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Role of Host—Microbial Interactions in Inflammatory Bowel Disease (IBD)
Japan (IFMSA-Japan) - Kurume University, Kurume
Department of Immunology
Type of Research Project
- Basic science
What is the background of the project?
In 2005, our group identified Chitinase 3-like 1 (CHI3L1 or YKL-40), as an intestinal inflammation associated inducible molecule, which enhances both bacterial adhesion and invasion on/into intestinal epithelial cells (IECs). Based on our published and preliminary data, we plan to prove that inflammation-induced CHI3L1 expression exacerbates chronic intestinal inflammation by enhancing the interaction of IECs with potentially pathogenic bacteria permanently and subsequently induce colitis-associated neoplastic changes in IECs.
What is the aim of the project?
The aim of this project is to define how an epithelial cell-derived molecule termed CHI3L1 interacts with potentially pathogenic bacteria in normal microflora during the development of inflammatory bowel disease(IBD)and colitis-associated cancer.In addition,we will test the therapeutic effects of methylxanthine derivatives including caffeine and theophylline, which are targeting pan-chitinases during the development of IBD using both in vitro and in vivo systems in our laboratory.
What techniques and methods are used?
General theoretical teaching will be supervised/instructed by the tutor as well as our lab members. For example, the student will extract Ribonucleic acid(RNA) from HCT116 human colon cancer cells with RNA-zol method followed by complementary DNA (cDNA) synthesis using Invitrogen SuperScripts II kit. CHI3L1 as well as β-actin mRNA expression levels will be analyzed by Reverse transcription polymerase chain reaction(RT-PCR) and Quantitative polymerase chain reaction(Q-PCR). Protein samples will be prepared from HCT116 and/or animals (C57BL/6 wild-type mice and CHI3L1 knockout mice) extracting from lymphoid organs (spleen, mesenteric lymph nodes, thymus, colon, and small intestine) to check the baseline expression of CHI3L1, acidic mammalian chitinase (AMCase) and chitinase-1 by immunohistochemistry and Western blot analysis. We will also analyze the expression of these proteins after inducing acute colitis by orally administrating 3.5% dextran sulfate sodium (DSS) in the same strains of mice. Ribosomal 16S Deoxyribonucleic acid (DNA) will be extracted from fecal contents and intestinal mucosa associated bacterial samples, and the bacterial compositions from these samples will be analyzed before and after inducing DSS colitis.Severity of colitis / bacterial composition change will be analyzed treating these mice with chitinase inhibitors (e.g. caffeine, theophylline) and/or anti-chitinase antibodies.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
What are the tasks expected to be accomplished by the student?
_The student will help lab technicians, undergraduate/graduate students, post-doctoral fellows and assistant/associate professors in preparing instructions and materials used during laboratory experiments, which includes cancer cell culture, flow cytometry analysis, immunohistochemistry, Western blot analysis, animal dissections, and some basic molecular biology techniques, including DNA/RNA extraction, cDNA synthesis, agarose gel electrophoresis, primer design, RT-PCR (reverse transcription polymerase chain reaction) and Q-PCR (quantitative real-time PCR) analyses. The students will perform a couple of experiments under the supervision of lab members. He/she needs to complete the online lab safety work in the biology lab before starting to work.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a presentation - The student will prepare a scientific report - The student will prepare an abstract - The student’s name will be mentioned in a future publication
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic knowledge on immunology/cellular biology as well as basic lab skills,including basic steps in sterile techniques and animal handling,will be absolutely necessary before joining the lab.The students need to design and perform experiments semi-independently.
Are there any legal limitations in the student’s involvement
The student cannot touch/use animal materials without taking a training course at Kurume University School of Medicine.
Type of students accepted
This project accepts: - Medical students
- Chitinase 3-like-1 exacerbates intestinal inflammation by enhancing bacterial adhesion and invasion in colonic epithelial cells. Gastroenterology 2006;130:398.
- Chitinase 3-like 1 (CHI3L1/YKL-40) expression in colonic epithelial cells as a potentially novel marker for colitis-associated neoplasia. Am J Pathol 2011;179:1494.
- Chitin-binding domains of Escherichia colichiA mediate interactions with intestinal epithelial cells in mice with colitis. Gastroenterology 2013;145:602
- Novel methyxanthine derivative-mediated anti-inflammatory effects in inflammatory bowel disease. World Journal of Gastroenterology; 2014;20:1127.
- Chitinase 3-like 1 induces survival and proliferation of intestinal epithelial cells during chronic inflammation and colitis-associated cancer by regulating S100A9. Oncotarget; 2015;6:36535.
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