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

TRPM2 mediates neutrophil transendothelial migration and inflammation

TRPM2介导中性粒细胞跨内皮迁移和炎症

基本信息

批准号：
9260918
负责人：
CHINNASWAMY TIRUPPATHI
金额：
$ 39.98万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-15 至 2019-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9260918
关键词：
Address Adherens Junction Adhesions Bacteria Bacterial Infections Blood Vessels CRISPR/Cas technology Calcium Cell Communication Cell surface Cell-Cell Adhesion Cells Complex Data Dissociation Endothelial Cells Endothelium Event Extravasation Generations Guide RNA Host Defense Immunity Infection Infiltration Inflammation Inflammatory Inflammatory Response Innate Immune Response Invaded Knock-out Lead Leukocyte Rolling Leukocytes Lipopolysaccharides Lung Lung Inflammation Mediating Modeling Molecular Genetics Mus NADP Nuclear Oxidants Oxidases P-Selectin PRKCA gene Phase Phosphorylation Physiological Play Polymorph Production Pseudomonas aeruginosa Reactive Oxygen Species Recruitment Activity Role Route SRC gene Secondary to Seminal Signal Transduction Site System Testing Therapeutic Time Tissues Toxin Transgenic Mice cadherin 5 defense response improved killings leucyl-phenylalanine leukocyte activation microorganism migration mouse model neutrophil novel pathogen public health relevance receptor response sulfated glycoprotein 2 trafficking

项目摘要

DESCRIPTION (provided by applicant): Reactive oxygen species (ROS) produced by polymorph nuclear leukocytes (PMNs) and endothelial cells (ECs) during the innate immune response against invading pathogens are of major importance in governing PMN capture and extravasation at a site of infection. Although we have demonstrated the involvement of the transient receptor potential melastatin (TRPM) channel 2 (TRPM2) in mediating oxidant-induced calcium entry and loss of the endothelial barrier integrity in endothelial cells (ECs), the functio of the TRPM2 channel in ROS-induced trans endothelial migration of PMNs have not been assessed. Our studies suggest that TRPM2 functions in response to ROS to activate vascular PMN extravasation in infected lungs. We will thus test the hypothesis that the increase in lung micro vessel PMN transmigration and clearance of the bacterial pathogens from the infected lungs depend on the activation of endothelial TRPM2 by oxidants. To test this hypothesis we will follow the three specific aims: Aim #1 will address the requirement of the endothelial cell-TRPM2 channel in mediating transendothelial migration of PMNs at sites of bacterial infection, thereby promoting clearance of the invading pathogen from the lung. The proposed studies will use a transgenic mouse model expressing endothelial cell- restricted deletion of TRPM2 and mouse lung endothelia transduced with TRPM2 crispr/Cas9-gRNA to address the role of TRPM2 activation in the mechanism of increased lung PMN transmigration. We will establish the functional significance of TRPM2 in signaling PMN activation-induced lung PMN infiltration and bacterial clearance in a murine model of Pseudomonas aeruginosa pulmonary infection. Aim #2 will address whether PMN-activated TRPM2 plays a critical role during bacterial infection by distributing P-selecting on the EC surface, and recruiting PMNs to the site of infection. Specifically, we will determine whether TRPM2-regulated Ca2+ entry induces P-selectin mobilization to EC surface through activation of PKCa signaling. Aim #3 will address the signaling mechanisms by which PMN-induced activation of TRPM2 in ECs mediates formation of vascular interendothelial gaps and PMN transendothelial migration. The studies will establish the role of TRPM2-regulated Ca2+ signaling in ECs in promoting disassembly of Adherens Junctions complexes through phosphorylation of the vascular endothelial (VE)-cadherin, resulting in loss of endothelial cell-cell adhesion and thereby mediating PMN transmigration. Overall, these studies will establish the functional significance of TRPM2 in the induction of PMN transendothelial migration, which is necessary for host defense against bacterial infections; therefore, manipulating TRPM2 function in the endothelium represents a novel target to mount effective host's immunity against pulmonary gram-negative infection.