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

• 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
• 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

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世纪出现的重大健康挑战，影响 全世界有数百万的人，发病率上升。当前的疗法不足以诱导长期 许多患者的期限缓解，导致手术干预率很高，大肠风险增加 癌症（CRC）。因此，需要新的疗法来降低IBD的发病率和死亡率。许多患者 与IBD有关饮食纤维，短链脂肪酸（SCFA）的微生物代谢不足，因此 SCFA代表了治疗发展的潜在目标。 SCFA在肠内稳态中扮演复杂的角色 包括上皮周转，增殖，凋亡和免疫调节。 SCFA具有深远的影响 关于人类健康和疾病以及饮食纤维高的饮食与CRC的风险降低有关 IBD。迄今为止，使用SCFA作为IBD治疗的证据一直不一致。这些 不一致很可能是由于SCFA通过抑制组蛋白调节基因表达的能力而驱动的 脱乙酰基酶；但是，哪些基因受差异调节在很大程度上取决于SCFA的类型 相对于其他SCFA和细胞类型的浓度，所有这些都进一步使治疗复杂化。迄今为止， 由SCFA差异调节以诱导伤口愈合和调节的特定基因的身份 炎症是未知的。最近，我们确定了一种分泌的糖蛋白，牛奶脂肪球-EGF因子8 （MFGE8），它被SCFA显着上调。 MFGE8缺乏IBD患者，并且在 细胞迁移和T细胞分化。因此，该项目的长期目标是了解 SCFA在促进迁移和抑制炎症，伤口愈合中的关键过程和 保护结肠炎。我们的假设是SCFA差异调节肠上皮细胞 （IEC）MFGE8的表达以诱导伤口愈合并抑制IBD的炎症。使用MFGE8 缺乏细胞系和KO小鼠，我们将采用几种分子生物学技术，包括伤口愈合 测定，免疫组织化学，共聚焦显微镜，激酶测定，例如。体内肠培养基，流式细胞仪， 以及实验性结肠炎的急性和转移模型，通过两个特定目的解决我们的假设：1） 定义MFGE8在SCFA诱导的伤口愈合中的作用； 2）确定IEC的SCFA条件至 通过诱导MFGE8诱导Treg的发展。阐明MFGE8在SCFA诱导中的作用 IBD中伤口愈合和抑制炎症的抑制将有助于加深我们对 SCFA作为治疗性。这些信息将用于告知未来的治疗发展，涉及 SCFA，并最终消除了一个关键障碍，该障碍阻止了大型长期持续缓解 IBD患者的百分比。 呢