Microbiota metabolites SCFA promote intestinal epithelial repair and wound healing

微生物代谢物 SCFA 促进肠上皮修复和伤口愈合

基本信息

批准号：
9760793
负责人：
Anthony J Bilotta
金额：
$ 3.36万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-01 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9760793
关键词：
Acetates Acute Address Affect Anti-inflammatory Apoptosis Apoptotic Attenuated Biological Assay Butyrates CRISPR/Cas technology Cell Differentiation process Cell Line Cells Clinical Clinical Trials Colectomy Colitis Colorectal Cancer Complex Confocal Microscopy Data Dendritic Cells Development Diet Dietary Fiber Disease Disease remission EGF gene Epithelial Epithelial Cells Experimental Models FOXP3 gene Fermentation Flow Cytometry Future Gene Expression Genes Glycoproteins Goals Health Histone Deacetylase Inhibitor Homeostasis Human Immunohistochemistry In Vitro Incidence Inflammation Inflammatory Bowel Diseases Intestines Knockout Mice Knowledge Lead Link Modeling Molecular Biology Techniques Morbidity - disease rate Mucous Membrane Mus Natural regeneration Necrosis Operative Surgical Procedures Pathogenesis Pathway interactions Patients Phagocytosis Phenotype Phosphotransferases Play Process Production Propionates Proteins Proto-Oncogene Proteins c-akt Public Health Quantitative Reverse Transcriptase PCR Recombinants Regulation Regulatory T-Lymphocyte Research Project Grants Risk Role Stem cells T cell differentiation Therapeutic Up-Regulation Volatile Fatty Acids Western Blotting Wound Healing cell motility cell type colorectal cancer risk conditioning cytokine gut microbiota immunoregulation in vitro Assay in vivo intestinal homeostasis macrophage microbial microbiota metabolites migration milk expression milk fat globule mortality new therapeutic target novel novel therapeutics pre-clinical research prevent receptor repaired targeted treatment therapeutic development therapeutic target

项目摘要

ABSTRACT Inflammatory bowel disease (IBD) is a sizable health challenge that has emerged in the 21st century, affecting millions of people worldwide with incidence rates on the rise. Current therapies are insufficient to induce long- term remission in many patients, leading to high rates of surgical intervention and increased risk of colorectal cancer (CRC). Thus, new therapies are needed to decrease morbidity and mortality from IBD. Many patients with IBD are deficient in the microbial metabolites of dietary fibers, short chain fatty acids (SCFAs), and thus SCFAs represent a potential target for therapeutic development. SCFAs play complex roles in gut homeostasis including epithelial turnover, proliferation, apoptosis, and immune regulation. SCFAs have a profound impact on human health and disease, and a diet high in dietary fibers has been linked to decreased risk of CRC and IBD. To date, the evidence for use of SCFAs as a treatment for IBD has been inconsistent. These inconsistencies are most likely driven by SCFAs ability to regulate gene expression by inhibition of histone deacetylases; however, which genes are differentially regulated is largely dependent on the type of SCFA, its concentration relative to other SCFAs, and the cell type, all of which further complicate treatment. To date, the identity of the specific genes that are differentially regulated by SCFAs to induce wound healing and modulate inflammation are unknown. Recently, we identified a secreted glycoprotein, milk fat globule-EGF factor 8 (MFGE8), which is significantly upregulated by SCFAs. MFGE8 is deficient in IBD patients, and plays roles in cellular migration and T cell differentiation. Thus, the long-term goal of this project is to understand the role of SCFAs in the promotion of migration and suppression of inflammation, key processes in wound healing and protection against colitis. Our hypothesis is that SCFAs differentially regulate intestinal epithelial cell (IEC) expression of MFGE8 to induce wound healing and suppress inflammation in IBD. Using MFGE8 deficient cell lines and KO mice, we will employ several molecular biology techniques including wound healing assays, immunohistochemistry, confocal microscopy, kinase assays, ex. vivo enteroid cultures, flow cytometry, and acute and transfer models of experimental colitis to address our hypothesis through two specific aims: 1) Define the role of MFGE8 in SCFA-induced wound healing; 2) Determine SCFA conditioning of IEC to induce Treg development through induction of MFGE8. Elucidating the roles of MFGE8 in SCFA induction of wound healing and suppression of inflammation in IBD will help deepen our understanding of the potential of SCFAs as a therapeutic. This information will be utilized to inform future therapeutic development involving SCFAs, and ultimately remove a critical barrier that is preventing long-term sustained remission in a large percentage of IBD patients. !

抽象的炎症性肠病 (IBD) 是 21 世纪出现的一项巨大的健康挑战，影响着人们全球数百万人的发病率呈上升趋势。目前的疗法不足以诱导长期许多患者出现足月缓解，导致手术干预率较高，结直肠癌风险增加癌症（结直肠癌）。因此，需要新的疗法来降低 IBD 的发病率和死亡率。很多病人患有 IBD 的人缺乏膳食纤维、短链脂肪酸 (SCFA) 的微生物代谢产物，因此 SCFA 代表了治疗开发的潜在目标。 SCFA 在肠道稳态中发挥着复杂的作用包括上皮细胞更新、增殖、凋亡和免疫调节。 SCFA 具有深远的影响膳食纤维含量高的饮食与降低结直肠癌和癌症的风险有关炎症性肠病。迄今为止，使用 SCFA 治疗 IBD 的证据并不一致。这些不一致很可能是由 SCFA 通过抑制组蛋白调节基因表达的能力引起的脱乙酰酶；然而，哪些基因受到差异性调控很大程度上取决于 SCFA 的类型、其相对于其他 SCFA 的浓度以及细胞类型，所有这些都使治疗变得更加复杂。迄今为止，识别受 SCFA 差异调节以诱导伤口愈合和调节的特定基因炎症情况不明。最近，我们鉴定出一种分泌性糖蛋白——乳脂肪球——EGF因子8 (MFGE8)，它被短链脂肪酸 SCFA 显着上调。 MFGE8 在 IBD 患者中存在缺陷，并在以下方面发挥作用：细胞迁移和 T 细胞分化。因此，该项目的长期目标是了解 SCFAs 促进迁移和抑制炎症、伤口愈合的关键过程以及预防结肠炎。我们的假设是 SCFA 差异性调节肠上皮细胞 (IEC) MFGE8 表达可诱导 IBD 伤口愈合并抑制炎症。使用 MFGE8 缺陷细胞系和 KO 小鼠，我们将采用多种分子生物学技术，包括伤口愈合测定、免疫组织化学、共聚焦显微镜、激酶测定等。体内肠样培养、流式细胞术、以及实验性结肠炎的急性和转移模型，通过两个具体目标来解决我们的假设：1）定义 MFGE8 在 SCFA 诱导的伤口愈合中的作用； 2) 确定 IEC 的 SCFA 条件通过诱导 MFGE8 诱导 Treg 发育。阐明 MFGE8 在 SCFA 诱导中的作用 IBD 中伤口愈合和炎症抑制的研究将有助于加深我们对 IBD 潜力的了解 SCFA 作为治疗剂。该信息将用于指导未来的治疗开发，包括 SCFA，最终消除阻碍大规模长期持续缓解的关键障碍 IBD 患者的百分比。！