Control of vesicular trafficking in the hepatocyte.

控制肝细胞中的囊泡运输。

• 批准号: 8838778
• 负责人: ANA MARIA CUERVO
• 金额: $ 30万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8838778
• 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; receptor mediated endocytosis; reconstitution; response; tool; trafficking

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）：内吞作用和自噬是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。我们的研究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隔室；2)两者都有助于细胞蛋白水解；3)这两种途径都需要调节囊泡的运输，这些囊泡以动态的方式相互作用，在其成熟过程中经历裂变和融合。基于先前的研究结果，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，并参与肝脏疾病的发病机制。本建议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)从机制上定义囊泡相关蛋白