权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Novel cytoskeletal mechanisms of pathogenic bacteria interactions with intestinal epithelium

病原菌与肠上皮相互作用的新细胞骨架机制

基本信息

批准号：
10663379
负责人：
Andrei Ivanovich Ivanov
金额：
$ 50.68万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10663379
关键词：
Acceleration Actins Adherent Invasive Escherichia coli Antibiotic Therapy Attenuated Automobile Driving Bacteria Bacterial Adhesion Bacterial Antibiotic Resistance Bacterial Attachment Site Bacterial Infections CRISPR/Cas technology Cell Communication Cell Line Cell surface Cells Chemicals Chronic Citrobacter rodentium Cytoskeleton Data Disease Economic Burden Elderly Enteral Epithelial Cells Epithelium Escherichia coli Escherichia coli EHEC Event Filament Gastroenteritis Gastrointestinal Diseases Genes Genetic Health Hemolytic-Uremic Syndrome Human Immunocompromised Host In Vitro Infant Infection Inflammatory Inflammatory Bowel Diseases Intestinal Diseases Intestinal Mucosa Invaded Knock-out Knockout Mice Mediating Microfilaments Modeling Molecular Molecular Chaperones Morbidity - disease rate Motor Motor Activity Mus Myosin ATPase Myosin Type II Organoids Outcome Pathogenesis Pathogenicity Pathway interactions Patients Persons Play Polymers Prevention Process Protein Isoforms Proteins Public Health RNA Interference Regulation Role Route Salmonella Salmonella enterica Severities Shigella Symptoms Systemic disease Testing Therapeutic Intervention Transgenic Mice blebbistatin cohort crosslink enteric infection enteric pathogen enteropathogenic Escherichia coli gastrointestinal genetic approach genetic manipulation gut colonization improved in vivo inhibitor innovation insight intestinal epithelium monolayer mortality mutant new therapeutic target non-muscle myosin novel novel therapeutic intervention novel therapeutics pathogen pathogenic bacteria pharmacologic polymerization receptor

项目摘要

ABSTRACT Intestinal Epithelial Cells (IEC) provide the first line of defense against enteric pathogenic bacteria. However bacterial pathogens developed different strategies to colonize intestinal epithelium causing severe gastrointestinal disorder. One strategy, used by ‘attaching and effacing’ bacteria (AEB), such as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC), involves bacterial adhesion to IEC without entering the host cells. The other strategy used by adherent Invasive E. coli (AIEC) or Salmonella results in bacterial internalization into IEC. These different colonization routes bear a key mechanistic similarity: they require rearrangements of the epithelial actin cytoskeleton to enable bacterial attachment and invasion. One important but understudied mechanism of pathogen-induced cytoskeletal remodeling involves a major actin cytoskeleton motor, non-muscle myosin II (NM-II). Our preliminary data demonstrate a striking dualism of NM-II-dependent regulation of bacterial-IEC interactions. Pharmacologic and genetic inhibition of NM-II increases AEB attachment to IEC, but inhibits AIEC invasion in vitro and in vivo. These contrasting roles of NM-II in IEC-bacterial interactions likely reflect exploiting two different NM II-dependent functions: its actin filament contractility and actin filament cross-linking activity, by different pathogens. This exciting data provides a scientific premise for the following innovative hypothesis: NM-II plays a dual role in regulating enteric pathogen interactions with intestinal epithelium by inhibiting AEB attachment to epithelial cells, but promoting epithelial entry of invading pathogenic bacteria. This hypothesis will be tested in the following Aims: (1) to determine the roles of NM-II in regulating intestinal epithelial cell interactions with attaching and effacing bacteria; (2) to dissect the mechanisms of NM-II-driven internalization of invading bacteria into intestinal epithelial cells. We will study EPEC, EHEC, AIEC and Salmonella colonization of model IEC cell lines, organoid-derived IEC monolayers in vitro and intestinal colonization by Citrobacter rodentium, AIEC and Salmonella in vivo. NM-II activity will be modulated by pharmacologic and genetic approaches. The genetic approach will target two major epithelial NM-II isoforms, NM-IIA and NM-IIC, by selectively downregulating their expression in human IEC using CRISPR/Cas9 gene editing and RNA interference. In vivo NM-II functions will be examined by using mice with either intestinal epithelial specific knockout of NM-IIA, or total knockout of NM-IIC. To determine which NM-II activity is essential for bacterial-IEC interactions, we will utilize IEC cells and transgenic mice expressing NM-IIA mutants deficient in either actin filament cross-linking, or filament contraction activities. Significance: the proposal will reveal novel insights into understanding how the intestinal epithelium controls pathogenic bacterial infections and will identify new targets for therapeutic interventions to treat diseases caused by enteric pathogens.

摘要