Control of vesicular trafficking in the hepatocyte.

控制肝细胞中的囊泡运输。

基本信息

批准号：
8479622
负责人：
ANA MARIA CUERVO
金额：
$ 71.99万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8479622
关键词：
Affect Animals Autophagocytosis Biochemical Biological Assay Cell Line Cells Cellular biology Cholesterol Communicable Diseases Complex Cultured Cells Development Diabetes Mellitus Disease Endocytic Vesicle Endocytosis Endocytosis Pathway Endosomes Event Failure Fluorescence Microscopy Functional disorder Goals Guanosine Triphosphate Phosphohydrolases Health Heat-Shock Response Hepatitis Hepatocyte Homeostasis Hybrids Immune System Diseases In Vitro Kinetics Knowledge Lipids Liver Liver diseases Lysosomes Malignant Neoplasms Mediating Membrane Membrane Lipids Metabolic Diseases Microtubules Molecular Molecular Chaperones Motor Mus Names Obesity Oxidative Stress Pathogenesis Pathology Pathway interactions Physiological Population Procedures Process Protein C Inhibitor Proteins Proteolysis Proteomics Rattus Recruitment Activity Relative (related person)Starvation Stimulus System Technology Testing Therapeutic Vacuole Vesicle base cell motility cell type chemical genetics functional decline genetic manipulation human disease knock-down late endosome liver function new therapeutic target novel protein complex protein degradation public health relevance receptor mediated endocytosis reconstitution response tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Endocytosis and autophagy represent fundamental processes that are essential to the health of many cell types, including hepatocytes. Previous studies from our groups and others have identified cross-talk between endocytosis and autophagy: 1) specific compartments are shared between them; 2) both contribute to cellular proteolysis and 3) both pathways require regulated trafficking of vesicles that interact in a dynamic manner, undergoing fission and fusion as part of their maturation process. Based on prior findings, we propose that dysfunction of the endocytic or autophagic trafficking pathways can have deleterious effects on normal liver function and contribute to the pathogenesis of hepatic disorders. The overall goal of this proposal is to characterize at the molecular level the functional interaction between the endocytic and the autophagic pathways and their contribution to cellular homeostasis. To this end we will: 1) mechanistically define vesicle- associated protein complexes that regulate motor recruitment and activity required for endocytic/autophagic vesicle processing by fission and fusion; 2) determine the consequences of changes in membrane lipid composition on the interactions between endocytic and autophagic pathways and 3) characterize quantitatively the functional interplay between endocytosis and autophagy. We will use chemical and genetic manipulations in cultured hepatocyte cell lines and in rat and mouse liver combined with biochemical and morphological approaches to answer these questions. The complementary expertise of the two co-PIs, Dr. Allan Wolkoff (endocytosis and liver pathophysiology) and Dr. Ana Maria Cuervo (autophagy and cell biology) greatly enhances their ability to discover interactions between the endocytic and the autophagic pathways in liver and to study the consequences that failure in one of these pathways has on the other. The major significance of the proposed studies resides in the fact that alterations in endocytosis and autophagy have been observed in human diseases that include protein conformational disorders (e.g. alpha1-antitrypsin deficiency), cancer, metabolic disorders (e.g. diabetes, obesity) and infectious and immune diseases. However, incomplete knowledge regarding details of endocytosis and autophagy limits development of mechanism-based therapeutics. Successful completion of these studies will ultimately provide the tools needed to identify novel therapeutic targets to restore normal liver function or perhaps slow the functional decline associated with common liver disorders resulting from dysfunction of the endosome/lysosome system.

描述（由申请人提供）：内吞作用和自噬代表了对包括肝细胞在内的许多细胞类型的健康至关重要的基本过程。我们小组和其他人之前的研究已经确定了内吞作用和自噬之间的串扰：1）它们之间共享特定的区室； 2) 两者都有助于细胞蛋白水解，3) 两种途径都需要以动态方式相互作用的囊泡的受调节运输，在其成熟过程中经历裂变和融合。根据先前的发现，我们提出内吞或自噬运输途径的功能障碍可能对正常肝功能产生有害影响，并导致肝脏疾病的发病机制。该提案的总体目标是在分子水平上表征内吞和自噬途径之间的功能相互作用及其对细胞稳态的贡献。为此，我们将：1）机械地定义囊泡相关蛋白通过裂变和融合调节内吞/自噬囊泡加工所需的运动募集和活动的复合物； 2) 确定膜脂成分变化对内吞和自噬途径之间相互作用的影响；3) 定量表征内吞作用和自噬之间的功能相互作用。我们将在培养的肝细胞系以及大鼠和小鼠肝脏中使用化学和遗传操作，并结合生化和形态学方法来回答这些问题。两位共同首席研究员 Allan Wolkoff 博士（内吞作用和肝脏病理生理学）和 Ana Maria Cuervo 博士（自噬和细胞生物学）的互补专业知识极大地增强了他们发现肝脏内吞和自噬途径之间相互作用的能力，并研究这些途径之一失败对另一途径造成的后果。拟议研究的主要意义在于，在人类疾病中观察到内吞作用和自噬的改变，包括蛋白质构象紊乱（例如α1-抗胰蛋白酶缺乏症）、癌症、代谢性疾病（例如糖尿病、肥胖）以及感染性和免疫性疾病疾病。然而，关于内吞作用和自噬细节的不完整知识限制了基于机制的治疗的发展。这些研究的成功完成将最终提供确定新的治疗靶点所需的工具，以恢复正常的肝功能，或者可能减缓与内体/溶酶体系统功能障碍导致的常见肝脏疾病相关的功能衰退。