Determining the role of autophagy in regulating the Staphylococcus aureus a-toxin receptor ADAM10

确定自噬在调节金黄色葡萄球菌 a-毒素受体 ADAM10 中的作用

基本信息

批准号：
9762176
负责人：
Matthew David Keller
金额：
$ 3.58万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9762176
关键词：
Affect Amino Acid Motifs Antibiotic Resistance Attention Autophagocytosis Autophagosome Bacteria Bacterial Infections Bacterial Toxins Binding Biology Cell Adhesion Cell Death Cell Surface Receptors Cell membrane Cell physiology Cell surface Cells Clinical Collaborations Community Hospitals Community-Acquired Infections Cytolysis Data Degradation Pathway Dependence Development Disease Disintegrins Docking Elements Endothelial Cells Endothelium Event Family Future Generations Goals Homeostasis Hospitals Immune System Diseases Immune response In Vitro Incidence Infection Inflammatory Integral Membrane Protein Integration Host Factors Intervention Intravenous Life Lung Lysosomes Maintenance Mediating Membrane Membrane Potentials Membrane Proteins Metalloproteases Methicillin Methods Mus Organelles Pathogenesis Pathology Pathway interactions Play Predisposition Process Production Proteins Public Health Regulation Research Role Site Specificity Staphylococcus aureus Staphylococcus aureus infection Structure System Testing Tissues Toxin Vancomycin Vancomycin-resistant S. aureus Vesicle Virulence Virulence Factors alpha Toxin antimicrobial community setting cytotoxicity experimental study extracellular extracellular vesicles health care settings in vivo Model insight macromolecule member methicillin resistant Staphylococcus aureus multicatalytic endopeptidase complex novel novel therapeutics pathogen prevent receptor resistant strain respiratory therapeutic target trafficking

Affect Amino Acid Motifs Antibiotic Resistance Attention Autophagocytosis Autophagosome Bacteria Bacterial Infections Bacterial Toxins Binding Biology Cell Adhesion Cell Death Cell Surface Receptors Cell membrane Cell physiology Cell surface Cells Clinical Collaborations Community Hospitals Community-Acquired Infections Cytolysis Data Degradation Pathway Dependence Development Disease Disintegrins Docking Elements Endothelial Cells Endothelium Event Family Future Generations Goals Homeostasis Hospitals Immune System Diseases Immune response In Vitro Incidence Infection Inflammatory Integral Membrane Protein Integration Host Factors Intervention Intravenous Life Lung Lysosomes Maintenance Mediating Membrane Membrane Potentials Membrane Proteins Metalloproteases Methicillin Methods Mus Organelles Pathogenesis Pathology Pathway interactions Play Predisposition Process Production Proteins Public Health Regulation Research Role Site Specificity Staphylococcus aureus Staphylococcus aureus infection Structure System Testing Tissues Toxin Vancomycin Vancomycin-resistant S. aureus Vesicle Virulence Virulence Factors alpha Toxin antimicrobial community setting cytotoxicity experimental study extracellular extracellular vesicles health care settings in vivo Model insight macromolecule member methicillin resistant Staphylococcus aureus multicatalytic endopeptidase complex novel novel therapeutics pathogen prevent receptor resistant strain respiratory therapeutic target trafficking

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项目摘要

Project Summary S. aureus is responsible for a large number of infections in the community and healthcare setting. Especially as the number and incidence of antibiotic resistant strains continue to rise, the need for alternative intervention methods is becoming increasingly critical. A strategy to develop novel therapies is to identify and block host pathways exploited by pathogens to cause disease. Autophagy is one such pathway. Autophagy is a highly conserved, ubiquitous cellular process in which a double membrane autophagosome engulfs damaged cytosolic components and targets them for lysosomal degradation; however, recent research has demonstrated its critical role in pathogen tolerance and clearance. Experiments performed in collaboration between the Cadwell and Torres labs demonstrated a vital role for ATG16L1, a protein that mediates autophagosome formation, in S. aureus tolerance. Using an in vivo model of autophagy loss where ATG16L1 is almost completely abolished, the increased susceptibility to lethal challenge was found to be dependent on the production of the S. aureus virulence factor, -toxin. Upon further experimentation, it was shown that endothelial cells lacking ATG16L1 display higher levels of the plasma membrane -toxin receptor ADAM10. These data suggest autophagy plays a negative regulatory role on ADAM10. However, it is remains unclear how autophagy, a cytosolic degradation pathway, regulates the levels of membrane bound ADAM10. Our aim is to identify host factors, mechanisms, and/or pathways that are differentially regulated by autophagy that affect ADAM10 levels. Using an in vitro system that we developed for this purpose, our initial data suggests that ATG16L1 regulates ADAM10 independently of the lysosome or proteasome. Instead, cells lacking ATG16L1 show decreased production of extracellular vesicles containing ADAM10. We plan to continue to test precisely how ATG16L1 and autophagy influence native localization of ADAM10; particularly through packaging into vesicles meant for extracellular release, trafficking to the plasma membrane, and endocytic internalization. Additionally, our goal is to determine the amino acid motifs or structural elements of ADAM10 that confer its autophagy dependent regulation. Each one of these strategies is an attempt to better understand the biology of ADAM10 regulation by autophagy as this pathway and its substrates may serve as alternative targets for treatment of S. aureus infections and other conditions involving the endothelial barrier.

项目摘要金黄色葡萄球菌负责社区和医疗保健环境中的大量感染。特别是抗生素抗性菌株的数量和事件继续增加，需要替代干预方法变得越来越关键。开发新疗法的策略是识别和阻止宿主病原体探索的途径引起疾病。自噬就是这样的途径。自噬是一个高度保守的无处不在的细胞过程，其中双膜自噬体吞噬了胞质组件并将其靶向溶酶体降解；但是，最近的研究表明了它的关键在病原体耐受性和清除率中的作用。在Cadwell和Torres Labs之间合作进行的实验表明了至关重要的作用对于ATG16L1，在金黄色葡萄球菌耐受性中介导自噬体形成的蛋白质。使用体内模型 ATG16L1几乎完全废除的自噬损失，对致命的敏感性增加发现挑战取决于金黄色葡萄球菌病毒因子茶素的产生。进一步实验表明，缺乏ATG16L1的内皮细胞显示出更高水平的等离子体膜毒素受体ADAM10。这些数据表明自噬对 ADAM10。但是，尚不清楚自噬如何调节胞质降解途径膜绑定的ADAM10。我们的目的是确定宿主因素，机制和/或途径不同的途径。影响ADAM10水平的自噬。使用我们为此目的开发的体外系统，我们的初始系统数据表明ATG16L1独立于溶酶体或蛋白酶体来调节ADAM10。相反，细胞缺乏ATG16L1显示含有ADAM10的蔬菜的产生减少。我们计划继续精确测试ATG16L1和自噬如何影响ADAM10的本地定位；特别是通过包装成蔬菜，用于细胞外释放，贩运到质膜和内吞细胞内部化。此外，我们的目标是确定ADAM10的氨基酸基序或结构元素该会议的依赖于自噬的法规。这些策略中的每一种都是试图更好地理解 ADAM10通过自噬调节的生物学，因为该途径及其底物可以作为替代方案金黄色葡萄球菌感染和其他疾病的治疗靶标涉及内皮屏障。