Histone deacetylases - therapeutic targets for functional restoration after strok

组蛋白脱乙酰酶 - 中风后功能恢复的治疗靶点

基本信息

批准号：
8048964
负责人：
RICHARD S MORRISON
金额：
$ 33.44万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8048964
关键词：
Accounting Acute Adult Affect Anatomy Animal Model Animals Antigens Apoptosis Apoptotic Aspirin Autopsy Biological Preservation Brain Brain region Bromodeoxyuridine Cause of Death Cell Culture Techniques Cell Death Cell Survival Cells Cerebral Ischemia Cessation of life Clinical Treatment Clinical Trials Corpus striatum structure Cyan Fluorescent Protein Data Development Enzymes FDA approved Fiber Frequencies Funding Opportunities Generations Glucose Health Resources Histologic Histone Acetylation Histone Deacetylase Histone Deacetylase Inhibitor Histone deacetylase inhibition Histones Human In Situ In Vitro Individual Infarction Injury Intervention Ischemia Ischemic Stroke Lead MS-275 Malignant Neoplasms Mediating Methods Middle Cerebral Artery Occlusion Mitochondria Mitotic Modeling Monitor Morphology Motor Mus Myelin Natural regeneration Nervous System Physiology Neuronal Differentiation Neurons Optic Nerve Outcome Outcome Measure Oxygen Pathology Pharmaceutical Preparations Preparation Process Production Proteins Recovery Recovery of Function Research Safety Sensory Stem cells Stroke Subfamily lentivirinae Testing Therapeutic Tissues Transgenic Mice Transgenic Organisms Transient Cerebral Ischemia Vorinostat brain repair cellular transduction cognitive recovery density deprivation design disability drugged driving enzyme activity excitotoxicity functional outcomes functional restoration gray matter improved inhibitor/antagonist interest killings knock-down meetings nerve stem cell neurogenesis neuron loss neuronal cell body neuroprotection newborn neuron novel strategies post stroke postnatal progenitor protective effect public health relevance research study response restoration selective expression small hairpin RNA therapeutic target thrombolysis white matter

项目摘要

DESCRIPTION (provided by applicant): Finding interventions that improve outcome from ischemic stroke has proved to be challenging and current treatment is limited to thrombolysis, aspirin, and management in a stroke unit. An ideal stroke therapeutic would minimize damage to mature neurons and white matter, and also maximize the generation of new neurons from endogenous progenitors. Recent findings of our own and of others, both in vitro and in animal models of stroke, suggest that histone deacetylase (HDAC) inhibitors meet these criteria. Drug administration protects isolated neurons from induced apoptotic cell death and white matter from oxygen-glucose deprivation, results in improved histologic and functional outcomes following cerebral ischemia, and appears to drive 'neuronal' differentiation of multipotent neural progenitor cells. Therefore, we hypothesize that HDAC inhibition in stroke may have acute effects (to ameliorate white matter excitotoxicity), and in the intermediate and longer term, added benefit by reducing apoptosis and encouraging regeneration. As a number of HDAC inhibitors are either already approved by the FDA (vorinostat), or well-advanced in clinical trials (MS-275) for other reasons, then re- purposing one or more of these drugs for stroke could be expedited. In addition, development of new and specific inhibitors is ongoing, especially against the Class I HDACs which are the focus of interest here. Studies described in Aim 1 are designed to identify the specific HDACs which account for the beneficial actions of these inhibitors, using in vitro and ex vivo preparations in which individual HDACs are knocked down with shRNA and transduced cells/tissues then subjected to oxygen-glucose deprivation. Having identified specific HDACs, we shall explore in Aim 2 the potential substrates and mechanisms responsible for the beneficial actions of HDAC inhibition. Finally, in the last aim we consider whether treatment of mice with MS-275, a Class I HDAC inhibitor, preserves anatomic integrity and promotes long term functional (motor, sensory, cognitive) recovery from transient cerebral ischemia (middle cerebral artery occlusion), and whether restoration of function correlates with the extent of 'neuronogenesis'. Two recently developed transgenic lines, p53+/+ and p53 -/- mitoCFP mice, will be employed so that we can monitor ischemic pathology in CFP+ fiber tracts, assess drug-associated changes in mitochondrial frequency and distribution within neuronal cell bodies and their processes, and also determine whether HDAC inhibition ultimately involves targets in addition to neuronal p53. PUBLIC HEALTH RELEVANCE: This proposal is submitted in response to PA-08-099 Mechanisms of functional recovery after stroke, "a funding opportunity to promote research to understand the processes of brain repair that lead to functional recovery in order to develop methods to optimize existing practices and to develop new approaches to improve post-stroke outcomes". Currently, there is no drug available to enhance neuroprotection and restore function following human stroke. Experimental animal studies suggest that broad inhibition of histone deacetylase (HDAC) activities in the brain following injury results in neuronal protection and also promotes the generation of new neurons. The purpose of the studies outlined in this proposal is to identify which HDACs are involved in order to select specific drugs to target these particular enzymes. Selected drugs will be tested to reveal what functional benefits they confer on the recovery of mice from stroke injury.

描述（由申请人提供）：事实证明，寻找改善缺血性中风结果的干预措施具有挑战性，目前的治疗仅限于溶栓、阿司匹林和中风病房的管理。理想的中风治疗方法应最大限度地减少对成熟神经元和白质的损害，并最大限度地从内源性祖细胞中生成新神经元。我们自己和其他人最近在体外和中风动物模型中的发现表明，组蛋白脱乙酰酶 (HDAC) 抑制剂符合这些标准。药物给药可以保护分离的神经元免受诱导的细胞凋亡和白质免受氧-葡萄糖剥夺的影响，从而改善脑缺血后的组织学和功能结果，并且似乎可以驱动多能神经祖细胞的“神经元”分化。因此，我们假设 HDAC 抑制在中风中可能具有急性作用（改善白质兴奋性毒性），并且在中长期来看，通过减少细胞凋亡和促进再生而增加益处。由于许多 HDAC 抑制剂要么已获得 FDA 批准（伏立诺他），要么因其他原因在临床试验（MS-275）中取得进展，因此可以加快将这些药物中的一种或多种重新用于中风。此外，新的特异性抑制剂的开发正在进行中，特别是针对 I 类 HDAC 的抑制剂，这是本文关注的焦点。目标 1 中描述的研究旨在使用体外和离体制剂来鉴定导致这些抑制剂有益作用的特定 HDAC，其中用 shRNA 敲低单个 HDAC 并转导的细胞/组织，然后进行氧-葡萄糖剥夺。确定了特定的 HDAC 后，我们将在目标 2 中探索导致 HDAC 抑制的有益作用的潜在底物和机制。最后，在最后一个目标中，我们考虑用 MS-275（一种 I 类 HDAC 抑制剂）治疗小鼠是否可以保留解剖完整性并促进短暂性脑缺血（大脑中动脉闭塞）的长期功能（运动、感觉、认知）恢复，以及功能的恢复是否与“神经元发生”的程度相关。两个最近开发的转基因系，p53+/+ 和 p53 -/- mitoCFP 小鼠，将被用来监测 CFP+ 纤维束的缺血病理学，评估神经元细胞体内线粒体频率和分布及其过程的药物相关变化，并确定 HDAC 抑制最终是否涉及神经元 p53 之外的靶点。公共健康相关性：本提案是为了响应 PA-08-099 中风后功能恢复机制而提交的，“这是一个资助机会，旨在促进研究以了解导致功能恢复的大脑修复过程，以便开发优化现有方法的方法”实践并开发新方法来改善中风后的结果”。目前，尚无药物可用于增强人类中风后的神经保护和恢复功能。实验动物研究表明，损伤后大脑中组蛋白脱乙酰酶 (HDAC) 活性的广泛抑制会导致神经元保护，并促进新神经元的生成。本提案中概述的研究目的是确定涉及哪些 HDAC，以便选择针对这些特定酶的特定药物。选定的药物将进行测试，以揭示它们对中风损伤小鼠的恢复有何功能益处。