Organotypic Slice Culture Model of CHIP Mediated Neuroprotection

CHIP 介导的神经保护的器官型切片培养模型

• 批准号: 10647093
• 负责人: Jonathan C. Schisler
• 金额: $ 2.93万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10647093
• 关键词: 5' -AMP-activated protein kinase; Acceleration; Active Sites; Adult; Aging; Alzheimer' s Disease; Apoptosis; Atrophic; Binding Proteins; Biochemical; Biology; Cardiovascular Diseases; Cell Culture Techniques; Cell Death; Cell model; Cell physiology; Cells; Cellular Metabolic Process; Cellular Stress; Cerebellar Ataxia; Cerebellar degeneration; Cerebellum; Code; Cognition; Complex; Defect; Degenerative Disorder; Disease; Energy Metabolism; Enzymes; Functional disorder; Genetic Diseases; Goals; Health; Heart failure; Heat-Shock Response; Human Genetics; Hypogonadism; Impaired cognition; Inflammatory; Link; Mediating; Mediator; Metabolic; Metabolic stress; Metabolism; Modeling; Molecular; Molecular Chaperones; Molecular Conformation; Motor; Movement; Mus; Mutation; Nerve Degeneration; Neurologic; Neurons; Pathology; Pathway interactions; Patients; Phenotype; Phosphotransferases; Physiological; Play; Pre-Clinical Model; Predisposition; Protein Kinase Interaction; Proteins; Proteolytic Processing; Purkinje Cells; Quality Control; RIPK1 gene; Reaction; Regulation; Role; Signal Pathway; Signal Transduction; Slice; Spinocerebellar Ataxias; Stress; System; Testing; Tissues; Ubiquitin; age related; autosome; biophysical techniques; body system; clinical phenotype; disease-causing mutation; druggable target; engineered stem cells; flexibility; human disease; induced pluripotent stem cell; loss of function; member; mouse model; multicatalytic endopeptidase complex; neuroprotection; novel; novel therapeutics; parent grant; pharmacologic; pre-clinical; precision medicine; protein degradation; protein folding; protein function; proteostasis; response; selective expression; sensor; small molecule; stressor; ubiquitin ligase

ABSTRACT – PARENT GRANT Protein quality control is vital for cellular function, encompassing pathways that help proteins fold and maintain structural conformations, as well as cellular systems that ultimately degrade proteins. Several degenerative diseases, including those that are accompanied by abnormal aging, are associated with altered protein quality control, highlighting the importance of protein fidelity in both health and disease. CHIP (carboxyl terminus of heat shock 70-interacting protein) is a multi-functional enzyme with distinct activities that contribute both to protein folding and protein degradation. Our lab recently identified the first human disease associated with CHIP mutations, with phenotypes that include accelerated aging, cerebellar ataxia and degeneration, cognitive dysfunction, and hypogonadism. Two known CHIP substrates, receptor-interacting protein kinase 3 (RIPK3) and AMP-activated kinase (AMPK), are regulated by either the degradative or re-folding activities of CHIP, respectively. These kinases regulate important cellular processes, necroptosis (an inflammatory form of programmed cell death) and cellular metabolism, both of which are central to the ability of cells to react to the stress that occurs with aging or in pathophysiological conditions. In this proposal, we seek to define the role of protein quality control in aging by determining how the various activities of CHIP regulate these recently identified substrates, and the mechanism that drives the sensitivity to cell stress when CHIP function is compromised. Our approach includes novel pre-clinical models of accelerated aging, cutting-edge cell models, and determining the molecular movements of CHIP when engaged with its substrate. We then ask how disease-causing mutations in CHIP mechanistically drive the molecular and cellular phenotypes associated with CHIP dysfunction. Finally, we will use our preclinical models to determine how aging and disease pathologies are altered when necroptosis is inhibited pharmacologically, or when CHIP function is restored genetically. The ultimate goal of these studies is to identify druggable targets of CHIP-regulated signaling pathways that impact age-dependent progression of degenerative conditions.

摘要 – 家长补助金 蛋白质质量控​​制对于细胞功能至关重要，包括帮助蛋白质折叠和维持的途径 结构构象，以及最终降解蛋白质的细胞系统。几处退化 疾病，包括那些伴有异常衰老的疾病，与蛋白质质量改变有关 控制，强调蛋白质保真度在健康和疾病中的重要性。 CHIP（羧基末端 热休克 70 相互作用蛋白）是一种多功能酶，具有独特的活性，有助于 蛋白质折叠和蛋白质降解。我们的实验室最近发现了第一种与以下疾病相关的人类疾病 CHIP 突变，其表型包括加速衰老、小脑共济失调和退化、认知 功能障碍和性腺功能减退症。两种已知的 CHIP 底物，受体相互作用蛋白激酶 3 (RIPK3) 和 AMP 激活激酶 (AMPK)，受 CHIP 的降解或重折叠活性调节， 分别。这些激酶调节重要的细胞过程，坏死性凋亡（坏死性凋亡的一种炎症形式） 程序性细胞死亡）和细胞代谢，两者都是细胞对细胞反应能力的核心 随着衰老或病理生理条件而发生的压力。在本提案中，我们寻求定义角色 通过确定 CHIP 的各种活性如何调节衰老过程中的蛋白质质量控​​制 最近发现的底物，以及 CHIP 时驱动细胞应激敏感性的机制 功能受到损害。我们的方法包括加速衰老的新型临床前模型、尖端的 细胞模型，并确定 CHIP 与其底物接触时的分子运动。然后我们问 CHIP 中的致病突变如何机械地驱动分子和细胞表型 与 CHIP 功能障碍有关。最后，我们将使用我们的临床前模型来确定衰老和 当程序性坏死在药理学上被抑制时，或者当 CHIP 功能被抑制时，疾病病理会发生改变。 基因恢复。这些研究的最终目标是确定 CHIP 调节的药物靶标 影响退行性疾病的年龄依赖性进展的信号通路。