Functional Dissection of Autophagosome Biogenesis

自噬体生物发生的功能剖析

基本信息

批准号：
8868838
负责人：
Liang Ge
金额：
$ 8.64万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-01 至 2017-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8868838
关键词：
1-Phosphatidylinositol 3-Kinase Accounting Address Aging Attention Autophagocytosis Autophagosome Award Biochemical Biochemical Reaction Biochemistry Biogenesis Biological Assay Cell Survival Cell physiology Cells Complement Complex Coupled Development Digestion Disease Dissection Educational process of instructing Electron Microscopy Event Fibrinogen Fractionation Functional disorder Generations Genetic Goals Golgi Apparatus Homeostasis Human Hypoxia Image Imaging Techniques Immune System Diseases Infection Knowledge Lead Learning Location Maintenance Malignant Neoplasms Mass Spectrum Analysis Membrane Membrane Proteins Mentors Molecular Morphology Myopathy Nature Nerve Degeneration Organelles Pathogenesis Phase Physiological Process Protein-Serine-Threonine Kinases Proteins Quality Control Reaction Research Research Personnel Research Training Resolution Role Scientist Set protein Signal Transduction Source Staging Starvation Stress Techniques Therapeutic Training Ubiquitin Vesicle Writing base cellular imaging crosslink experience genetic manipulation macromolecule novel pathogen phosphatidylinositol 3-phosphate public health relevance quantitative imaging reconstitution spatiotemporal success therapeutic development treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Autophagy is a fundamental mechanism for maintenance of cellular homeostasis through self-digestion. Abnormal autophagy is closely related to many human disorders such as cancer, aging, and neurodegeneration. A deep mechanistic understanding of autophagy is crucial for the development of therapeutic strategies against these diseases. Autophagosome biogenesis is intimately associated with the capture and destruction of macromolecules whose turnover is executed in the autophagic process. It entails a spatiotemporal orchestration of protein-membrane interactions which is not fully understood. Dr. Ge has established a functional assay based on cell-free reconstitution of autophagosome biogenesis, through which he has made a further step towards the understanding of autophagosome biogenesis by identifying the ER-Golgi intermediate compartment as a key membrane source of the autophagosome. In this study, Dr. Ge seeks to extend this functional assay to decipher the underlying molecular actions generating the autophagosome. In Aim 1, Dr. Ge will establish a set of new cell-free assays together with cell-imaging and genetics to investigate the initial step of autophagosome biogenesis by focusing on an autophagic signal-induced membrane mobilization event generating the autophagic membrane precursor. In Aim 2, Dr. Ge will develop a systematic protein fractionation approach to identify novel protein factors in autophagosome biogenesis, as well as to define the functional role of each factor in the protein-membrane network of autophagy. Dr. Ge's long term goal is to understand the molecular nature of autophagy in physiological and pathological settings for the purpose of autophagy-modulating therapy. Training in the mentored phase will prepare Dr. Ge to lead an independent research team using biochemical reconstitution, cell imaging, genetic manipulation, and mass spectrometry to address the fundamental questions of autophagy, as well as to understand the pathogenesis of autophagy-related diseases. Training under this award will include: learning new techniques, such as super-resolution imaging, electron microscopy and quantitative mass spectrometry, acquiring more experience in biochemical reconstitution and fractionation, and expanding knowledge and expertise in autophagy, as well as teaching and writing. Completion of the research and training will greatly facilitate Dr. Ge's transition and success as an independent investigator.

描述（由申请人提供）：自噬是通过自我消化维持细胞稳态的基本机制，异常自噬与许多人类疾病密切相关，例如癌症、衰老和神经退行性变。针对这些疾病的治疗策略的发展与自噬过程中执行的大分子的捕获和破坏密切相关。葛博士建立了一种基于自噬体生物发生的无细胞重建的功能测定，通过识别 ER- ，他在理解自噬体生物发生方面又迈出了一步。高尔基体中间室是自噬体的关键膜源。在这项研究中，葛博士试图扩展这种功能测定，以破译产生自噬体的潜在分子作用。在目标 2 中，葛博士将建立一套新的无细胞实验，并结合细胞成像和遗传分析，通过关注自噬信号诱导的膜动员事件来研究自噬体生物发生的初始步骤，从而产生自噬膜前体。葛博士将开发一种系统的蛋白质分级分离方法，以鉴定自噬体生物发生中的新蛋白质因子，并确定每个因子在自噬蛋白质膜网络中的功能作用。葛博士的长期目标是了解生理和病理环境中自噬的分子性质，以实现自噬调节治疗。指导阶段的培训将使葛博士能够领导一个使用生化重建、细胞成像、遗传学的独立研究团队。该奖项的培训将包括：学习新技术，例如超分辨率成像、电子技术。显微镜和定量质谱分析，获得更多生化重建和分离方面的经验，扩大自噬方面的知识和专业知识，以及教学和写作的完成将极大地促进葛博士作为一名独立研究者的转变和成功。