To Explore the Mechanism of Probiotics Bacteria and Extracellular Vesicles of L. Casei Rhamnosus to Modulate Host Regulatory T Cells and Microbiota against Colitis and Colon Cancer

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Project title

Code/計畫編號

MOST109-2320-B031-001

Translated Name/計畫中文名

探討益生菌及胞外泌體藉由宿主調節性Ｔ細胞及腸道菌相以改善腸炎及大腸癌之相關機制

Funding Organization/主管機關

National Science and Technology Center for Disaster Reduction

Co-Investigator(s)/共同執行人

邱亦涵(計畫主持人)

Department/Unit

St. Mary's Junior College of Medicine, Nursing and Management College

Website

https://www.grb.gov.tw/search/planDetail?id=13550689

Year

2020

Start date/計畫起

01-01-2020

Expected Completion/計畫迄

31-07-2021

Co-Investigator(s)

Chang-Jer Wu

Bugetid/研究經費

1000千元

ResearchField/研究領域

食品科技(工)

根據衛生署的調查，大腸癌發生率連續十年盤踞台灣癌症的第一位。而發炎性腸道疾病為一種慢性反覆發炎的免疫疾病，是演進成大腸癌的重要原因之一。發炎性腸道疾病發生時，腸道免疫失調，CD4+T細胞極化轉變成Th1和T17細胞比例增加，而調節型T細胞 (Tregs) 減少，身體的免疫系統對腸道發生攻擊，同時增加發炎細胞激素的生成而造成發炎，致使大腸癌發生的危險性增加。我們過去的研究發現，L. casei rhamnosus所分泌的胞外物質可促使活化免疫細胞的凋亡並抑制發炎細胞激素的生成; 而本計畫書內容所進行的先驅實驗結果亦顯示，L. casei rhamnosus所分泌的胞外物質可於體外誘導調節型Ｔ細胞的增生，並顯著降低DSS所誘導潰瘍性結腸炎的腸道損傷。因此，本研究將以三種大腸炎到大腸癌轉型模式，分成三年期計畫探討L. casei rhamnosus菌體及胞外泌體對腸道菌相、腸道損傷和免疫細胞極化之調控機制。各年度之研究目標分述於下: 第一年: 分為兩個主題探討：(一) 以細胞模式探討L. casei rhamnosus菌體及胞外泌體抗發炎及誘導調節性Ｔ細胞增生的可能機轉；(二) 進行L. casei rhamnosus胞外泌體蛋白質譜儀定量及系統生物分析。第二年: 分為兩個主題探討：(一) 以DSS引致小鼠急性腸炎，探討L. casei rhamnosus菌體及胞外泌體對於調節性T細胞活化、腸道發炎反應、組織損傷及腸道菌相之影響，了解L. casei rhamnosus菌體及胞外泌體緩解腸炎之保護效應；(二) 以AOM/DSS引致小鼠發炎性大腸癌早期模式，探討L. casei rhamnosus菌體及胞外泌體對於發炎性大腸癌模式小鼠腸道菌相、腫瘤形成、腫瘤嚴重性、細胞凋亡及增生之影響。第三年:探討L. casei rhamnosus菌體及胞外泌體對AOM/DSS引致小鼠發炎性大腸癌晚期模式的腸道發炎反應、組織損傷及腸道菌相及抗腫瘤免疫之影響，並運用基因體學、蛋白質體學、組織染色技術、免疫技術探討相關基因表現及細胞訊息傳遞路徑。 According to the latest cancer incidence report by Department of Health, colorectal cancer continued to be the cancer affecting the highest number of Taiwanese people for recent 10 consecutive years. Colitis associated cancer (CAC) is the type of colon cancer which is preceded by clinically detectable inflammatory bowel disease (IBD). IBD is a relapsing and remitting disorder characterized by chronic inflammation of the gastrointestinal tract. Deregulation of immune responses in the gut plays a critical role in IBD. During IBD, naïve CD4+ T cells can be activated and polarized subsequently into Th1/Th17 immune response, while that is related with a decrease in the number of regulatory T cells (Tregs) and an increase in the secretion of pro-inflammatory cytokines, leads to malignant transformation of colon. Our previous study demonstrated probiotic L. casei rhamnosus (Lcr) produces heat-stable soluble molecules to promote active immune cell apoptosis without affecting intestinal epithelial cells. And the soluble molecules of Lcr also inhibit LPS-induced inflammatory cytokines by immune cells. Meanwhile, our preliminary study demonstrated that Lcr supernatant significant induce Tregs differentiation. And, oral administration of Lcr culture supernatant significantly attenuates intestinal epithelial barrier injury and inflammatory responses in dextran sodium sulfate (DSS)-induced acute colitis of mice. Therefore, we propose to carry out this study in three consecutive years to clarify the effects of Lcr probiotics bacteria (PB) and Lcr derived extracellular vesicle (LDEVs) on intestinal injury, CD4+ T-cell polarization and gut microbiota on the acute colitis murine model, and colitis-associated cancer murine model in the early and late phases. The aim of each year is listed below: The first year: Experiment 1: Using in vitro cell model to explore the mechanism of PB and LDEVs in anti-inflammation and Tregs differentiation. Experiment 2: Gene ontology analysis of Lcr EV proteome. The second year: Experiment 1: Using in vivo DSS-induced acute colitis murine model to investigate the effects of PB and LDEVs on gut microbiota, intestinal damage and inflammation, and to explore whether PB and LDEVs modulate CD4+ T-cell polarization to Tregs differentiation. Experiment 2: An azoxymethane (AOM)/DSS-induced colitis-associated cancer murine model in the early phase will be used to investigate the effects of PB and LDEVs on gut microbiota, intestinal damage and inflammation, and to explore the role of PB and LDEVs during the course of IBD predisposition to colon cancer. The third year: An AOM/DSS-induced colitis-associated cancer murine model in the late phase will be used to investigate the effects of PB and LDEVs on gut microbiota, intestinal damage and inflammation, and to examine the mechanism involved in the PB and LDEVs associated attenuation against colitis predisposed colorectal cancer. The related gene expression and cellular pathway will be determined using the genomic, proteomic, immuno-histological and immunological techniques. Overall, the outcome of this project would be valuable in decreasing the IBD predisposed colon cancer.

Keyword(s)

益生菌
胞外泌體
調節型T細胞
腸道菌相
潰瘍性腸炎
大腸癌
probiotics
extracellular vesicles
regulatory T cell
gut microbiota
colitis
colon cancer

DSpace CRIS

To Explore the Mechanism of Probiotics Bacteria and Extracellular Vesicles of L. Casei Rhamnosus to Modulate Host Regulatory T Cells and Microbiota against Colitis and Colon Cancer

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