Metaproteomics to investigate intestinal microbiota-host and -diet interactions

宏蛋白质组学研究肠道微生物群-宿主和-饮食相互作用

• 批准号: 10406980
• 负责人: Manuel Kleiner
• 金额: $ 37.02万
• 依托单位: NORTH CAROLINA STATE UNIVERSITY RALEIGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10406980
• 关键词: Biochemical; Biological Assay; Communities; Complement; Consumption; Custom; Development; Diet; Dietary Proteins; Disease; Environment; Foundations; Gene Expression; Goals; Health; Human; Human Microbiome; In Vitro; Intestines; Knowledge; Laboratory Study; Mass Spectrum Analysis; Measurement; Metagenomics; Methods; Mission; National Institute of General Medical Sciences; Physiology; Prevention; Proteins; Public Health; Reproducibility; Research; Resolution; Site; Source; Therapeutic Intervention; United States National Institutes of Health; base; cellular imaging; disease diagnosis; gut microbiota; host microbiota; host-associated microbial communities; human microbiota; insight; member; metabolic abnormality assessment; metabolomics; metaproteomics; microbial; microbial community; microbiota; novel strategies; rational design; response; therapeutic development; tool

Project Summary/Abstract In the Kleiner laboratory we study metabolism, physiology and interactions in microbial symbioses and host- associated microbiota. For this we combine a diversity of cultivation independent approaches – such as metagenomics, metabolomics, metaproteomics and single cell imaging – with cultivation-based approaches including heterologous gene expression, biochemical assays and other in vitro measurements. Our research has a strong focus on the development of high-resolution mass spectrometry driven metaproteomics for the large-scale identification and quantification of proteins in host-associated microbiota. During the next five years, I plan to continue using these approaches to study intestinal microbiota responses to, and mechanisms of interaction with, external substrates (diet) and host-derived substrates (host compound foraging). I hypothesize that different dietary protein sources and host-derived compounds will have vastly different impacts on the microbiota and thus need to be considered when studying the interconnection of diet, the microbiota, and host health. I propose to use metaproteomics, complemented with metagenomics and metabolomics, to (1) identify and quantify the substrates that are used and converted by microbiota members and (2) determine how these substrates impact community composition and functional interactions with other microbiota members and the host. This research will provide urgently needed insight into the functional impacts of substrates consumed by gut microbiota by optimizing and deploying novel approaches for the reproducible, large-scale characterization of host, diet and microbial proteins in the intestinal tract. My long-term goals are to develop metaproteomic approaches that allow us to quantitatively and reproducibly determine functional interactions in microbial communities, and to define critical interactions between the microbiota and dietary proteins that will inform the development of therapeutic interventions. These approaches will also be powerful tools for studying any disease associated with the human microbiome in and on different body sites and microbial communities that humans interact with in their environment and that potentially impact health.

项目概要/摘要 在凯鹏华盈实验室，我们研究新陈代谢、生理学以及微生物共生和宿主的相互作用。 为此，我们结合了多种独立于培养的方法，例如 宏基因组学、代谢组学、宏蛋白质组学和单细胞成像——采用基于培养的方法 包括异源基因表达、生化测定和其他体外测量。 重点关注高分辨率质谱驱动的宏蛋白质组学的开发 宿主相关微生物群中蛋白质的大规模鉴定和定量。 在接下来的五年中，我计划继续使用这些方法来研究肠道微生物群的反应 外部底物（饮食）和宿主衍生底物（主体化合物 我认为不同的膳食蛋白质来源和宿主衍生的化合物会有很大的差异。 对微生物群的不同影响，因此在研究饮食之间的相互关系时需要考虑， 我建议使用宏蛋白质组学，并辅以宏基因组学和 代谢组学，(1) 识别和量化微生物群成员使用和转化的底物 (2) 确定这些底物如何影响群落组成以及与其他底物的功能相互作用 这项研究将为了解微生物群成员和宿主的功能提供迫切需要的见解。 通过优化和部署新方法来研究肠道微生物群消耗的底物的影响 对肠道中宿主、饮食和微生物蛋白质进行可重复的大规模表征。 我的长期目标是开发宏蛋白质组学方法，使我们能够定量且可重复地 确定微生物群落中的功能相互作用，并定义微生物群落之间的关键相互作用 微生物群和膳食蛋白质将为治疗干预措施的发展提供信息。 方法也将成为研究与人类微生物组相关的任何疾病的有力工具。 人类在其环境中与之相互作用的不同身体部位和微生物群落 可能影响健康。