Modulating Alzheimer’s Disease by mTORC1 inhibition to augment lysosomal activity

通过抑制 mTORC1 增强溶酶体活性来调节阿尔茨海默病

基本信息

批准号：
10705289
负责人：
Travis Lear
金额：
$ 12.25万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10705289
关键词：
Affect Age Aging Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease related dementia Animal Model Autophagocytosis Award Biogenesis Biological Biology Biology of Aging Brain Caregivers Cell Culture Techniques Cell Line Cell model Cells Central Nervous System Collection Communities Complex Data Environment FRAP1 gene Future Generations Genes Genetic Goals Human Impairment In Vitro Incidence Inflammation Intervention Knockout Mice Knowledge Lead Leadership Lesion Lysosomes MAPT gene Mentors Mentorship Metabolism Modeling Molecular Biology Mus Nerve Degeneration Neurobiology Neurodegenerative Disorders Neurons Pathogenesis Pathogenicity Pathway interactions Patients Phenotype Prevalence Process Proteins Research Research Personnel Risk Factors Role Route Sirolimus Stem Cell Development Structure System Techniques Testing Therapeutic Time Toxic effect Training Ubiquitination Universities Up-Regulation Work aged brain tissue career career development cognitive function demographics detection of nutrient experience experimental study improved in vivo induced pluripotent stem cell inhibitor insight mouse model novel novel therapeutics pharmacologic pleiotropism prevent protein aggregation protein degradation screening skills small molecule stem cell model symposium tau Proteins tau aggregation tau mutation therapeutically effective tissue culture tool ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY/ABSTRACT: This proposal is in application for a Pathway to Independence Award for Dr. Travis Lear at the Aging Institute at the University of Pittsburgh. Dr. Lear has extensive experience in the molecular biology of ubiquitination in aging, inflammation, autophagy, and lysosome biology. This K99/R00 would be a crucial step for Dr. Lear as part of his career goal of reaching research independence. The focus of Dr. Lear’s future lab will be to study the mechanisms of protein degradation and lysosomal activity in neurodegeneration in pursuit of uncovering new therapeutic avenues. Under this K99/R00 award, Dr. Lear would have protected time for additional training in neurobiology techniques and aging biology, and for career development to facilitate a successful transition to research independence. This mentorship team of experts in aging, lysosomal, and neurobiology combined with the scientific environment at the Aging Institute will be ideal for Dr. Lear’s training. The scientific expertise training will entail 1) in vitro primary tissue culture skill, 2) study of iPSC development, differentiation, and gene-editing, 3) generation and phenotypic characterization of mouse models of neurodegeneration. Dr. Lear’s leadership training will focus on 1) integration to scientific community through networking, 2) enhancing his mentoring skills, 3) improving grantsmanship. Successful completion of the proposed research and training plan will provide the knowledge and experience necessary to progress toward Dr. Lear’s career goal of becoming an independent investigator studying neuronal aging and Alzheimer’s Disease (AD). To accomplish this, Dr. Lear proposes to study a new mechanism of proteolytic control of lysosomal activation to augment processing of pathogenic tau protein aggregates in models of AD. Specifically, Dr. Lear has elaborated a model in which a key mTORC1 inhibitor protein, KPTN, is potently controlled by the E3 ubiquitin ligase PDZRN3, which ubiquitinates and fates KPTN for degradation. Also, unbiased screening yielded a small molecule KPTN activator which increases KPTN protein level, inhibits mTORC1 activity, and increases lysosomal number and activity. Excitingly, pharmacological augmentation of KPTN reduces tau protein aggregation in vitro, which has therapeutic implications for the treatment of Alzheimer’s Disease. The net effect of KPTN augmentation would therefore aid in clearance of toxic protein aggregates by activation of autophagy. This leads to the central hypothesis that PDZRN3 control of KPTN affects neuronal lysosomal activity and that genetic or pharmacological activation of KPTN may be an avenue to reduce tau protein aggregates. Two aims will interrogate this hypothesis: (1) To examine the mechanism and biologic effect of the PDZRN3-KPTN axis on tau-aggregation in vitro with inducible pluripotent stem cell (iPSC) and primary cell models, and (2) to examine this mechanism and effect using genetic and pharmacological approaches in animal models of tau spreading. Dr. Lear will also pursue a structured training plan including formal course work, conferences, and hands-on training of new techniques, under the guidance of primary mentor Dr. Toren Finkel, and co-mentors Dr. Stacey Rizzo and Dr. Bill Chen.

项目概要/摘要：该提案正在为老龄化研究所的 Travis Lear 博士申请独立之路奖。匹兹堡大学的李尔博士在衰老泛素化的分子生物学方面拥有丰富的经验，作为李尔博士研究的一部分，K99/R00 将是关键的一步。实现研究独立性的职业目标。李尔博士未来实验室的重点将是研究神经变性中蛋白质降解和溶酶体活性的机制，以寻求新的发现根据这个 K99/R00 奖项，李尔博士将有时间接受额外的培训。神经生物学技术和衰老生物学，以及职业发展以促进成功过渡该导师团队由衰老、溶酶体和神经生物学领域的专家组成。老龄研究所的科学环境非常适合李尔博士的科学专业知识培训。将需要 1) 体外原代组织培养技能，2) iPSC 发育、分化和基因编辑的研究， 3）李尔博士领导的小鼠神经变性模型的生成和表型表征。培训将侧重于 1) 通过网络融入科学界，2) 提高指导技能， 3) 改进资助方式。成功完成拟议的研究和培训计划将提供实现李尔博士成为独立人士的职业目标所需的知识和经验研究神经衰老和阿尔茨海默病 (AD) 的研究人员为了实现这一目标，李尔博士建议：研究溶酶体激活的蛋白水解控制新机制，以增强致病性 tau 蛋白的加工具体来说，Lear 博士详细阐述了一个模型，其中关键的 mTORC1 存在。抑制剂蛋白 KPTN 受 E3 泛素连接酶 PDZRN3 有效控制，该酶可泛素化并决定命运此外，无偏筛选产生了一种小分子 KPTN 激活剂，可增加 KPTN。蛋白质水平，抑制 mTORC1 活性，并增加溶酶体数量和活性。 KPTN 的药理学增强可减少体外 stau 蛋白聚集，具有治疗作用因此，KPTN 增强的净效应将有助于治疗阿尔茨海默病。通过激活自噬清除有毒蛋白质聚集体，这导致了中心假设： PDZRN3 对 KPTN 的控制影响神经元溶酶体活性，并且遗传或药理学 KPTN 的激活可能是减少 tau 蛋白聚集的一种途径，有两个目的将对此进行探讨。假设：（1）研究PDZRN3-KPTN轴对tau蛋白聚集的机制和生物学效应使用诱导多能干细胞 (iPSC) 和原代细胞模型进行体外实验，以及 (2) 检查这种机制并 Lear 博士还将利用遗传和药理学方法在 tau 传播动物模型中研究效果。结构化的培训计划，包括正式课程、会议和新技术的实践培训，在主要导师 Toren Finkel 博士、联合导师 Stacey Rizzo 博士和 Bill Chen 博士的指导下。