Dissecting the in vivo role of Huntingtin in Rab vesicle movement on microtubules

剖析亨廷顿蛋白在微管上 Rab 囊泡运动中的体内作用

基本信息

批准号：
8721495
负责人：
Shermali Gunawardena
金额：
$ 7.73万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-15 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8721495
关键词：
Actins Affect Axon Axonal Transport Binding Brain Diseases Caliber Cell Death Chorea Clathrin Complex Corpus striatum structure Cytoplasmic Inclusion Cytoskeleton Defect Development Disease Disease Pathway Drosophila genus DsRed Dynein ATPase Endocytosis Endosomes Event FDA approved Genetic Gliosis Goals Health Human Huntington Disease Impaired cognition Inherited Intracellular Transport Investigation Kinesin Knowledge Life Mediating Membrane Protein Traffic Microtubule Proteins Microtubules Mission Motor Movement Mus Nerve Nerve Degeneration Neurodegenerative Disorders Neuronal Dysfunction Neurons Nuclear Inclusion Organism Orthologous Gene Outcome Pathology Pathway interactions Physiological Play Preventive Intervention Proteins Public Health Recycling Regulation Research Role Symptoms Testing Therapeutic Therapeutic Intervention Transport Vesicles Vesicle Work base burden of illness cell motility disability effective therapy end stage disease genetic analysis human Huntingtin protein in vivo innovation loss of function mutant neuron loss neuropathology novel prevent public health relevance

项目摘要

DESCRIPTION (provided by applicant): Huntington's disease (HD) is a devastating, dominantly inherited neurodegenerative disease clinically characterized by chorea and cognitive impairment due to loss of striatal neurons. Currently there are no effective treatments/cures for HD. Most therapeutic treatments currently used are aimed at dissolving/dissociating aggregates and preventing cell death, common neuropathology seen at the end stage of disease. Although the HD protein, huntingtin (HTT) is critical for viability, the complexity of HTT-mediated associations indicates multiple functions. Thus the challenge is to unravel the primary function of HTT, which when disrupted initiates disease. Previous work put forth a tantalizing proposal that disruption of axonal transport within long, narrow-caliber axons is an early event that causes protein accumulations that elicit cell death, ultimately resulting in neuronal dysfunction observed in HD. Our long-term goal is to understand how HTT-mediated axonal transport defects initiate disease pathways. The objective here, which is our next step in the pursuit of thi goal, is to determine how HTT influences the transport of a specific sub class of vesicles (Rab proteins). Our central hypothesis is that disruption of Rab vesicle transport within axons mediated by HTT can contribute to early neuropathology observed in HD. There are two clear predictions of this hypothesis; 1: HTT and Rab proteins are on the same vesicles and 2: Rab vesicles use kinesin-1 and dynein motors for movement on microtubules (MT). In this context our specific aim is to identify how HTT influences Rab proteins for MT-dependent transport within axons. We have 5 specific objectives, 1: determine how HTT influences Rab proteins, 2: test the prediction that HTT and Rab11, Rab32 and RabX4 are on the same vesicle, 3: test the prediction that Rab32 and RabX4 are both on the Rab11 vesicle, 4: test the prediction that Rab32 and RabX4 use kinesin-1 and dynein motors for movement on MT, and 5: test the prediction that mutant HTT disrupts Rab-mediated functions. A comprehensive in vivo approach will be used to dissect the physiological role of HTT in Rab vesicle transport in an organism. The rationale for the proposed research is that once the mechanisms of how HD disease is initiated by perturbations in Rab transport by mutant HTT are known, new and innovative approaches against HD can be developed. Therefore identifying how HTT normally functions in neurons will have a significant impact on providing novel target pathways for developing effective preventive and therapeutic interventions, which are currently unavailable for HD. Thus our work is innovative, in our opinion because it represents a new and substantive departure from the status quo, namely the approach of detailing the role of HTT using in vivo dynamics of vesicle movement in a living organism. The proposed research is significant, because it is expected to vertically advance and expand our understanding of how disease pathways initiate, which will significantly alter current knowledge. The knowledge acquired will dramatically propel the development of numerous pharmacological or genetic modifiers against axonal defects or to restore Rab function.

描述（由申请人提供）：亨廷顿氏病（HD）是一种毁灭性的，主要遗传的神经退行性疾病，其临床特征是由于纹状体神经元的丧失而导致的唱片和认知障碍。目前尚无高清治疗方法/治疗方法。当前使用的大多数治疗性治疗旨在溶解/解离骨料并预防细胞死亡，这是在疾病末期可见的常见神经病理学。尽管HD蛋白，Huntingtin（HTT）对于生存力至关重要，但HTT介导的关联的复杂性表明多种功能。因此，挑战是揭示HTT的主要功能，当破坏疾病时。先前的工作提出了一项诱人的建议，即在长狭窄的轴突内破坏轴突转运是一个早期事件，导致蛋白质积累引起细胞死亡，最终导致HD中观察到的神经元功能障碍。我们的长期目标是了解HTT介导的轴突运输缺陷如何启动疾病途径。这里的目标是我们追求目标的下一步，是确定HTT如何影响特定子类囊泡（Rab蛋白）的运输。我们的中心假设是，HTT介导的轴突内的Rab囊泡转运破坏可以有助于在HD中观察到的早期神经病理学。关于这一假设有两个明确的预测。 1：HTT和RAB蛋白在相同的囊泡上，2：Rab囊泡使用驱动蛋白1和动力蛋白电动机在微管上运动（MT）。在这种情况下，我们的具体目的是确定HTT如何影响轴突内MT依赖性转运的RAB蛋白。 We have 5 specific objectives, 1: determine how HTT influences Rab proteins, 2: test the prediction that HTT and Rab11, Rab32 and RabX4 are on the same vesicle, 3: test the prediction that Rab32 and RabX4 are both on the Rab11 vesicle, 4: test the prediction that Rab32 and RabX4 use kinesin-1 and dynein motors for movement on MT, and 5: test the prediction that mutant HTT破坏RAB介导的功能。一种全面的体内方法将用于剖析HTT在生物体中HTT的生理作用。拟议的研究的理由是，一旦已知通过突变体HTT在Rab运输中扰动引发HD疾病的机制，就可以开发针对HD的新和创新方法。因此，确定HTT通常在神经元中的功能将对提供有效的预防性和治疗性干预措施的新目标途径产生重大影响，而HD目前无法使用HD。因此，我们认为我们的工作具有创新性，因为它代表了与现状的新事物，即详细介绍使用囊泡运动在生物体中使用体内动力学的作用的方法。拟议的研究很重要，因为它有望垂直提高和扩展我们对疾病途径如何发起的理解，这将显着改变当前的知识。获得的知识将极大地推动许多药理学或遗传修饰剂的开发免受轴突缺陷或恢复RAB功能。