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Distal gut microbiome targets of host anti-proteolytic proteins during colitis

结肠炎期间宿主抗蛋白水解蛋白的远端肠道微生物组目标

基本信息

批准号：
10320030
负责人：
Howard C Hang
金额：
$ 22.19万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-12-18 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10320030
关键词：
Abate Address Affinity Algorithms Animal Model Anti-Bacterial Agents Anti-Inflammatory Agents Antiinflammatory Effect Binding Binding Proteins Bioinformatics Biological Biology C57BL/6 Mouse Cells Chemicals Chronic Colitis Collaborations Collection Colorectal Cancer Communities Control Groups Cysteine Data Development Disease Distal Drug Targeting Elastases Enzymes Experimental Designs Family Female Future Genetic Genomics Goals Hand Harvest Homeostasis Human Ileitis Immune Immunocompromised Host In Vitro Infiltration Inflammation Inflammatory Injections Intestinal Content Intestinal Mucosa Intestines Knowledge Laboratories Letters Libraries Liquid Chromatography Maps Mass Spectrum Analysis Measures Metabolism Methodology Methods Modeling Mus Obesity PI3 gene Parents Patients Peptide Hydrolases Property Proteins Proteome Proteomics Publishing Rag1 Mouse Research Role Sampling Serine Serine Protease T cell therapy T-Lymphocyte Technology Time Translations Trypsin antimicrobial base biochemical model chemically induced colitis commensal bacteria drug discovery gut inflammation gut microbiome in vivo interest metaproteomics microbial microbial host microbiome mouse model murine colitis pathogenic bacteria response small molecule stem tandem mass spectrometry therapeutic target

项目摘要

Title: Distal gut microbiome targets of host anti-proteolytic proteins during colitis ABSTRACT Our long-term goal is to advance our understanding of the distal gut microbiome from its basis in genomics with chemical biology and quantitative metaproteomics approaches. Over the last several years, we have developed a combination of chemical probe-based technologies, liquid chromatography tandem mass spectrometry (LC-MS/MS) proteomics, and bioinformatic analyses as approaches to interrogate and quantitate specific enzyme families in microbiome proteomes. We recently showed that colitic mice have a dramatic elevation in microbiome-secreted proteases within the distal gut relative to healthy mice using these combined methods. Correspondingly, we observe a significant increase in host anti-proteolytic proteins (APPs). Gut inflammation can be abated in colitis mouse models upon introduction of human APPs a-1-antitrypsin or elafin. While we posit that a-1-antitrypsin and elafin (and other APPs) reduce inflammation via inhibition of aberrant proteolytic activities, both APPs also have antimicrobial activity. Thus, the anti-inflammatory mechanism by which antitrypsin and elafin act could be primarily, or in part, attributable to altering bacterial composition. Our primary question is if APPs target microbial proteases. Here, we will elucidate the microbiome (and host) protein targets of six APPs overproduced in response to distal gut inflammation. These six APPs collectively target proteases from the serine, cysteine, and metallo subfamilies. We have purified all APPs of interest and aim to employ the proteins as “bait” to determine if these APPs primarily target the inhibition of bacterial proteases. We will subsequently identify and quantitate the APP-captured proteins with our LC-MS/MS metaproteomics methods. We propose to focus our APP-enrichment strategy and metaproteomics on a well- established adoptive T cell transfer murine model of chronic colitis and directly compare to control mice raised under identical conditions. Our preliminary data confirms that a-1-antitrypsin irreversibly binds a unique collection of secreted proteins within colitic microbiome samples compared to controls. This is significant, as expression levels of host elastase and trypsin (targets of a-1-antitrypsin) are similar between the colitis and control groups. Over the next two years, we anticipate that we will identify several families of host APP-targeted bacterial proteases upregulated in colitic mice, as classified by sequence and/or structural homology. This list of proteases will help guide and focus future biochemical, cell-based, and animal model research on specific protease families as potential inflammatory agents (and therapeutic targets), as well as provide an optimized APP-enrichment methodology for translation into human IBD patients and other microbiome-related diseases.

题目：结肠炎期间宿主抗蛋白水解蛋白的远端肠道微生物靶点