Modulation of striatal cholinergic interneuron activity to prevent dystonic cerebral palsy

项目总结 这一建议将确定在发育过程中纹状体胆碱能中间神经元(CHI)活性是否增加 小鼠脑可预防新生儿脑损伤后肌张力障碍 。新生儿肌张力障碍型脑瘫(CP) 脑损伤是导致儿童肌张力障碍的最常见原因，通常是难治性和功能性的。 让人虚弱。然而，其独特的病理生理机制仍未得到充分研究。肌张力障碍的病理生理学更常见 在以纹状体CHI过度兴奋为特征的罕见遗传性肌张力障碍模型中进行研究。然而， 抗胆碱能药物治疗慢性阻塞性肺病的肌张力障碍常常无效。确定是否存在纹状体 针对肌张力障碍CP的胆碱能病理可以产生更好的靶向治疗。为此，我开发了 一种临床相关的新生儿缺氧性脑损伤的啮齿动物模型 损伤后三周肌张力障碍，模拟新生儿脑损伤和肌张力障碍之间的临床潜伏期 浮现。这一潜伏期允许对预防肌张力障碍的症状前干预措施进行测试。我的 初步数据显示，在我的模型中，纹状体CHI数量增加，但年轻人的纹状体CHI兴奋 小鼠在症状前窗口可能对肌张力障碍有保护作用。总而言之，这些数据表明 纹状体CHI数量增加和纹状体CHI过度兴奋可能是一种代偿机制 因此，可以加强对肌张力障碍的保护，以预防新生儿脑损伤后的肌张力障碍。 为了验证这一假设，我提出了以下目标：(1)确定纹状体的化学发生调制是否 新生小鼠脑损伤后幼鼠CHI活性改变成年小鼠肌张力障碍严重程度；(2)测定 幼年、健康小鼠纹状体CHI活性的化学发生调控是否会引起肌张力障碍 以及(3)确定在遗传性肌张力障碍中观察到的纹状体CHI高兴奋性是否也 出现在我的新生儿脑损伤后肌张力障碍模型中。这些研究将确定是否会预先 新生儿脑损伤后有症状地增加纹状体CHI放电可减轻或预防肌张力障碍。 我的长期职业目标是经营一家专注于预防性治疗开发的转化型研究实验室 治疗肌张力障碍的CP。我研究了十年的基底节病理生理学，并开发了一种新的模型 新生儿脑损伤后的肌张力障碍，将用于拟议的实验。然而，要完成 建议的研究并促进我向独立的过渡，我需要在Slice方面进行额外的指导培训 电生理学(Steve Mennerick博士)和化学遗传学(Jordan McCall博士)。作为我的内科科学家 顾问乔尔·珀尔穆特博士将提供肌张力障碍病理生理学方面的专业知识，并确保翻译 我的研究的相关性。华盛顿大学医学院和神经病学系提供 世界知名的研究环境和热情而有效地支持初级教员的遗产。 总之，我提出的研究、指导团队、培训计划和机构环境为我铺平了道路 R01的独立性和呈交 确定治疗目标 治疗肌张力障碍的CP。

PROJECT SUMMARY This proposal will determine whether increasing striatal cholinergic interneuron (ChI) activity in the developing mouse brain can prevent dystonia following neonatal brain injury . Dystonic cerebral palsy (CP) due to neonatal brain injury is the most common cause of childhood dystonia and is often medically refractory and functionally debilitating. Yet, its unique pathophysiology remains understudied. Dystonia pathophysiology is more commonly studied in models of rare genetic dystonias which are characterized by striatal ChI hyperexcitability. However, anticholinergic medications are often ineffective for treating dystonia in CP. Determining whether there is striatal cholinergic pathology specific to dystonic CP could yield better targeted treatments. To this end, I have developed a clinically-relevant rodent model of neonatal hypoxic brain injury that displays electrophysiologic markers of dystonia three weeks after injury, mimicking the clinical latency period between neonatal brain injury and dystonia emergence. This latency period allows testing of pre-symptomatic interventions for dystonia prevention. My preliminary data demonstrate increased striatal ChI number in my model but that striatal ChI excitation in young mice during the pre-symptomatic window may be protective against dystonia. In sum, these data suggest that increased striatal ChI number and striatal ChI hyperexcitability may be compensatory mechanisms that are protective against dystonia and, therefore, could be enhanced to prevent dystonia following neonatal brain injury. To test this hypothesis, I propose the following aims: (1) determine whether chemogenetic modulation of striatal ChI activity in young mice after neonatal brain injury changes dystonia severity in adult mice; (2) determine whether chemogenetic modulation of striatal ChI activity in young, otherwise healthy, mice can cause dystonia in adult mice; and (3) determine whether the striatal ChI hyperexcitability observed in genetic dystonias is also present in my model of dystonia following neonatal brain injury. These studies will determine whether pre- symptomatically increasing striatal ChI firing after neonatal brain injury could reduce or prevent dystonia. My long-term career goal is to run a translational research lab focused on preventative treatment development for dystonic CP. I have studied basal ganglia pathophysiology for ten years and have developed a new model of dystonia following neonatal brain injury which will be used for the proposed experiments. However, to complete the proposed research and facilitate my transition to independence, I need additional mentored training in slice electrophysiology (Dr. Steve Mennerick) and chemogenetics (Dr. Jordan McCall). As my physician-scientist advisor, Dr. Joel Perlmutter will provide expertise in dystonia pathophysiology and ensure the translational relevance of my research. The Washington University School of Medicine and Department of Neurology provide a world-renowned research environment and a legacy of passionately and effectively supporting junior faculty. In sum, my proposed research, mentorship team, training plan, and institutional environment pave my path to independence and submission of an R01 on identification of treatment targets for dystonic CP.