Mechanisms of TOR-dependent control of autophagy in Drosophila

果蝇 TOR 依赖性自噬控制机制

• 批准号: 10297218
• 负责人: Thomas P. Neufeld
• 金额: $ 32.87万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10297218
• 关键词: Address; Aging; Animals; Apoptosis; Area; Autophagocytosis; Autophagosome; Biochemical; Biological Models; Cell Culture Techniques; Cell Death; Cell Survival; Cell physiology; Cells; Clone Cells; Complement; Complex; Cues; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Cytoplasm; Defect; Development; Disease; Drops; Drosophila genus; Equilibrium; Eukaryotic Cell; Event; Fat Body; Feedback; G-Protein-Coupled Receptors; Gene Family; Genes; Genetic; Genetic Screening; Goals; Health; Human; Inflammation; Insulin Receptor; Knock-out; Link; Lysosomes; Malignant Neoplasms; Mammalian Cell; Measurement; Mediating; Mediator of activation protein; Membrane; Methods; Nerve Degeneration; Nutrient; Organ Culture Techniques; Organelles; Organism; Pathway interactions; Pattern; Phosphorylation; Physiological; Physiological Processes; Play; Process; Proteins; Proton Pump; Reagent; Recycling; Regulation; Role; Signal Transduction; Signaling Molecule; Sirolimus; Source; Starvation; Stress; System; Testing; Therapeutic; Tissues; Vesicle; Work; Yeasts; aged; base; body system; cancer cell; experimental study; extracellular; fly; genetic manipulation; imaging approach; in vivo; novel; nutrition; protein kinase A kinase; response; restoration; tool; trafficking; tumorigenesis; ubiquitin-protein ligase; vacuolar H+-ATPase

PROJECT SUMMARY The long term goal of this project is to understand the regulation and physiological roles of autophagy, a process by which proteins, organelles and bulk cytoplasm are sequestered within autophagic vesicles and delivered to the lysosome for degradation. This process plays several distinct, vital roles in the cell, acting to recycle aged or damaged components, to provide a source of nutrients in response to starvation, and in some cases to initiate cell death. These cellular functions underlie a growing appreciation for the impact of autophagy on a broad range of human illnesses and on normal physiological processes such as aging. Fundamental questions regarding autophagy remain to be addressed, including how autophagy initiation is regulated by nutrients and other signals, how autophagic vesicles mature to become competent for degradation, and how rates of autophagy are maintained at homeostatic levels optimal for cell survival. The exciting prospect of harnessing autophagy as a therapeutic tool awaits better understanding of these basic principals. The proposed studies will use genetic, biochemical and imaging-based approaches in the Drosophila system to help define mechanisms of autophagy regulation in the context of in an intact organism. Our previous studies identified a number of key targets and mechanisms through which the Target of Rapamycin (TOR) pathway inhibits autophagy in response to favorable nutrient conditions. The current proposal seeks to understand 1) how TOR signaling is integrated with other nutrient and developmental cues such as cAMP- dependent protein kinase A to control the initial steps of autophagy induction; 2) to test hypotheses that describe potential mechanisms by which TOR signaling controls the fusion of autophagosomes with lysosomes and the subsequent acidification of the autolysosome; and 3) to decipher the feedback mechanisms that provide homeostatic control limiting the rates and levels of autophagic activity. Experiments in this proposal were selected for their likelihood of having a high impact on key questions important to the field of autophagy. This proposal makes use of recently developed reagents including knockouts of several autophagy-related (Atg) genes, in vivo markers of autophagic activity, and novel methods of genetic manipulation in cell clones. Information gained from studies of autophagy in Drosophila will provide an important whole-animal complement to mammalian cell culture-based studies.

项目概要 该项目的长期目标是了解自噬的调节和生理作用，自噬是一种 蛋白质、细胞器和大量细胞质被隔离在自噬囊泡内的过程 输送至溶酶体进行降解。这个过程在细胞中发挥着几个不同的、重要的作用， 回收老化或损坏的组件，以提供应对饥饿的营养来源，并且在某些情况下 情况下启动细胞死亡。这些细胞功能是人们日益认识自噬影响的基础 广泛的人类疾病和正常的生理过程（例如衰老）。基本的 有关自噬的问题仍有待解决，包括自噬的启动如何受到调控 营养物质和其他信号，自噬囊泡如何成熟并具有降解能力，以及如何 自噬率维持在细胞存活的最佳稳态水平。令人兴奋的前景 利用自噬作为治疗工具有待更好地理解这些基本原理。 拟议的研究将在果蝇系统中使用基于遗传、生化和成像的方法 帮助定义完整有机体中自噬调节的机制。我们之前的 研究确定了雷帕霉素靶点 (TOR) 的许多关键靶点和机制 途径抑制自噬以响应有利的营养条件。目前的提案旨在 了解 1) TOR 信号如何与其他营养和发育线索（例如 cAMP-）整合 依赖蛋白激酶 A 来控制自噬诱导的初始步骤； 2）检验假设 描述 TOR 信号传导控制自噬体与溶酶体融合的潜在机制 以及随后自溶酶体的酸化； 3）破译反馈机制 提供限制自噬活动的速率和水平的稳态控制。 本提案中的实验是根据其对关键问题产生重大影响的可能性而选择的 在自噬领域具有重要意义。该提案利用了最近开发的试剂，包括 几个自噬相关 (Atg) 基因的敲除、自噬活性的体内标记以及新方法 细胞克隆中的基因操作。从果蝇自噬研究中获得的信息将提供 对基于哺乳动物细胞培养的研究的重要的全动物补充。