Composition for diagnosis and treatment of inflammatory bowel disease comprising mirna as active ingredient

Abstract

The present invention provides a miRNA biomarker for diagnosing and treating inflammatory bowel disease (IBD). A biomarker according to one embodiment of the present invention can be utilized in a composition for diagnosing inflammatory bowel disease, a method for providing information for diagnosis, a pharmaceutical composition for preventing or treating inflammatory bowel disease, and a method for treating inflammatory bowel disease.

Description

Composition for diagnosis and treatment of inflammatory bowel disease containing MIRNA as an active ingredient

[0001] The present invention provides a miRNA biomarker for the diagnosis and treatment of inflammatory bowel disease (IBD) by isolating terminal ileum tissue from patients with IBD and normal controls and then selecting miRNAs that can be utilized as biomarkers through small RNA sequencing. This biomarker can be utilized in a composition for diagnosing inflammatory bowel disease, a method for providing information for diagnosis, a pharmaceutical composition for preventing or treating inflammatory bowel disease, and a method for treating inflammatory bowel disease.

[0002] Inflammatory bowel disease is a chronic inflammatory disease that causes inflammation or ulcers in the intestines for an unknown cause. It is a chronic, intractable disease that is broadly divided into Crohn's disease, ulcerative colitis, and intestinal Behcet's disease.

[0003] Crohn's disease (CD) is a chronic, intractable inflammatory bowel disease (IBD) that can affect any part of the gastrointestinal tract, from the mouth to the anus. The primary symptoms of CD include abdominal pain, diarrhea, and weight loss. It typically develops in young adults, typically in their teens or twenties, persists throughout life, and can lead to complications such as intestinal strictures, fistulas, and perforations. The pathogenesis of CD is believed to be complex, involving genetic and environmental factors, as well as an abnormal immune response to intestinal flora. However, its exact cause remains unclear.

[0004] Ulcerative colitis is a chronic inflammatory bowel disease of unknown etiology characterized by inflammation confined to the mucosa or submucosa of the colon. Its main symptoms include bloody diarrhea with recurrent flare-ups, urgency, and abdominal pain. Ulcerative colitis is caused by the interaction of genetic susceptibility, intestinal microflora, external stimuli, and the body's immune system.

[0005] Inflammatory bowel disease (IBD) is a common condition primarily in developed Western countries, but has been extremely rare in East Asian countries, including Korea. However, recent epidemiological studies show a steady increase in the incidence of IBD in East Asia, including Korea. In Korea, the incidence has increased approximately 2.32-fold over the past 10 years (2020 IBD Factsheet, Korean Society for the Study of the Intestine).

[0006] Inflammatory bowel disease cannot be accurately diagnosed with just one method, and is currently diagnosed by synthesizing symptoms, endoscopic and tissue examinations, blood tests, and imaging examination findings.

[0007] Furthermore, various treatment strategies have been attempted for inflammatory bowel disease, targeting diverse clinical manifestations and complications. Recently, treatment strategies based on numerous clinical research findings have been proposed. Pharmacological treatment is primarily administered, but surgical intervention is indicated in cases of complications such as intestinal obstruction, stricture, or perforation. Medications include anti-inflammatory drugs, steroids (adrenal corticosteroids), immunomodulators, and biological agents.

[0008] Since the 1980s, thiopurine agents, including the immunomodulators azathioprine and 6-mercaptopurine, have been widely used to maintain remission in steroid-dependent or refractory Crohn's disease and ulcerative colitis, and to prevent postoperative relapse. However, potential side effects, such as bone marrow suppression, hepatotoxicity, opportunistic infections, and the risk of malignancy, necessitate the development of treatments for inflammatory bowel disease with fewer side effects.

[0009] In addition, biological agents that have been used since 1998 are very expensive drugs that can cause side effects such as pulmonary tuberculosis, allergic reactions, skin diseases, and malignant tumors. In addition, regardless of the type of drug, approximately 20-30% of patients do not respond from the beginning of use, and even among patients who initially respond, approximately 20-30% lose their drug response within one year of treatment. Therefore, there is a high demand for drugs with new mechanisms of action.

[0010] In order to solve the above problems, the inventors of the present invention isolated terminal ileum tissues of inflammatory bowel disease and compared the differences in miRNA expression between inflammatory bowel disease patients and normal controls, thereby selecting miRNAs that can be used as biomarkers for effectively diagnosing and treating inflammatory bowel disease, and developing and providing pharmaceutical compositions and food compositions for diagnosing or treating inflammatory bowel disease that include the miRNAs as active ingredients.

[0011] In addition, the present invention provides a method for diagnosing inflammatory bowel disease using the above biomarker, a method for treating inflammatory bowel disease using the above pharmaceutical composition, and a method for screening an inflammatory bowel disease treatment agent using the above biomarker.

[0012] To achieve the above purpose, the present invention provides a composition for diagnosing inflammatory bowel disease (IBD) that is effective and has few side effects by using a miRNA biomarker.

[0013] In addition, the present invention provides a method for diagnosing inflammatory bowel disease using the miRNA biomarker.

[0014] In addition, the present invention provides a kit capable of diagnosing inflammatory bowel disease by measuring and comparing the miRNA biomarker from a subject.

[0015] In addition, the present invention provides a pharmaceutical composition for preventing or treating inflammatory bowel disease, which comprises the miRNA as an active ingredient.

[0016] In addition, the present invention provides a food composition for preventing or improving inflammatory bowel disease, which contains the miRNA as an active ingredient.

[0017] Additionally, the present invention provides a method for treating inflammatory bowel disease using the miRNA biomarker.

[0018] In addition, the present invention provides a method for screening an inflammatory bowel disease treatment agent through measurement and comparison of the above miRNA.

[0019]

[0020] The terminology used herein is for the purpose of description only and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, the terms "comprises" or "has" and the like are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0021] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0022]

[0023] Hereinafter, the present invention will be described in more detail.

[0024] The miRNA of the present invention may be a miRNA (hsa-miRNA) isolated from human intestinal tissue or a mimic miRNA prepared with an equivalent sequence thereto.

[0025] The miRNAs of the present invention include: miR-31-5p, MiR-192-5p, MiR-215-5p, MiR-194-5p, MiR-338-5p, MiR-3065-5p, MiR-3195, MiR-378a-3p, MiR-192-3p, MiR-125b-5p, MiR-200b-3p, MiR-424-5p, miR-215-3p,miR-503-5p,miR-141-3p,miR-543,miR-194-3p,miR-29b-3p,miR-338-3p,miR-223-3p,miR-548h-5p,miR-548aa,miR-4684-5p, miR-585-3p,miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704 miR-1299,miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p, It may be at least one selected from the group consisting of miR-142-5p, miR-155-5p, miR-6820-3p, and miR-552-3p.

[0026] The above miRNA is not limited, but may preferably be miR-338-3p or miR-378a-3p.

[0027] Inflammatory bowel disease in the present invention includes Crohn's disease, ulcerative colitis, intestinal Behcet's disease, hemorrhagic rectal ulcer, ileal pouchitis or intestinal lesions accompanying therewith.

[0028] The histological score (Histologic Colitis Scoring Method) in the present invention can be evaluated by four scales: severity of inflammation, extent of inflammation, degree of crypt damage, and overall extent of impact.

[0029] The severity of inflammation is scored on a scale from 0 to 3, depending on the intensity of the inflammatory response, which can be evaluated as no inflammation at all (0 points), mild inflammatory response (1 point), moderate inflammatory response (2 points), and severe inflammatory response (3 points), respectively.

[0030] The extent of inflammation is assessed based on the extent to which inflammation is distributed within the intestinal mucosa, and can be scored as no inflammation (0 points), limited to the mucosa (1 point), spanning the mucosa and submucosa (2 points), or inflammation penetrating the entire intestinal wall (3 points).

[0031] The degree of crypt damage is assessed by assessing the level of damage to the crypt structure, which is the digestive tract of the intestinal wall, and can be evaluated as follows: no crypt damage at all (0 points), damage to 1 / 3 of the crypt base (1 point), damage to 2 / 3 of the crypt base (2 points), complete loss of the crypt with only the surface epithelium remaining (3 points), and loss of both the crypt and the surface epithelium (4 points).

[0032] The overall extent of the impact is scored from 0 to 4 points, depending on the proportion of inflammation or damage in the gut, which can be evaluated as 0% (0 points), 1%-25% (1 point), 26%-50% (2 points), 51%-75% (3 points), and 76%-100% (4 points), respectively.

[0033] In the present invention, the disease activity index score (DAI score) can be evaluated by three scales: weight loss, stool consistency, and the presence or absence of blood in the stool.

[0034] Weight loss is assessed on a scale from 0 to 3 based on the percentage of weight lost, which can be rated as no weight loss (0 points), 1-5% weight loss (1 point), 5-10% weight loss (2 points), and 10-20% weight loss (3 points).

[0035] Stool consistency refers to how hard or loose the stool is and can be scored as normal stool (0 points), loose stool (1 point), watery diarrhea (2 points), diarrhea with a little blood (3 points), or watery diarrhea mixed with blood (4 points).

[0036] The presence of blood in the stool can be assessed by assessing whether blood is visible in the stool, with a score of 0 if no blood is visible at all or 4 if blood is visible.

[0037] The pharmaceutical composition according to the present invention may additionally include, in addition to the active ingredient miRNA, a pharmaceutically acceptable carrier, binder, lubricant, disintegrant, excipient, solubilizer, dispersant, stabilizer, suspending agent, etc., and examples thereof may include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, alginate, calcium phosphate, calcium silicate, maltodextrin, silicon dioxide, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, etc.

[0038] In addition, the pharmaceutical composition can be prepared in the form of tablets, capsules, sustained-release preparations, elixirs, suspensions, syrups, wafers, unit dose injection ampoules, suspensions, etc. by methods commonly used in formulation in the art.

[0039] The above pharmaceutical composition may be administered by oral administration, intravenous administration, intramuscular administration, intraarterial administration, intramedullary administration, intrathecal administration, intracardiac administration, transdermal administration, subcutaneous administration, intraperitoneal administration, intranasal administration, enteral administration, local administration including oral and sublingual administration, rectal administration, topical administration, dermal administration, mucosal administration, intraspinal administration, etc., and is preferably administered orally, but is not limited thereto.

[0040] In addition, the above-mentioned administration may be administered by being formulated into a dosage form appropriate for each administration method, and the formulation may be formulated by a method widely known in the art.

[0041] The above pharmaceutical composition may be appropriately selected based on various factors including the age, weight, health condition, sex, administration time, administration route, excretion rate, drug combination, and severity of a specific disease to be prevented or treated of the subject of administration, and the frequency of administration may be once, twice, or three times a day, every other day, every other week, every other month, etc.

[0042] The above food composition can be manufactured in the form of a beverage, gum, tea, vitamin complex, powder, granules, tablets, capsules, confectionery, etc.

[0043] In addition, the food composition may additionally include, in addition to the active ingredient miRNA, flavoring agents, stabilizers, thickeners, preservatives, etc., commonly used in the art to manufacture foods.

[0044] The present invention can provide a miRNA biomarker specific for inflammatory bowel disease, and can provide a composition for diagnosing inflammatory bowel disease comprising the same as an active ingredient. In addition, the miRNA of the present invention has the effect of alleviating symptoms of inflammatory bowel disease such as weight gain and increased intestinal length, reducing inflammatory response mediating cells, reducing inflammatory cell infiltration and subcutaneous tissue edema, and reducing immune cell infiltration, thereby providing a pharmaceutical composition having an excellent effect in preventing or treating inflammatory bowel disease and a food composition having an excellent effect in preventing or improving inflammatory bowel disease.

[0045] Figure 1 shows the overall process and effect of an embodiment and experimental example of the present invention.

[0046] Figure 2 shows the 52 miRNAs that were initially selected through small RNA sequencing after extraction from the sample and showed an expression difference of more than 2-fold between each group.

[0047] Figure 3 shows the expression levels in each tissue sample by performing RT-PCR on 8 miRNAs with the largest difference between groups among the 52 miRNAs initially selected.

[0048] Figure 4 illustrates the process of establishing an inflammatory bowel disease mouse model to confirm the therapeutic efficacy of the final selected miRNAs, miR-338-3p and miR-378a-3p, on inflammatory bowel disease.

[0049] Figure 5 shows that when miR-338-3p, miR-378a-3p, and mixture were injected into an inflammatory bowel disease mouse model, body weight increased compared to the control group.

[0050] Figure 6 shows that when miR-338-3p, miR-378a-3p, and mixture were injected into an inflammatory bowel disease mouse model, the disease activity index decreased and the intestinal length increased compared to the control group.

[0051] Figure 7 shows that when miR-338-3p, miR-378a-3p, and mixture were injected into an inflammatory bowel disease mouse model, the number of Th17 cells, which are the main cells mediating the inflammatory response in the spleen, mesenteric lymph node (MLN), and colon, decreased.

[0052] Figure 8 shows that when miR-338-3p, miR-378a-3p, and mixture were injected into an inflammatory bowel disease mouse model, the histological score, which was evaluated by the degree of inflammatory cell infiltration and subcutaneous tissue edema, was significantly lower than that of the control group.

[0053] Figure 9 shows that when miR-338-3p mimic, miR-378a-3p mimic, and mixture were injected into an inflammatory bowel disease mouse model, the infiltration of macrophages and T cells was significantly reduced compared to the control group.

[0054] Figure 10 shows that when miR-338-3p mimic, miR-378a-3p mimic, and mixture were injected into an inflammatory bowel disease mouse model, IL-33 expression was reduced compared to the control group.

[0055] Figure 11 shows that when miR-338-3p mimic, miR-378a-3p mimic, and mixture were injected into an inflammatory bowel disease mouse model, MACC1 expression was reduced compared to the control group.

[0056] Figure 12 shows the changes in the expression of MACC1 mRNA (Figure 12A) and protein (Figure 12B) according to IL-33 treatment in HT-29 cell lines, and the correlation between IL-33 and MACC1 expression through Pearson correlation analysis (Figure 12C).

[0057] The present invention is described in detail below through examples and experimental examples. The objectives, features, and advantages of the present invention will be readily understood through the following examples. The present invention is not limited to the examples described herein and may be embodied in other forms. The examples introduced herein are provided to ensure that those skilled in the art can sufficiently convey the spirit of the present invention. Therefore, the present invention should not be limited by the following examples.

[0058]

[0059] Prior to the examples, the overall process and effects of the examples and experimental examples of the present invention are shown in Figure 1.

[0060] Example 1: Primary screening of miRNAs showing differences between normal controls and patients with inflammatory bowel disease.

[0061] Terminal ileum tissues were obtained through ileocolonoscopy from normal controls, patients with Crohn's disease in remission, non-inflamed areas of patients with active Crohn's disease, and inflamed areas of patients with active Crohn's disease. For remission, patients with a Crohn's Disease Activity Index (CDAI) of less than 150 and endoscopically healed mucosa were considered to be in remission, and terminal ileum tissues were obtained.

[0062] RNA was extracted from the sample, and a total of 52 miRNAs (hsa-miR-31-5p, hsa-miR-192-5p, hsa-miR-215-5p, hsa-miR-194-5p, hsa-miR-338-5p, hsa-miR-3065-5p, hsa-miR-3195, hsa-miR-378a-3p, hsa-miR-192-3p, hsa-miR-125b-5p, hsa-miR-200b-3p, hsa-miR-424-5p, hsa-miR-215-3p, hsa-miR-503-5p, hsa-miR-141-3p, hsa-miR-543, hsa-miR-194-3p, hsa-miR-29b-3p, hsa-miR-338-3p, hsa-miR-223-3p, hsa-miR-548h-5p, hsa-miR-548aa, hsa-miR-4684-5p, hsa-miR-585-3p, hsa-miR-518b, hsa-miR-552-5p, hsa-miR-4634, hsa-miR-183-3p, hsa-miR-146a-3p, hsa-miR-4485-3p, hsa-miR-20a-5p, hsa-miR-185-5p, hsa-miR-708-5p, hsa-miR-4497, hsa-miR-4792, hsa-miR-7641, hsa-miR-7704, hsa-miR-1299, hsa-miR-125b-1-3p, hsa-miR-424-3p, hsa-miR-5096, hsa-miR-5787, hsa-miR-1273g-3p, hsa-miR-6836-3p, hsa-miR-1972, hsa-miR-202-5p, hsa-miR-509-3-5p, hsa-miR-508-3p, hsa-miR-142-5p, hsa-miR-155-5p, hsa-miR-6820-3p, hsa-miR-552-3p) were selected in the first stage and are shown in Fig. 2.

[0063]

[0064] Example 2: Secondary selection of 8 miRNAs from the first-selected miRNAs

[0065] Among a total of 52 miRNAs, 8 miRNAs (hsa-miR-338-3p, hsa-miR-29b-3p, hsa-miR-141-3p, hsa-miR-125b-5p, hsa-miR-378a-3p, hsa-miR-150-5p, hsa-miR-155p-5p, hsa-miR-200b-3p) with high expression levels and the greatest differences between groups were selected in a second round (cut-off value: normalized data > 8-9, P < 0.05), and RT-PCR was performed on these 8 miRNAs to confirm the expression levels in each tissue sample, which are shown in Fig. 3.

[0066]

[0067] Example 3: Final selection of biomarker miRNAs from the secondary selected miRNAs.

[0068] Eight selected miRNAs were identified in each sample, and hsa-miR-338-3p, hsa-miR-125b-5p, and hsa-miR-378a-3p, which had not been studied previously, were found to have significant differences in expression levels between the normal group and Crohn's disease patients. Based on the PCR results, hsa-miR-338-3p and hsa-miR-378a-3p were selected as miRNAs that could be used as diagnostic biomarkers for inflammatory bowel disease.

[0069] Through the above results, it was confirmed that the final selected miRNAs, hsa-miR-338-3p and hsa-miR-378a-3p, can be used as effective biomarkers for diagnosing inflammatory bowel disease.

[0070]

[0071] Hereinafter, the present invention will be described through experimental examples. However, the scope of the present invention is not limited to the following experimental examples.

[0072] Experimental Example 1: Induction of inflammatory bowel disease and miRNA injection in mice.

[0073] To represent the Crohn's disease patient group, a mouse model of inflammatory bowel disease was established by administering 2.5% dextran sulfate sodium (DSS) to mice for 5 days. The final miRNA mimics (miR-338-3p, miR-378a-3p) were manufactured and ordered, and administered once a week before DSS administration, and again simultaneously with DSS. The miR-338-3p mimic, miR-378a-3p mimic, and miR-mixture (simultaneous administration of miR-338-3p and miR-378a-3p) groups were used as test groups to test the therapeutic efficacy. The process of establishing the inflammatory bowel disease mouse model is illustrated in Figure 4.

[0074]

[0075] Experimental Example 2: Confirming the therapeutic effect of selected miRNAs on inflammatory bowel disease symptoms.

[0076] Afterwards, miR-338-3p mimic, miR-378a-3p mimic, and miR-mixture were administered to the inflammatory bowel disease mouse model to establish an experimental group, and a control group was established by administering a random RNA sequence that does not affect gene expression.

[0077] When miR-338-3p mimic, miR-378a-3p mimic, and miR-mixture were administered to a mouse model of inflammatory bowel disease, body weight increased, DAI score decreased, and intestinal length significantly increased compared to the control group. The effects of selected miRNAs on body weight increase, DAI score decrease, and intestinal length increase are shown in Figures 5 and 6, respectively.

[0078]

[0079] Experimental Example 3: Confirmation of the inflammatory response-mediated cell reduction effect of selected miRNAs.

[0080] When miR-338-3p mimic, miR-378a-3p mimic, and miR-mixture were administered to a mouse model of inflammatory bowel disease, Th17 cells, the main cells mediating the inflammatory response in the spleen and mesenteric lymph nodes (MLN), were significantly reduced compared to the control group. In the colon, Th17 cells were confirmed to be reduced in all groups administered miR-338-3p mimic, miR-378a-3p mimic, and mixture. These results are shown in Fig. 7.

[0081]

[0082] Experimental Example 4: Confirmation of the effect of selected miRNAs on reducing inflammatory cell infiltration and subcutaneous tissue edema.

[0083] When the intestinal tissue of an inflammatory bowel disease mouse model was stained with H&E staining and divided into the proximal, middle, and distal parts and the histological score was evaluated based on the degree of inflammatory cell infiltration and sub-tissue edema, it was confirmed that the histological scores in the miR-338-3p mimic, miR-378a-3p mimic, and mixture administration groups were all significantly lower than those in the control group. These results are shown in Fig. 8.

[0084]

[0085] Experimental Example 5: Confirmation of the effect of selected miRNAs on reducing immune-related cell infiltration.

[0086] The infiltrated macrophages and T cells in a mouse model of inflammatory bowel disease were quantified by immunohistochemistry (IHC). The results showed that, compared to the control group, the infiltration of macrophages and T cells was significantly reduced in the miR-338-3p mimic, miR-378a-3p mimic, and mixture administration groups. These results are shown in Figure 9.

[0087] Experimental Example 6: Confirmation of the effect of selected miRNAs on reducing IL-33 and MACC1 mRNA expression.

[0088] The mRNA targets of miR-338-3p and miR-378a-3p were predicted through analysis using two miRNA target prediction programs (TargetScan, DIANA microT), and MACC1 and IL-33 were selected as mRNA target candidates of miR-338-3p and miR-378a-3p, respectively, through literature review.

[0089] Accordingly, IL-33 and MACC1 expression was analyzed by IHC in the intestinal tissues of mice administered miR-338-3p mimic, miR-378a-3p mimic, and mixture. As a result, it was confirmed that MACC1 expression was reduced in the miR-338-3p administration group, and IL-33 and MACC1 expression was reduced in the miR-378a-3p mimic and mixture administration groups. The IL-33 expression reduction effect of the selected miRNA is shown in Fig. 10, and the MACC1 expression reduction effect is shown in Fig. 11.

[0090]

[0091] Experimental Example 7: Identification of the Cause of MACC1 Decreased Following Administration of miR-378a-3p Through Correlation Analysis Between IL-33 and MACC1

[0092] To determine the cause of the decrease in MACC1 expression by miR-378a-3p administration, HT-29 cells, a colon cancer cell line, were treated with 10 ng / mL IL-33, and it was confirmed that MACC1 expression increased at both mRNA and protein levels.

[0093] In addition, Pearson correlation analysis was performed using RNA seq datasets (GSE102746, GSE117875, GSE66207, GSE83687, GSE135223) of 98 control subjects and 120 IBD patients, and a positive correlation was confirmed between IL-33 and MACC1 with r=0.417 (p<0.001).

[0094] Through this, it was confirmed that IL-33 expression decreased following administration of miR-378a-3p mimic, and that MACC1 expression decreased due to the decreased IL-33 expression. These results are shown in Fig. 12.

[0095]

[0096] Through the above results, it was confirmed that the final selected miRNAs, miR-338-3p and miR-378a-3p, can act as effective ingredients in the treatment of inflammatory bowel disease.

[0097]

[0098] As a result of the above, it is possible to effectively diagnose inflammatory bowel disease using the miRNA of the present invention, and the treatment effects of inflammatory bowel disease, such as weight gain, intestinal length increase, and inflammation reduction, can be clearly confirmed through mouse inoculation experiments, so it is judged that inflammatory bowel disease can be effectively treated.

[0099]

[0100] Although the preferred embodiments and experimental examples of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements that can be made by a person skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0101] The present invention can provide a miRNA biomarker specific for inflammatory bowel disease, and can provide a composition for diagnosing inflammatory bowel disease comprising the same as an active ingredient. In addition, the miRNA of the present invention has the effect of alleviating symptoms of inflammatory bowel disease such as weight gain and increased intestinal length, reducing inflammatory response mediating cells, reducing inflammatory cell infiltration and subcutaneous tissue edema, and reducing immune cell infiltration, and thus can provide a pharmaceutical composition having an excellent effect in preventing or treating inflammatory bowel disease and a food composition having an excellent effect in preventing or improving inflammatory bowel disease, and thus has industrial applicability.

Claims

1. A preparation comprising a preparation for measuring the level of a miRNA biomarker associated with inflammatory bowel disease, The miRNA biomarkers include miR-31-5p, miR-192-5p, miR-215-5p, miR-194-5p, miR-338-5p, miR-3065-5p, miR-3195, miR-378a-3p, miR-192-3p, miR-125b-5p, miR-200b-3p, miR-424-5p, miR-215-3p,miR-503-5p,miR-141-3p,miR-543,miR-194-3p,miR-29b-3p,miR-338-3p,miR-223-3p,miR-548h-5p,miR-548aa,miR-4684-5p, miR-585-3p,miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704 miR-1299,miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p, A composition for diagnosing inflammatory bowel disease, characterized in that it comprises at least one selected from the group consisting of miR-142-5p, miR-155-5p, miR-6820-3p, and miR-552-3p.

2. In claim 1, A composition for diagnosing inflammatory bowel disease, characterized in that the above miRNA biomarker is miR-338-3p or miR-378a-3p.

3. In claim 1 or claim 2, A composition for diagnosing inflammatory bowel disease, characterized in that the inflammatory bowel disease is at least one selected from the group consisting of Crohn's disease, ulcerative colitis, intestinal Behcet's disease, hemorrhagic rectal ulcer, ileal pouchitis, and intestinal lesions accompanying the same. 4.(a) A step of isolating terminal ileum tissue from the subject; (b) a step of measuring the expression level of miRNA biomarkers from the separated terminal ileum tissue; and (c) a step of comparing the measured miRNA expression level with the miRNA biomarker expression level of the normal control group; (d) a step of determining the subject as having inflammatory bowel disease when the expression level of the miRNA biomarker measured in the above step (c) is decreased compared to the control group; including, The miRNA biomarkers include miR-31-5p, miR-192-5p, miR-215-5p, miR-194-5p, miR-338-5p, miR-3065-5p, miR-3195, miR-378a-3p, miR-192-3p, miR-125b-5p, miR-200b-3p, miR-424-5p, miR-215-3p,miR-503-5p,miR-141-3p,miR-543,miR-194-3p,miR-29b-3p,miR-338-3p,miR-223-3p,miR-548h-5p,miR-548aa,miR-4684-5p, miR-585-3p,miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704 miR-1299,miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p, A method for diagnosing inflammatory bowel disease, characterized in that at least one selected from the group consisting of miR-142-5p, miR-155-5p, miR-6820-3p, and miR-552-3p.

5. In claim 4, A method for diagnosing inflammatory bowel disease, wherein the above miRNA biomarker is miR-338-3p or miR-378a-3p.

6. A kit for diagnosing inflammatory bowel disease, comprising the composition of paragraph 1 or 2 as an active ingredient.

7. In claim 6, A kit for diagnosing inflammatory bowel disease, characterized in that the kit further includes a description of a method for diagnosing inflammatory bowel disease comprising the following steps: (a) a step of measuring the expression level of miR-338-3p or miR-378a-3p in terminal ileum tissue isolated from a subject; (b) a step of comparing the expression level of miR-338-3p or miR-378a-3p measured in the above step (a) with the control group; and (c) A step of determining that the subject has inflammatory bowel disease when the expression level of miR-338-3p or miR-378a-3p measured in the above step (b) is decreased compared to the control group. 8.miR-31-5p,miR-192-5p,miR-215-5p,miR-194-5p,miR-338-5p,miR-3065-5p,miR-3195,miR-378a-3p,miR-192-3p,miR-125b-5p,miR-200b-3p,miR-424-5p, miR-215-3p,miR-503-5p,miR-141-3p,mimi543,mir-194-3p,mir-29b-3p,mir-338-3p,mir-223-3p,mir-548h-5p,mir-548aa,mir-4684-5p,mir-585-3p, miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704,miR-1299 miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p,mimi-142-5p, A pharmaceutical composition for preventing or treating inflammatory bowel disease, comprising as an active ingredient at least one miRNA selected from the group consisting of miR-155-5p, miR-6820-3p, and miR-552-3p.

9. In claim 8, A pharmaceutical composition for treating inflammatory bowel disease, characterized in that the above miRNA is miR-338-3p or miR-378a-3p.

10. In claim 8 or claim 9, A pharmaceutical composition for preventing or treating inflammatory bowel disease, characterized in that the inflammatory bowel disease is at least one selected from the group consisting of Crohn's disease, ulcerative colitis, intestinal Behcet's disease, hemorrhagic rectal ulcer, ileal pouchitis and intestinal lesions accompanying the same. 11.miR-31-5p; miR-215-3p,miR-503-5p,miR-141-3p,mimi543,mir-194-3p,mir-29b-3p,mir-338-3p,mir-223-3p,mir-548h-5p,mir-548aa,mir-4684-5p,mir-585-3p, miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704,miR-1299 miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p,mimi-142-5p, A food composition for preventing or improving inflammatory bowel disease, comprising as an active ingredient at least one miRNA selected from the group consisting of miR-155-5p, miR-6820-3p, and miR-552-3p.

12. In claim 11, A food composition for preventing or improving inflammatory bowel disease, characterized in that the above miRNA is miR-338-3p or miR-378a-3p.

13. In claim 11 or claim 12, A food composition for preventing or improving inflammatory bowel disease, characterized in that the inflammatory bowel disease is at least one selected from the group consisting of Crohn's disease, ulcerative colitis, intestinal Behcet's disease, hemorrhagic rectal ulcer, ileal pouchitis and intestinal lesions accompanying the same.

14. A method for preventing or treating inflammatory bowel disease, comprising administering the pharmaceutical composition of claim 8 or claim 9. 15.(a) A step of treating the target substance to be screened in cells, tissues or non-human animals separated as an experimental group; and (b) a step of measuring and comparing the expression level of miRNA in the experimental group of step (a) and the control group that was not treated with the target substance; a screening method for a treatment agent for inflammatory bowel disease, comprising: The above-mentioned miRNAs include: miR-31-5p, miR-192-5p, miR-215-5p, miR-194-5p, miR-338-5p, miR-3065-5p, miR-3195, miR-378a-3p, miR-192-3p, miR-125b-5p, miR-200b-3p, miR-424-5p, miR-215-3p,miR-503-5p,miR-141-3p,mimi543,mir-194-3p,mir-29b-3p,mir-338-3p,mir-223-3p,mir-548h-5p,mir-548aa,mir-4684-5p,mir-585-3p, miR-518b,miR-552-5p,miR-4634,miR-183-3p,miR-146a-3p,miR-4485-3p,miR-20a-5p,miR-185-5p,miR-708-5p,miR-4497,miR-4792,miR-7641,miR-7704,miR-1299 miR-125b-1-3p,miR-424-3p,miR-5096,miR-5787,miR-1273g-3p,miR-6836-3p,miR-1972,miR-202-5p,miR-509-3-5p,miR-508-3p,mimi-142-5p, A screening method, wherein at least one selected from the group consisting of miR-155-5p, miR-6820-3p, and miR-552-3p.

16. In paragraph 15, A method for screening an inflammatory bowel disease treatment agent, wherein the miRNA is miR-338-3p or miR-378a-3p.

17. In paragraph 15 or 16, A method for screening an inflammatory bowel disease treatment agent, characterized in that the inflammatory bowel disease is at least one selected from the group consisting of Crohn's disease, ulcerative colitis, intestinal Behcet's disease, hemorrhagic rectal ulcer, ileal pouchitis, and intestinal lesions accompanying the same.

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