOX2 coding sequence.

4. The use according to claim 3, characterized in that: The sequence of the interfering RNA or antisense oligonucleotide targeting the RBFOX2 coding sequence is shown in any one of SEQ ID NO.3 to SEQ ID NO.

5.

5. A recombinant vector targeting RBFOX2, characterized in that: The invention comprises an expression vector and an interfering RNA or an antisense oligonucleotide inserted into the expression vector and targeting a RBFOX2 coding sequence, wherein the sequence is shown in any one of SEQ ID NO.3 to SEQ ID NO.

5.

6. The recombinant vector targeting RBFOX2 according to claim 5, characterized in that: in, The expression vector is a plasmid vector or a viral vector, and the viral vector is selected from adeno-associated virus or adenovirus.

7. Use of the recombinant vector targeting RBFOX2 according to claim 5 in the preparation of a drug for treating inflammatory bowel disease.

8. A pharmaceutical composition for treating inflammatory bowel disease, characterized in that: The pharmaceutical composition comprises an active ingredient and a medically acceptable excipient, carrier or diluent. Wherein, the active component is the RBFOX2 inhibitor according to any one of claims 1 to 4 or the RBFOX2-targeted recombinant vector according to any one of claims 5 to 6.

9. Use of a reagent for detecting RBFOX2 expression level in the preparation of an inflammatory bowel disease detection kit or a drug screening kit.

10. An inflammatory bowel disease diagnostic kit or drug screening kit, comprising a reagent for detecting the expression level of RBFOX2 in a biological sample.

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