Mechanisms of TOR-dependent control of autophagy in Drosophila

果蝇 TOR 依赖性自噬控制机制

• 批准号: 10459598
• 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/10459598
• 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.

项目摘要 该项目的长期目标是了解自噬的调节和生理作用，一个 蛋白质，细胞器和大量细胞质在自噬囊泡中隔离的过程 递送到溶酶体降解。这个过程在细胞中起着几种不同的至关重要的作用，作用于 回收老化或损坏的成分，以响应饥饿而提供营养来源，在某些方面 启动细胞死亡的病例。这些细胞功能是对自噬的影响的日益认识的基础 在广泛的人类疾病和正常生理过程中，例如衰老。基本的 有关自噬的问题仍有待解决，包括如何调节自噬的启动 营养和其他信号，自噬囊泡如何成熟以降解的能力以及如何 自噬的速率保持在最佳细胞存活最佳的稳态水平。令人兴奋的前景 利用自噬作为一种治疗工具，可以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遗传，生化和基于成像的方法 帮助定义完整生物体中自噬调节的机制。我们的先前 研究确定了雷帕霉素（TOR）靶标的许多关键目标和机制 途径会抑制自噬，以响应有利的营养条件。当前的提议试图 了解1）TOR信号如何与其他营养和发展线索（例如cAMP）集成 依赖性蛋白激酶A控制自噬诱导的初始步骤； 2）检验假设 描述TOR信号控制自噬体与溶酶体的融合的潜在机制 以及随后的自溶性酸化； 3）破译反馈机制 提供稳态控制，以限制自噬活动的速度和水平。 此提案中的实验因其对关键问题产生很大影响的可能性而被选择 对自噬领域很重要。该建议利用了最近开发的试剂，包括 几种自噬相关（ATG）基因的敲除，自噬活动的体内标记和新方法 细胞克隆中的遗传操作。从果蝇的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整体补充。