Regulation of protein secretion in human gut commensals

人类肠道共生体蛋白质分泌的调节

• 批准号: 10550129
• 负责人: Brent W Anderson
• 金额: $ 6.95万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10550129
• 关键词: Address; Adult; Amines; Bacteria; Bacterial Proteins; Bacteroides; Bacteroides fragilis; Bacteroides thetaiotaomicron; Bacteroidetes; Biological Assay; Biomedical Engineering; Career Mobility; Caspase; Cells; Colon Carcinoma; Communication; Complex; Cues; Digestion; Educational process of instructing; Environment; Enzymes; Faculty; Food; Fostering; Genetic; Genetic Determinism; Gnotobiotic; Goals; Growth; Harvest; Health; Homologous Gene; Human; Intestines; Invaded; Isotope Labeling; Learning; Link; Measures; Mediating; Mentors; Mentorship; Microbe; Microbiology; Mus; Nutrient; Nutrient availability; Organism; Outcome; Pathway interactions; Peptide Hydrolases; Phase; Postdoctoral Fellow; Process; Proliferating; Protein Secretion; Proteins; Proteome; Proteomics; Regulation; Research; Research Personnel; Resistance; Role; Science; Signal Transduction; Signal Transduction Pathway; Surface; Techniques; Testing; Time; Toxin; Training; Universities; Western Blotting; Work; bacterial community; bacteriocin; career development; commensal bacteria; differential expression; gut bacteria; gut microbes; gut microbiome; gut microbiota; host microbiome; human disease; in vivo; innovation; microbial; microbial community; microbiome; mouse model; multidisciplinary; novel; pathogen; peer; response; secretion process; skills; student mentoring; symposium; therapeutic enzyme

Project Summary Commensal bacteria in the human intestinal tract provide numerous benefits to their hosts, from digestion of foods inaccessible to the host to protection against pathogen invasion. Many host-microbiome interactions are mediated by bacterial proteins secreted into the gut (the secretome), but regulation and coordination of this process has not been studied. This proposal focuses on the dominant gut genus Bacteroides to test the hypothesis that these organisms use central mechanisms to control deployment of secreted proteins. One clue comes from studies of commensal-encoded toxins: two secreted toxins with no similarities in sequence or targets both use the same surface-associated cysteine protease to orchestrate their secretion. Homologs of this cysteine protease (but not its targets) are conserved across Bacteroides, suggesting a broader role for these enzymes in regulating protein secretion. This proposal will use an innovative N-terminome proteomic approach to determine the repertoire of protease substrates in Bacteroides and establish whether protease homologs share targets within and between species. Complementary studies in gnotobiotic mice will determine the contribution of protease processing to the in vivo secretome. This proposal will also use genetics, anaerobic microbiology, and gnotobiotics to elucidate environmental cues and signal transduction pathways that regulate protease expression. This will determine how these proteases might coordinate release of waves of secreted proteins in response to environmental signals in the gut. If successful, these studies will uncover fundamental mechanisms for secretome regulation in the microbiome, with implications for host-microbiome interactions and bioengineering therapeutic enzyme delivery in the gut. This proposal also presents a detailed training plan that will allow the candidate to develop new skills required for career advancement. This research will be conducted at the highly collaborative and multidisciplinary Microbial Sciences Institute (MSI) at Yale University. The sponsor has an established track record of mentorship, and the lab and the MSI provide state-of-the-art facilities. The diverse expertise of peers and faculty at MSI, ranging from chemists, biochemists, and geneticists, will support the candidate's research goals. A mentoring committee will provide technical guidance for the project and will support the candidate's career development. The candidate will also complete training in mentoring and teaching from the Poorvu Center for Teaching and Learning and the Yale Postdoctoral Association, and he will put this training into practice by mentoring students in their local science fair projects. By presenting his work to peers at Yale University and in conferences, the candidate will foster his communication skills and build a network of colleagues. Together, this training plan will build the skillset and connections required to succeed as an independent investigator.

项目摘要 人类肠道中的共生细菌为其宿主提供了许多好处，从消化中 宿主无法访问的食物可以防止病原体入侵。许多宿主 - 微生物组相互作用是 由分泌到肠道的细菌蛋白介导（分泌组），但调节和协调 过程尚未研究。该提案重点介绍了主要的肠道菌属，以测试 假设这些生物使用中心机制来控制分泌蛋白的部署。一个线索 来自对共生编码的毒素的研究：两个分泌的毒素，没有序列或没有相似性或 靶标都使用相同的表面相关的半胱氨酸蛋白酶来编排它们的分泌。同源物 这种半胱氨酸蛋白酶（但不是其靶标）在整个细菌中保存，这表明对 这些酶在调节蛋白质分泌方面。该建议将使用创新的N末端蛋白质组学 确定细菌蛋白酶底物的曲目的方法，并确定蛋白酶是否蛋白酶 同源物在物种之间和物种之间共享目标。 gnotobiotic小鼠的互补研究将确定 蛋白酶加工对体内分泌组的贡献。该建议还将使用遗传学， 厌氧微生物学和gnotobiotics阐明环境线索和信号转导途径 调节蛋白酶表达。这将确定这些蛋白酶如何协调波的释放 分泌的蛋白质响应于肠道中的环境信号。如果成功，这些研究将发现 微生物组中分粒子调节的基本机制，对宿主微生物组的影响 肠道中的相互作用和生物工程治疗酶递送。 该建议还提出了详细的培训计划，该计划将使候选人能够发展新技能 职业发展所必需的。这项研究将在高度协作和 耶鲁大学的多学科微生物科学研究所（MSI）。赞助商有一个既定的曲目 指导记录，实验室和MSI提供了最先进的设施。同行的多样化专业知识 MSI的教师（从化学家，生物化学家和遗传学家）都将支持候选人的研究 目标。指导委员会将为该项目提供技术指导，并将支持候选人的 职业发展。候选人还将完成Podvu的指导和教学培训 教学中心和耶鲁大学博士后协会，他将进行此培训 通过指导学生在当地科学博览会项目中进行练习。通过向耶鲁大学的同行介绍他的作品 大学和会议上，候选人将促进他的沟通技巧，并建立一个网络 同事。该培训计划将共同建立成功的技能和联系 独立研究者。