TDP43 mRNA complex transport in physiologic and pathologic states

生理和病理状态下 TDP43 mRNA 复合物的转运

• 批准号: 9770568
• 负责人: Pallavi P. Gopal
• 金额: $ 18.52万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-01 至 2020-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9770568
• 关键词: 3' Untranslated Regions; Affect; Amyotrophic Lateral Sclerosis; Anatomy; Axon; Axonal Transport; Binding; Biological Assay; Biology; C-terminal; Cell Nucleus; Cellular Stress; Clinic; Clinical; Complex; Cytoplasm; Cytoplasmic Granules; Cytoplasmic Inclusion; DNA; DNA-Binding Proteins; Defect; Dendrites; Development Plans; Disease; Distal; Drosophila genus; Event; Exhibits; FMRP; Familial Amyotrophic Lateral Sclerosis; Fluorescent in Situ Hybridization; Fostering; Frontotemporal Dementia; Functional disorder; Genetic; Glycine; Grant; Image; Imaging Techniques; Impairment; Individual; International; Introns; Knowledge; Link; Mediating; Medical Genetics; Mentorship; Messenger RNA; Metabolism; Methods; Microtubules; Mitochondria; Modeling; Molecular; Motor; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neuroglia; Neurons; Neurosciences; Nuclear; Nuclear Export; Nuclear Localization Signal; Nuclear RNA; Organelles; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pennsylvania; Pharmacology; Physicians; Physiological; Play; Protein Dynamics; Proteins; RNA; RNA Binding; RNA Processing; RNA Recognition Motif; RNA Transport; RNA-Binding Proteins; RRM1 gene; RRM2 gene; Regulation; Research; Research Training; Resolution; Resources; Scientist; Signal Transduction; Stress; Structure; Synapses; Techniques; Technology; Temporal Lobe; Testing; Time; Toxic effect; Training; Training Programs; Transact; Transcript; Universities; Work; Yeasts; anterograde transport; base; career; career development; cell motility; collaborative environment; frontotemporal lobar dementia-amyotrophic lateral sclerosis; gain of function; innovation; knock-down; mRNA Precursor; mRNA Stability; meetings; messenger ribonucleoprotein; mutant; neuropathology; novel; parkin gene/protein; protein B; public health relevance; real-time images; response; single molecule; therapeutic target; trafficking

 DESCRIPTION (provided by applicant): This proposal describes a 5 year career development plan for Dr. Pallavi Gopal to serve as a transition to successful independent physician-scientist. Dr. Gopal completed her clinical training in Anatomic Pathology and Neuropathology at the University of Pennsylvania (Penn), and is now developing an independent research and training program that will allow her to gain expertise in spatial and temporal dynamics of mRNA transport under physiological conditions and in disease. This proposal brings together diverse resources in RNA metabolism, molecular neuroscience and neuropathology and will provide superb training for Dr. Gopal to develop into an independent physician-scientist. Research will be performed under the mentorship of Dr. Erika Holzbaur, an internationally recognized expert in microtubule-based motors and real-time axon transport dynamics. This grant will provide protected time for Dr. Gopal to gain expertise in neuronal cytoskeletal, organelle, and RNA-protein dynamics through formal coursework, scientific seminars and meetings. The collaborative environment at Penn will foster utilization of novel techniques to conduct the proposed project and will provide Dr. Gopal with the training required to proceed towards a successful academic career. Amyotrophic lateral sclerosis (ALS) and front temporal lobar degeneration (FTLD) exist on two ends of a clinic pathologic spectrum but share clinical, genetic, and pathologic features. Ubiquitinated cytoplasmic inclusions composed of Tran's active response DNA-binding protein of 43 kDa (TDP-43) are a shared feature of sporadic ALS and the most common form of FTLD; there is concomitant loss of normal nuclear TDP-43 expression. Moreover, the discovery of disease-linked mutations in TDP-43 and other RNA processing proteins highlights altered RNA metabolism as a common pathogenic mechanism of neurodegeneration. However, our knowledge of how TDP-43 mislocalization disrupts its nuclear and cytoplasmic RNA processing functions and/or mediates toxicity in the cytoplasm is still incomplete. The research plan will utilize innovative approaches with real-time imaging techniques in primary neurons to test two main hypotheses: that loss of nuclear TDP-43 results in reduced dynamic flux of TDP-43 target mRNA and proteins in axons and that cytoplasmic redistribution of TDP-43 under pathological conditions results in mislocalization of TDP-43- associated mRNA. The specific aims are to: 1) Determine whether loss of nuclear TDP-43 function reduces axonal trafficking of synaptic proteins and organelle turn over and 2) Determine how (A) loss of cytoplasmic TDP-43 RNA binding function and (B) stress-induced cytoplasmic TDP-43 aggregation affect localization and trafficking of mRNA in axons and dendrites. These studies will provide temporal and spatial resolution of individual RNA transcripts in neurons in order to gain a clearer understanding of altered RNA metabolism in ALS/FTLD pathogenesis.

 描述（由申请人提供）：该提案描述了 Pallavi Gopal 博士的 5 年职业发展计划，旨在过渡到成功的独立医师科学家。 Gopal 博士在宾夕法尼亚大学 (Penn) 完成了解剖病理学和神经病理学的临床培训，目前正在开发一项独立的研究和培训计划，该计划将使她能够获得生理条件和疾病下 mRNA 运输的空间和时间动力学方面的专业知识。该提案汇集了 RNA 代谢、分子神经科学和神经病理学方面的多种资源，将为 Gopal 博士发展成为一名独立的医师科学家提供极好的培训。研究将在国际公认的微管电机和实时轴突运输动力学专家 Erika Holzbaur 博士的指导下进行。这笔赠款将为 Gopal 博士提供受保护的时间，让他通过正式课程、科学研讨会和会议获得神经元细胞骨架、细胞器和 RNA 蛋白质动力学方面的专业知识。宾夕法尼亚大学的协作环境将促进新技术的利用来开展拟议的项目，并将为戈帕尔博士提供成功学术生涯所需的培训。 肌萎缩侧索硬化症 (ALS) 和前颞叶变性 (FTLD) 存在于临床病理谱的两端，但具有共同的临床、遗传和病理特征。由 43 kDa 的 Tran 活性反应 DNA 结合蛋白 (TDP-43) 组成的泛素化细胞质内含物是散发性 ALS 的共同特征，也是最常见的 FTLD 形式；正常核 TDP-43 表达同时丧失。此外，TDP-43 和其他 RNA 加工蛋白中与疾病相关的突变的发现凸显了 RNA 代谢的改变是神经退行性疾病的常见致病机制。然而，我们对 TDP-43 错误定位如何破坏其核和细胞质 RNA 加工功能和/或介导细胞质毒性的了解仍然不完整。该研究计划将利用原代神经元实时成像技术的创新方法来测试两个主要假设：核TDP-43的丢失导致轴突中TDP-43靶mRNA和蛋白质的动态通量减少，以及病理条件下TDP-43的细胞质重新分布导致TDP-43相关mRNA的错误定位。具体目标是：1) 确定核 TDP-43 功能的丧失是否会减少突触蛋白的轴突运输和细胞器更新；2) 确定 (A) 细胞质 TDP-43 RNA 结合功能的丧失和 (B) 应激诱导的细胞质 TDP-43 聚集如何影响轴突和树突中 mRNA 的定位和运输。这些研究将提供神经元中单个 RNA 转录本的时间和空间分辨率，以便更清楚地了解 ALS/FTLD 发病机制中 RNA 代谢的改变。