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

基本信息

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
项目类别：
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&apos -AMP-activated protein kinase Acceleration Active Sites Adult Aging Alzheimer&apos 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

项目摘要

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 - 父母赠款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并维持的途径结构构象以及最终降解蛋白质的细胞系统。几个退化疾病，包括伴有异常衰老的疾病，与蛋白质质量改变有关控制，强调了蛋白质保真在健康和疾病中的重要性。芯片（羧基末端热休克70相互作用蛋白）是一种多功能酶，具有不同的活性，这两种酶有助于蛋白质折叠和蛋白质降解。我们的实验室最近确定了与芯片突变，具有包括加速衰老的表型，小脑共济失调和变性，认知功能障碍和性能障碍。两个已知的芯片底物，受体相互作用蛋白激酶3（RIPK3）和AMP激活的激酶（AMPK）受芯片的降解或重新折叠活性的调节，这些激酶调节重要的细胞过程，坏死性（一种炎症形式程序性细胞死亡）和细胞代谢，这两者都是细胞反应能力的核心在衰老或病理生理状况下发生的压力。在此提案中，我们试图定义角色通过确定芯片的各种活动如何调节蛋白质质量控制的蛋白质质量控制最近确定的底物以及芯片时驱动细胞应激敏感性的机制功能受到损害。我们的方法包括新型的加速衰老，尖端的临床前模型细胞模型，并确定芯片与底物互动时的分子运动。然后我们问芯片中引起疾病的突变如何机械驱动分子和细胞表型与芯片功能障碍相关。最后，我们将使用临床前模型来确定衰老和当坏死性抑制药物或芯片功能为时，疾病病理会改变一般恢复。这些研究的最终目的是确定CHIP调节的可吸毒目标影响退化条件的年龄依赖性进展的信号通路。