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

• 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
• 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

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 S.金黄色葡萄球菌是社区和医疗机构中大量感染的原因。特别是作为 抗生素耐药菌株的数量和发生率继续上升，需要替代干预措施 方法变得越来越重要。开发新疗法的一个策略是识别和阻断宿主 病原体利用的致病途径。自噬就是这样一种途径。自噬是一种 一种保守的、普遍存在的细胞过程，其中双膜自噬体吞噬受损的细胞质 成分，并将其作为溶酶体降解的目标;然而，最近的研究表明， 在病原体耐受性和清除中的作用。 卡德韦尔和托雷斯实验室合作进行的实验表明， 对于介导自噬体形成的蛋白质ATG16L1，在S.金黄色葡萄球菌耐受性使用体内模型 自噬丧失的情况下，ATG16L1几乎完全消除，增加的易感性致死 发现挑战依赖于S.金黄色葡萄球菌毒力因子、金黄色葡萄球菌毒素。经过进一步 在实验中，显示缺乏ATG16L1的内皮细胞显示更高水平的血浆 膜脂毒素受体ADAM10。这些数据表明，自噬在细胞凋亡中起着负调节作用。 ADAM 10.然而，目前尚不清楚自噬是如何调节细胞质降解途径， 膜结合型ADAM10 我们的目的是确定宿主因素，机制和/或途径，差异调节， 自噬影响ADAM 10水平。使用我们为此目的开发的体外系统，我们最初的 数据表明ATG16L1独立于溶酶体或蛋白酶体调节ADAM 10。相反，细胞 缺乏ATG16L1的小鼠显示含有ADAM 10的细胞外囊泡的产生减少。我们计划继续 精确测试ATG16L1和自噬如何影响ADAM 10的天然定位;特别是通过 包装成囊泡，用于细胞外释放，运输到质膜，和内吞 内化此外，我们的目标是确定ADAM 10的氨基酸基序或结构元件， 赋予其自噬依赖性调节。这些策略中的每一个都是为了更好地理解 自噬调节ADAM10的生物学，因为这一途径及其底物可以作为替代 治疗S.金黄色葡萄球菌感染和其他涉及内皮屏障的疾病。