TDP43 Degradation by the Lysosomal Proteases in Amyotrophic Lateral Sclerosis

肌萎缩侧索硬化症中溶酶体蛋白酶对 TDP43 的降解

• 批准号: 10524706
• 负责人: Paul Joseph Sampognaro
• 金额: $ 19.05万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10524706
• 关键词: Alkalinization; Amyotrophic Lateral Sclerosis; Area; Autophagocytosis; Basic Science; Biological Assay; Cathepsins; Cathepsins B; Cell Compartmentation; Cell Survival; Cell physiology; Cells; Clinical; Clinical Trials; Complex; Critical Pathways; Data; Development; Disease; Enzymes; Faculty; Fellowship; Foundations; Functional disorder; Funding; Goals; Half-Life; Homeostasis; Human; Impairment; In Vitro; Incubated; Induced pluripotent stem cell derived neurons; Investigation; K-Series Research Career Programs; Knowledge; Lead; Lysosomes; Mass Spectrum Analysis; Measures; Membrane; Mentors; Mentorship; Methods; Modeling; Morphology; Motor; Motor Neurons; Mutation; Nerve Degeneration; Neurobiology; Neurodegenerative Disorders; Neurology; Neurons; Organelles; Pathogenesis; Pathogenicity; Pattern; Peptide Hydrolases; Phase; Pilot Projects; Process; Proteins; Recycling; Research; Research Personnel; Residencies; Resistance; Running; Scientist; Site; Solid; Testing; Therapeutic; Time; Training; Work; age related; biological adaptation to stress; career; design; experience; gain of function; induced pluripotent stem cell; knock-down; loss of function; medical schools; motor deficit; motor neuron degeneration; mutant; neuron loss; novel; novel therapeutics; overexpression; preservation; prevent; protein TDP-43; protein aggregation; purge; skills; undergraduate student

PROJECT SUMMARY/ABSTRACT Amyotrophic lateral sclerosis (ALS) is a progressive, uniformly fatal neurodegenerative disorder caused by the loss of upper and lower motor neurons. Despite multiple advanced clinical trials, there are currently no therapies that can stabilize or reverse the motor deficits of ALS. My goal in pursuing a K08 Mentored Clinical Scientist Research Career Development Award is to acquire the knowledge and practical training to make major advances in our understanding of the mechanisms underlying neuronal loss in ALS. With my career, I seek to develop novel therapies that will prevent motor neuron death and preserve function. In 2006 Neumann and colleagues discovered that TAR DNA-binding protein 43 (TDP43) accumulated abnormally within diseased neurons and represented a unifying, end-stage neuropathologic hallmark of ALS. With this observation, the field then converged on the cellular process of autophagy — the method by which cells use lysosomes to purge proteins aggregates and maintain homeostasis — as a critical pathway involved in ALS’ pathogenesis. Lysosomes, however, are tremendously intricate and not well understood compartments within neurons. Lysosomes house many complex, hydrolytic enzymes; they assist in adaptive stress responses; and they promote cell survival broadly. While it has been shown that TDP43 can be directed to the lysosome for clearance, past efforts have stopped their investigations at the lysosomal membrane, presuming that presuming that TDP43 degradation happens efficiently by an array of unspecified enzymes thereafter. My preliminary data demonstrates that only a subset of lysosomal enzymes (cathepsins B, D, E, G, L, K, S, and V) can degrade TDP43 in a pH-dependent manner and that ALS-causing TDP43 mutations have the potential to disrupt the functions of these enzymes. Therefore, I hypothesize that cathepsins B, D, E, G, L, K, S, and V are responsible for TDP43 degradation, pathogenic TDP43 mutations are capable of conferring resistance to these cathepsins, and that impaired cathepsin activity (whether due to TDP43 mutations or age- related changes in pH) contributes to TDP43 buildup, TDP43 aggregation, and neurodegeneration over time. This proposal builds upon my solid foundation in neurobiology and neurology that I have cultivated working as a basic research scientist during my undergraduate, medical school, residency, fellowship, and early faculty years. My current research mentor has an established record of impactful discoveries in the field of neurodegeneration. I have also assembled a team of highly accomplished advisors at UCSF to guide me through this next phase of training on my path to becoming an independent investigator. My training plan is specifically designed to provide me with the mentorship, training in advanced experimental skills, and experience required to run a research group. Completing the research and obtaining the skill sets and mentorship outlined in this proposal will prepare me well to obtain R01 or equivalent funding to begin my career as an independent investigator.

项目摘要/摘要 肌萎缩性侧索硬化症（ALS）是一种进行性，统一致命的神经退行性疾病 上下运动神经元的丧失。尽管进行了多次高级临床试验，但目前却没有 可以稳定或逆转电动机定义ALS的疗法。我追求K08指导临床的目标 科学家研究职业发展奖是要获得知识和实用培训以使 我们对ALS神经元损失基本机制的理解的重大进展。我的职业生涯，我 寻求开发新的疗法，以防止运动神经元死亡并保留功能。 2006年，诺伊曼（Neumann）及其同事发现焦油DNA结合蛋白43（TDP43）积累 交替地在解剖的神经元内代表ALS的统一终端神经病理学标志。 通过此观察，该领域随后在自噬的细胞过程中转换为 细胞使用溶酶体清除蛋白质聚集体并保持体内平衡 - 作为涉及的关键途径 在ALS的发病机理中。然而，溶酶体非常复杂，隔离室尚不清楚 在神经元内。溶酶体容纳许多复杂的水解酶；他们有助于适应性压力 回应；它们广泛促进细胞生存。虽然已经证明TDP43可以针对 溶酶体清除，过去的努力停止了他们在溶酶体膜的调查，假设 假定TDP43降解通过一系列未指定的酶有效地发生。 我的初步数据表明，只有一部分溶酶体酶（组织蛋白酶B，D，E，G，L，K，S，S， v）可以以pH的方式降解TDP43，并且引起ALS的TDP43突变具有 破坏这些酶功能的潜力。因此，我假设组织蛋白酶B，d，e，g，l，k， S和V负责TDP43降解，致病性TDP43突变能够赋予 对这些组织蛋白酶的耐药性，并且降低了组织蛋白酶活性（无论是由于TDP43突变还是年龄引起的 随着时间的流逝，pH）的相关变化促进了TDP43的堆积，TDP43聚集和神经变性。 这项建议是基于我在神经生物学和神经病学上的坚实基础，我已经培养了 我的本科，医学院，住所，奖学金和早期教师期间的基础研究科学家 年。我目前的研究导师在该领域有着有影响力的发现的既定记录 神经变性。我还在UCSF组建了一支高度成就的顾问团队，以指导我 通过下一阶段的培训，我成为独立研究者的道路。我的培训计划是 专门为我提供指导，高级实验技能的培训以及 经营研究小组所需的经验。完成研究并获得技能集和 该提案中概述的资术将为我做好准备，以便获得R01或同等资金开始我的 作为独立调查员的职业。