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）：内吞作用和自噬代表了对许多细胞类型（包括肝细胞）的健康至关重要的基本过程。我们小组和其他人以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和3）两种途径都需要以动态方式相互作用的囊泡的调节运输，经历裂变和融合作为其成熟过程的一部分。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，并有助于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内稳态的贡献。为此，我们将：1）机械定义囊泡相关蛋白 复合物，调节运动募集和活动所需的内吞/自噬囊泡加工的分裂和融合; 2）确定的后果，在膜脂质组成的变化之间的相互作用的内吞和自噬途径和3）定量的特点之间的功能相互作用的内吞和自噬。我们将在培养的肝细胞系和大鼠和小鼠肝脏中使用化学和遗传操作，结合生物化学和形态学方法来回答这些问题。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）的互补专业知识大大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个的后果。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，这些疾病包括蛋白质构象紊乱（例如α 1-抗胰蛋白酶缺乏症）、癌症、代谢紊乱（例如糖尿病、肥胖症）以及感染性和免疫性疾病。然而，关于内吞作用和自噬的细节的不完整知识限制了基于机制的疗法的发展。这些研究的成功完成将最终提供所需的工具，以确定新的治疗靶点，以恢复正常的肝功能或可能减缓与内体/溶酶体系统功能障碍引起的常见肝脏疾病相关的功能下降。