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.

项目概要 金黄色葡萄球菌是社区和医疗机构中大量感染的罪魁祸首。 抗生素耐药菌株的数量和发生率持续上升，需要替代干预 方法变得越来越重要，开发新疗法的策略是识别和阻断宿主。 自噬就是这样一种途径。 保守的、普遍存在的细胞过程，其中双膜自噬体吞噬受损的胞质 然而，最近的研究证明了其关键性 在病原体耐受和清除中的作用。 卡德威尔和托雷斯实验室合作进行的实验证明了其至关重要的作用 ATG16L1，一种介导自噬体形成的蛋白质，在金黄色葡萄球菌耐受中使用体内模型。 当 ATG16L1 几乎完全被废除时，自噬丧失，致死的易感性增加 进一步发现，挑战取决于金黄色葡萄球菌毒力因子α-毒素的产生。 实验表明，缺乏 ATG16L1 的内皮细胞表现出更高水平的血浆 这些数据表明自噬对膜α毒素受体ADAM10起着负调节作用。 然而，自噬（一种细胞质降解途径）如何调节其水平仍不清楚。 膜结合 ADAM10。 我们的目标是确定受不同调节的宿主因素、机制和/或途径 使用我们为此目的开发的体外系统，我们最初研究了影响 ADAM10 水平的自噬。 数据表明 ATG16L1 独立于溶酶体或蛋白酶体调节 ADAM10。 缺乏 ATG16L1 显示含有 ADAM10 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 ADAM10 的天然定位； 包装成囊泡，用于细胞外释放、运输到质膜和内吞 此外，我们的目标是确定 ADAM10 的氨基酸基序或结构元件。 这些策略中的每一种都是为了更好地理解其自噬依赖性调节。 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途径 治疗金黄色葡萄球菌感染和涉及内皮屏障的其他病症的目标。