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

基本信息

批准号：
9013735
负责人：
Pallavi P. Gopal
金额：
$ 18.94万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9013735
关键词：
3&apos 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 Dementia Dendrites Development Plans Disease Distal Drosophila genus Event Exhibits FMRP Familial Amyotrophic Lateral Sclerosis Fluorescent in Situ Hybridization Fostering Frontotemporal Lobar Degenerations Functional disorder Genetic Glycine Grant Image Imaging Techniques Individual Introns Knowledge Life Link Lobar 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 Pathologic Pathology Pennsylvania 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 Synapses Techniques Technology Testing Time Toxic effect Training Training Programs Transcript Universities Work Yeasts anterograde transport base career career development cell motility collaborative environment gain of function innovation mRNA Precursor mRNA Stability meetings messenger ribonucleoprotein mutant neuropathology novel parkin gene/protein parkin protein protein B protein TDP-43 public health relevance response single molecule therapeutic target trafficking

项目摘要

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（由应用程序提供）：该提案描述了Pallavi Gopal博士为5年的职业发展计划，以作为成功独立的身体科学家的过渡。 Gopal博士在宾夕法尼亚大学（PENN）完成了她在解剖病理学和神经病理学方面的临床培训，现在正在制定一个独立的研究和培训计划，使她能够在生理状况和疾病中获得mRNA运输的空间和临时动力学专业知识。该提案汇集了RNA代谢，分子神经科学和神经病理学中的潜水资源，并将为Gopal博士提供出色的培训，以发展成独立的身体科学家。研究将在基于微管电机和实时轴突运输动力学方面的国际认可的专家Erika Holzbaur博士的心态下进行。这项赠款将为Gopal博士提供受保护的时间，以通过正式的课程，科学的半身和会议获得神经元细胞骨架，细胞器和RNA-蛋白质动力学的专业知识。宾夕法尼亚州的协作环境将促进新技术来开展拟议项目，并将为Gopal博士提供成功的学术职业所需的培训。肌萎缩性侧索硬化症（ALS）和前临时LOBAR变性（FTLD）存在于临床病理谱的两端，但具有临床，遗传和病理特征。由TRAN的活性反应DNA结合蛋白（TDP-43）组成的泛素化细胞质夹杂物是零星ALS的共同特征，是FTLD的最常见形式的共享特征。正常核TDP-43表达伴随丧失。此外，在TDP-43和其他RNA加工蛋白中发现疾病连接的突变突出了RNA代谢改变是神经变性的常见致病机制。但是，我们对TDP-43错误定位如何破坏其核和细胞质RNA处理功能和/或介导细胞质中的毒性的了解仍然不完整。该研究计划将利用主要神经元中实时成像技术的创新方法来检验两个主要假设：核TDP-43的丧失导致TDP-43靶标mRNA和蛋白质中的动态通量降低，并且在病理学下，TDP-43的细胞质量重新分布在MISLOCTOLICAD-4的情况下，TDP-43的细胞质量重新分布。具体目的是：1）确定核TDP-43功能的丧失是否会降低突触蛋白和机构的轴突运输，而2）确定（a）细胞质TDP-43 RNA结合功能的损失以及（B）压力诱导的TDP-43聚集量会影响MRMNA和thratike in MRNNA in MRNNA in MRNNA in MRNNA in MRNNA in MRNNA in MRNNA in MRNNA in MRNNA and NRANNA。这些研究将提供神经元中单个RNA转录本的暂时和空间分辨率，以便对ALS/FTLD发病机理中RNA代谢的改变更清晰地理解。