Phosphatidylinositol 3-Phosphate in the Regulation of Autophagic Membrane Remodeling

磷脂酰肌醇 3-磷酸在自噬膜重塑调节中的作用

基本信息

批准号：
10506784
负责人：
Ting-Sung Hsieh
金额：
$ 10万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
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：
2022-09-07 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10506784
关键词：
1-Phosphatidylinositol 3-Kinase Acute Address Advisory Committees Architecture Automobile Driving Autophagocytosis Autophagosome Award Binding Biochemical Biogenesis Biological Biological Assay Biophysics Cell physiology Cells Chemicals Coupled Data Diabetes Mellitus Diffusion Disease Disease Outcome Endoplasmic Reticulum Ensure Environment Event Functional disorder Future Generations Genetic Goals In Vitro Intracellular Membranes Lead Legionella Legionella pneumophila Link Lipids Lysosomes Malignant Neoplasms Mediating Medical center Membrane Membrane Biology Methods Modeling Molecular N-terminal Nerve Degeneration Organelles Outcome Pathologic Processes Pathway interactions Pattern Phase Phosphatidylinositols Phosphoric Monoester Hydrolases Phosphotransferases Physiological Processes Play Proteins Reaction Regulation Research Research Personnel Research Training Role Scientist Site Structure System Testing Theoretical model Therapeutic Training Tubular formation Vesicle Vps34 Phosphatidylinositol 3 Kinase biophysical techniques career career development chronic infection human disease in silico innovation live cell imaging membrane model novel therapeutic intervention optogenetics pathogen pathogenic bacteria phosphatidylinositol 3-phosphate phosphatidylinositol-3-phosphatase protein degradation quantitative imaging reconstitution recruit research and development skills spatiotemporal success tool

项目摘要

PROJECT SUMMARY/ABSTRACT Autophagy is a fundamental cellular process mediating lysosome-dependent degradation of proteins, organelles, and intracellular pathogens. Dysfunction of autophagy is associated with many diseases, including cancer, neurodegeneration, diabetes, and chronic infections. Better elucidation of the molecular mechanisms for autophagy may inspire new therapeutic approaches to these diseases. Autophagy initiation occurs at endoplasmic reticulum (ER) subdomains enriched with phosphatidylinositol 3-phosphate (PI3P). Autophagy is blocked when depleting or inactivating VPS34, the phosphatidylinositol (PI) 3-kinase responsible for PI3P synthesis during autophagy. Nevertheless, how PI3P contributes to membrane remodeling events crucial for autophagosome biogenesis remains elusive. Much is also unknown about how PI3P synthesis and turnover are orchestrated at autophagy initiation sites. Dr. Hsieh recently discovered that the Legionella PI 3-kinase MavQ generates PI3P on the ER and drives membrane remodeling. He also found that MavQ is coupled with the Legionella PI 3-phosphatase SidP to spatiotemporally modulate PI3P levels at ER subdomains, inducing vesicle/tubule budding. The striking similarity between this pathological process and autophagy initiation prompts Dr. Hsieh to dissect the molecular and physical factors that PI3P brings for autophagic membrane remodeling. Moreover, the current technical hurdles in tackling this question can mostly be cleared by using MavQ and SidP as tools. In Dr. Hsieh’s proposed research, Aim 1 will develop optogenetic methods to control PI3P generation in the cell and determine how PI3P regulates autophagic membrane remodeling using live-cell imaging and systematic genetic perturbations. Aim 2 will combine in vitro biophysical assays, optogenetic control, and quantitative imaging to elucidate how PI3P domain formation leads to membrane remodeling. Aim 3 will reconstitute a reaction-diffusion system in vitro and use this system to determine how PI 3-kinases and phosphatases drive the formation of PI3P-enriched membrane subdomains. Dr. Hsieh’s career goal is to become a leader in cell biophysics and membrane biology, focusing on physiological and pathological processes involving membrane remodeling. Training during the award period will prepare him to lead an independent research group using cell biological, biochemical, and biophysical approaches to understand the mechanistic basis of autophagic membrane remodeling. UT Southwestern Medical Center provides an excellent environment to aid Dr. Hsieh’s proposed research and career development. Dr. Hsieh has also set up an advisory committee consisting of leading scientists with complementary research expertise. Under their guidance, Dr. Hsieh will receive the necessary research training, such as preparing and using various model membrane systems, and further develop professional skills during the award period. These will significantly facilitate Dr. Hsieh's transition into an independent investigator and ensure his future success.

项目概要/摘要自噬是介导溶酶体依赖性蛋白质、细胞器、和细胞内病原体的自噬功能障碍与许多疾病有关，包括癌症，更好地阐明神经退行性变、糖尿病和慢性感染的分子机制。自噬可能会激发这些疾病的新治疗方法。富含磷脂酰肌醇 3-磷酸 (PI3P) 的内质网 (ER) 子结构域是自噬。当耗尽或灭活 VPS34（负责 PI3P 的磷脂酰肌醇 (PI) 3-激酶）时，会被阻断然而，PI3P 如何促进膜重塑事件对于细胞膜重塑至关重要。自噬体的生物发生仍然难以捉摸，关于 PI3P 的合成和周转如何也还不清楚。 Hsieh 博士最近发现军团菌 PI 3 激酶 MavQ 他还发现 MavQ 与 ER 上产生 PI3P 并驱动膜重塑。军团菌 PI 3-磷酸酶 SidP 时空调节 ER 子域的 PI3P 水平，诱导这种病理过程与自噬起始之间惊人的相似。促使谢博士剖析PI3P为自噬膜带来的分子和物理因素此外，目前解决这个问题的技术障碍大部分可以通过使用来解决。在Hsieh博士提出的研究中，Aim 1将开发光遗传学方法来控制。细胞中 PI3P 的生成并利用活细胞确定 PI3P 如何调节自噬膜重塑目标 2 将结合体外生物物理测定和光遗传学测定。控制和定量成像来阐明 PI3P 结构域的形成如何导致膜重塑。 3 将在体外重建反应扩散系统，并使用该系统来确定 PI 3-激酶和磷酸酶驱动富含 PI3P 的膜子域的形成。谢博士的职业目标是成为。细胞生物物理学和膜生物学领域的领导者，专注于生理和病理过程获奖期间涉及膜重塑的培训将为他领导独立做好准备。研究小组使用细胞生物学、生物化学和生物物理方法来了解其机制 UT西南医学中心为自噬膜重塑的基础提供了良好的环境。为了帮助谢博士提出的研究和职业发展，谢博士还成立了一个咨询委员会。谢博士将由具有互补研究专业知识的顶尖科学家组成。接受必要的研究培训，例如准备和使用各种模型膜系统，以及在获奖期间进一步发展专业技能，这将极大地促进谢博士的发展。转变为一名独立调查员并确保他未来的成功。