Use of rbfox2 and related substances in the treatment of colitis

Abstract

The present application relates to the technical field of biological medicine engineering, and provides application of RBFOX2 inhibitor in treatment of inflammatory bowel disease, and particularly relates to the role of interfering RNA or antisense oligonucleotide or recombinant expression vector targeting RBFOX2 coding sequence in preparation of products for treating colitis and its related symptoms and / or other complications, and also relates to corresponding recombinant vectors or pharmaceutical compositions. The RBFOX2 inhibitor can effectively reduce the level of proinflammatory cytokines, inhibit the activation of macrophages, has effective application value in relieving and treating colonic inflammation and other symptoms and / or other complications of inflammatory bowel disease, provides a new target for treatment of inflammatory bowel disease, and has wide clinical application prospect.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to RNA binding fox-1 homolog 2 (RBFOX2) as a target in the preparation of drugs for the treatment of inflammatory bowel disease (IBD), and further relates to vectors and pharmaceutical compositions comprising the same. BACKGROUND

[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), the two kinds of IBD have both overlap and difference in clinical and pathological features. In recent years, the incidence and prevalence of IBD in China has shown a significant upward trend. The pathogenesis of IBD is not completely clear at present, and previous studies have shown that the interaction between intestinal microorganisms, intestinal epithelial cells and innate immunity widely participates in the pathogenesis of IBD. Innate immune response plays an important role in it. In the intestinal tract of healthy people, macrophages do not produce pro-inflammatory cytokines, but still have phagocytic activity and antibacterial activity. Studies have shown that in CD patients, CD14 + Macrophages express both macrophage (CD14, CD33 and CD68) and dendritic cell (CD205 and CD209) marker molecules, can produce a large number of inflammatory factors IL-6, IL-23 and TNF, and promote the production of IFN-γ. Although the treatment of IBD has made rapid progress in recent years, it targets a single inflammatory factor or inflammatory pathway, resulting in limited clinical efficacy. Therefore, by studying the pathological process and pathogenic genes of IBD, it is of great practical significance and clinical transformation prospect to fundamentally block and intervene the pathological effects of abnormal activation of immune cells in the inflammatory microenvironment of the colon.

[0003] By analyzing the differentially expressed genes of synovial tissue of IBD patients and healthy people, it is identified that the high expression of RNA binding fox-1 homolog 2 (RBFOX2) in peripheral blood mononuclear cells (PBMC) of IBD patients may be involved in the occurrence and development of IBD. RBFOX2, also known as RBM9 or FOX2, has a full length of 390 amino acids and is highly conserved between yeast and various mammals, indicating that the protein has extremely important biological functions. RBFOX2 is reported to play an important biological role in diabetes, heart failure, fatty liver and pancreatic cancer, myeloid leukemia and other tumors. At present, the research on the protein is less, mainly focusing on the formation of the classic catalytic center of the spliceosome, that is, RBFOX2 can recognize the evolutionarily conserved GCAUG motif to perform alternative splicing on pre-mRNA. It has been found that in the pancreas, RBFOX2 can regulate the alternative splicing of genes required for insulin granule docking and exocytosis, thereby affecting the secretion of insulin; in cardiomyocytes, RBFOX2 can splice multiple key genes in the cGMP-PKG-Ca2 + pathway, causing a significant increase in intracellular calcium levels; in liver cells, RBFOX2 regulates the alternative splicing of class I scavenger receptor Scarb1, maintaining cholesterol homeostasis in the liver lipogenic environment; transforming growth factor TGF-β can induce RBFOX2 to catalyze the alternative splicing of TAK1, playing a role in tumor epithelial-mesenchymal transition and chemotherapy drug resistance. In addition, RBFOX2 can also directly bind to the RNA binding motif 15 (RBM15) of the polycomb complex 2 (PRC2) that mediates H3K27me3 and the N6-methyladenosine methylation (m 6 A) mediated RNA binding motif 15 (RBM15) to affect 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 the high expression of RBFOX2 may be related to the occurrence and development of IBD.

[0004] In summary, there is an urgent need in the art to develop new mitigation and treatment strategies for 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 associated symptoms and / or other complications. SUMMARY

[0005] The present application is directed to the above problem, first confirms the adverse effect of abnormal expression of RBFOX2 on IBD, then observes the recovery of colonic inflammatory damage after the elimination or alleviation of abnormal expression of RBFOX2 based on the target, and further determines the role of RBFOX2 in the onset and progression of IBD.

[0006] The research process of the present application 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 a 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 involved in the IBD disease alleviation process mediated by RBFOX2 deficiency. Silencing the expression of RBFOX2 using specific small interfering RNA (siRNA) targeting the RBFOX2 gene can also down-regulate the production of CXCL1 in mouse peritoneal macrophages. It is concluded that the high expression of RBFOX2 in IBD recruits more immune inflammatory cells by up-regulating the production of CXCL1, and plays an important role in the pathogenesis and progression of IBD.

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

[0009] In a first aspect of the present application, the use of an RBFOX2 inhibitor in the preparation of a medicament for treating IBD or its complications is provided.

[0010] IBD is a macrophage and T lymphocyte pathogenic-induced colonic inflammation. The symptoms related to IBD are colitis-related symptoms characterized by abdominal pain, diarrhea, and bloody stool, and 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 the affected part of the intestine.

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

[0012] The RBFOX2 coding sequence is selected from any of the following:

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

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

[0015] (iii) the RNA sequence of (i) and / or (ii) with one or more nucleotides substituted, deleted or added, and encoding a protein or polypeptide with 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 or having sequence homology (e.g. 80% or more homologous or having 80% or more sequence identity, such as 80%, 85%, 90%, 95%, 98%, 99%) to the amino acid sequence shown in SEQ ID NO. 2, and having the ability to aggravate the symptoms of IBD;

[0019] a protein or polypeptide that aggravates the symptoms of IBD and its related symptoms and / or other complications;

[0020] (c) a protein or polypeptide derived from (a) or (b) with one or several amino acids substituted, deleted or added in the amino acid sequence of (a) or (b), and having the ability to aggravate the symptoms of IBD.

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

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

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

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

[0025] In a second aspect of the present application, there is provided 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 shown above.

[0026] In a third aspect of the present application, there is provided the use of the recombinant vector targeting RBFOX2 in the preparation of a medicament for treating IBD.

[0027] The vector includes a viral vector and a non-viral vector.

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

[0029] The "non-viral vector" includes a liposome or lipid complex, a cationic polymer, a chitosan polymer, and a nanoparticle carrier, etc. Suitable non-viral vectors are well known to those skilled in the art.

[0030] In a fourth aspect, the present application provides a pharmaceutical composition for treating IBD and its complications, comprising an active ingredient and a pharmaceutically acceptable excipient, carrier or diluent. The active ingredient is the RBFOX2 inhibitor or the recombinant vector targeting RBFOX2 as described above.

[0031] The recombinant virus of the present application, together with pharmaceutically acceptable adjuvants, constitutes a pharmaceutical composition for treating IBD, thereby more stably exerting a therapeutic effect.

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

[0033] Generally, the liquid preparation can be stored at 2-8°C for at least one year, and the lyophilized preparation can be stored at 30°C for at least six months. The preparation can be a suspension, an aqueous needle, a lyophilized preparation, etc. commonly used in the pharmaceutical field.

[0034] The dosage of the composition of the present application when administered to animals including humans varies depending on the age and body weight of the patient, the nature and severity of the disease, and the route of administration, and can be determined with reference to the results of animal experiments and various conditions, provided that the total amount of administration does not exceed a certain range.

[0035] Further, the pharmaceutical composition of the present application can be combined with other pharmaceutical compositions for treating IBD-related symptoms or other complications.

[0036] In a fifth aspect, the present application provides the use of a reagent for detecting the expression level of RBFOX2 in the preparation of an IBD detection kit or a drug screening kit.

[0037] The experimental results show that the expression of RBFOX2 is significantly up-regulated in IBD patients, and RBFOX2 knockout alleviates the occurrence and development of DSS-induced colitis in mice, inhibits the activation, infiltration and tissue damage of inflammatory cells in the colon microenvironment, suggesting that RBFOX2 is a promoting factor for IBD and has a promoting effect on the occurrence and development of IBD. Accordingly, it can be inferred that RBFOX2 can be used as a diagnostic marker or a 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 candidate substances in IBD or simulating IBD symptoms or other inflammatory environments;

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

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

[0042] In a sixth aspect of the present application, an IBD diagnostic kit or drug screening kit is provided, which comprises reagents 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. By detecting the nucleic acid expression level of RBFOX2, diagnosis or drug screening is achieved.

[0043] The primer sequence of the primer system is 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'-TCCGAGGAGCCCATCTGAGG-3' (SEQ ID NO. 8);

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

[0048] Advantages and effects of the present application:

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

[0050] Figure 1Disease severity scores and weight loss in DSS-induced RBFOX2 knockout mouse (CKO mouse) colitis model; where A represents disease severity scores for WT and cKO mice; and B represents the weight loss percentage for WT and cKO mice.

[0051] Figure 2 The results show that RBFOX2 knockout reduces the inflammatory response and tissue damage in the colonic microenvironment; A, WT and cKO mouse DSS-induced colitis model results of colonic tissue; B, WT and cKO mouse histopathological scores.

[0052] Figure 3 The results showed that knocking out or interfering with the expression of RBFOX2 reduced the expression level of CXCL1 in mouse peritoneal macrophages; where A represents the mRNA expression level of CXCL1 in mouse peritoneal macrophages with RBFOX2 knockout; B represents the mRNA expression level of CXCL1 in mouse peritoneal macrophages with RBFOX2 interference; and C represents the expression level of CXCL1 in the supernatant of mouse peritoneal macrophages with RBFOX2 knockout.

[0053] Figure 4 This study demonstrates that AAV9 virus interfering with RBFOX2 expression can inhibit CXCL1 expression levels in macrophages and cause pathological damage in colon tissue. A shows the immunoblotting results of RBFOX2 protein expression detected after macrophage infection with recombinant virus AAV9-RBFOX2-RNAi; B shows that recombinant virus AAV9-RBFOX2-RNAi inhibits CXCL1 production in mouse peritoneal macrophages; and C shows the eosin & hematoxylin staining results of colon tissue after tail vein injection of recombinant virus AAV9-RBFOX2-RNAi in IL10- / - mice with spontaneous colitis. Detailed Implementation

[0054] The following examples and experimental cases further illustrate the present invention and should not be construed as limiting the invention. The examples do not include detailed descriptions of conventional methods, such as PCR methods, methods used to construct 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 have been described in numerous 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 indicated, percentages and parts are by volume. As used herein, all technical and scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the application pertains. Furthermore, any method and material similar or equivalent to those described herein can be used in the practice of the present application, and the preferred methods and materials are presented by way of example only.

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

[0057] RBFOX2 myeloid cell conditional knockout mice were constructed. In these mice, RBFOX2 fl / fl Mice were constructed by SinoBiologics Co., Ltd. and then crossed with Lyz2-Cre mice to obtain RBFOX2 fl / fl Lyz2-Cre + / – Mice, referred to as cKO mice, so as to achieve specific knockout of the RBFOX2 gene in myeloid cells, and control mice RBFOX2 fl / fl Lyz2-Cre – / – (referred to as WT mice). These mice were raised in a specific pathogen-free (SPF) environment.

[0058] WT and cKO mice were given 3% DSS drinking water after being fasted for one night to construct a mouse DSS-induced colitis model. The monitoring indicators included body weight change (> 110% 0 points; 90%-110% 1 point; 70%-89% 2 points; <70% 3 points), stool character (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 a total score of up to 9 points.

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

[0060] This result shows that RBFOX2 knockout alleviates the occurrence and development of mouse DSS-induced colitis, 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 Example 1). The colon of the mice was obtained, fixed with paraformaldehyde, and then stained with eosin and hematoxylin to observe the histopathological changes in the colon of the mice.

[0063] The results showed that pathogenic inflammatory cell infiltration was significantly reduced and crypt structure was more intact in the colonic microenvironment of RFBOX2 knockout mice. Figure 2 A). The degree of histopathological damage in the colon tissue of RBFOX2 knockout mice was significantly reduced compared to WT mice, with a lower histological score. Figure 2 B).

[0064] These results indicate that RBFOX2 knockout alleviates inflammatory cell activation, infiltration, and tissue damage in the colonic microenvironment, suggesting the pro-inflammatory role of RBFOX2.

[0065] Example 3: Knocking out or interfering with RFBOX2 expression enhances CXCL1 expression in mouse peritoneal macrophages.

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

[0067] Peritoneal macrophages from wild-type mice were obtained and transfected with siRNA sequences targeting mouse RFBOX2 (sequences shown in SEQ ID NO. 3–5, synthesized by Santa Cruz) and control sequences (both purchased from Santa Cruz). Seventy-two hours after transfection, peritoneal macrophages were stimulated with lipopolysaccharide (LPS) (100 ng / ml) to mimic the activated state of macrophages in IBD. Total RNA from macrophages was collected, and CXCL1 levels were detected by reverse transcription PCR followed by real-time quantitative PCR.

[0068] The specific sequence of the 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 RBFOX2 knockout mice produced lower levels of CXCL1 in their peritoneal macrophages.Figure 3 A) Interference of RBFOX2 in mouse peritoneal macrophages significantly decreased the level of CXCL1 production Figure 3 B) Knocking out RBFOX2 in mouse peritoneal macrophages also significantly decreased the level of CXCL1 in the supernatant Figure 3 C).

[0073] This result shows that reducing the expression of RBFOX2 by using gene knockout or siRNA interference can inhibit the activation of macrophages and reduce the production of CXCL1 in macrophages.

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

[0075] According to the siRNA sequence targeting the RBFOX2 gene in the foregoing Example 3, the Shanghai Jikai Gene Company was commissioned to successfully construct an adeno-associated virus vector based on AAV9 that targets and interferes with the expression of RBFOX2, which was named AAV9-RBFOX2-RNAi. The above three AAV9-RBFOX2-RNAi and control empty virus AAV9-control were used to infect primary peritoneal macrophages, and Western blot was used to detect the infection efficiency of the virus to confirm its interference effect. Then the above viruses were used to infect mouse peritoneal macrophages (same as Example 3), and lipopolysaccharide LPS (100 ng / ml) was used for stimulation to simulate the activation state of macrophages in IBD, and the total RNA of macrophages was collected for reverse transcription PCR and then real-time fluorescence quantitative PCR to detect the level of CXCL1.

[0076] The results show that AAV9-RBFOX2-RNAi( Figure 4 Lane 2, 3, 4) compared with AAV9-control( Figure 4 Lane 1) can be observed that the expression of RBFOX2 protein is significantly reduced; the third AAV9-RBFOX2-RNAi and AAV9-control were used to infect mouse peritoneal macrophages, respectively, and it was found that interfering with the expression of RBFOX2 inhibited the production of CXCL1 in mouse peritoneal macrophages Figure 4 B); the third AAV9-RBFOX2-RNAi and AAV9-control were injected into the tail vein of spontaneous colitis Il10- / - mice, respectively, and the results of eosin & hematoxylin staining of colon tissue showed that interfering with the expression of RBFOX2 significantly reduced the inflammatory damage of colitis Figure 4 C).

[0077] This result shows that targeted inhibition of RBFOX2 can inhibit the activation of macrophages, reduce the production of CXCL1 in macrophages, and reduce the inflammatory response and pathological damage of colitis.

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

[0079]

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

[0081] MEKKKMVTQGNQEPTTTPDAMVQPFTTIPPFPPPPQNGIPTEYGVPHTQDYAGQTGEHNLTLYGSTQAHGEQSSNSPSTQNGSLTTEGGAQTDGQQSQTQSSENSESKSTPKRLHVSNIPFRFRDPDLRQMFGQFGKILDVEIIFNERGSKGFGFVTFENSADADRAREKLHGTVVEGRKIEVNNATARVM TNKKMVTPYANGWKLSPVVGAVYGPELYAASSFQADVSLGNDAAVPLSGRGGINTYIPLISLPLVPGFPYPTAATTAAAFRGAHLRGRGRTVYGAVRAVPPTAIPAYPGVVYQDGFYGADLYGGYAAYRYAQPATATAATAAAAAAAAYSDGYGRVYTADPYHALAPAASYGVGAVASLYRGGYSRFAPY

[0082] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. The use of RPFOX2 inhibitors in the preparation of drugs for treating inflammatory bowel disease, characterized in that, The inflammatory bowel disease is ulcerative colitis; the RBFOX2 inhibitor is an interfering RNA or antisense oligonucleotide targeting the RBFOX2 coding sequence, the nucleotide sequence of which is shown in SEQ ID NO.

5.

2. A recombinant vector targeting RFBOX2, characterized in that, It includes an expression vector and an interfering RNA or antisense oligonucleotide inserted into the expression vector that targets the RBFOX2 coding sequence, as shown in SEQ ID NO.

5.

3. The recombinant vector targeting RFBOX2 according to claim 2, 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.

4. The use of the recombinant vector targeting RPFOX2 according to claim 2 in the preparation of a drug for treating inflammatory bowel disease, characterized in that, The inflammatory bowel disease mentioned is ulcerative colitis.

5. A pharmaceutical composition for treating ulcerative colitis, characterized in that, The pharmaceutical composition includes an active ingredient and medically acceptable excipients, carriers, or diluents. The active component is the RBFOX2 inhibitor of claim 1 or the RBFOX2-targeting recombinant vector of claim 2.

6. The application of reagents for detecting RFBOX2 expression levels in the preparation of inflammatory bowel disease detection kits or inflammatory bowel disease drug screening kits, characterized in that, The inflammatory bowel disease mentioned is ulcerative colitis.

Images