Cell Type Specific Genetic Manipulation to Dissect Cholinergic Interneuron Function and Plasticity in a Symptomatic Model of DYT1 Dystonia

细胞类型特异性基因操作剖析 DYT1 肌张力障碍症状模型中的胆碱能中间神经元功能和可塑性

• 批准号: 10548214
• 负责人: WILLIAM T. DAUER
• 金额: $ 50.49万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10548214
• 关键词: Ablation; Address; Adolescent; Autopsy; Autoreceptors; Behavioral; Biological Models; Brain; Brain Diseases; Cells; Chemicals; Corpus striatum structure; Data; Defect; Disease; Disease model; Drug usage; Dyskinetic syndrome; Dystonia; Electrophysiology (science); Elements; Functional disorder; Generations; Genetic; Genetic study; Glutamic Acid; Goals; Human; Interneuron function; Interneurons; L-DOPA induced dyskinesia; Literature; Machine Learning; Modeling; Morphology; Motor; Movement; Mus; Muscarinics; Nerve Degeneration; Neuronal Dysfunction; Neurons; Output; Parkinson Disease; Pathogenesis; Patients; Pharmaceutical Preparations; Publishing; Role; Signal Transduction; Slice; Symptoms; Synapses; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; TorsinA; cell type; cholinergic; cholinergic neuron; conditional knockout; design; designer receptors exclusively activated by designer drugs; effective therapy; genetic approach; genetic manipulation; loss of function mutation; motor behavior; motor disorder; multidisciplinary; nervous system disorder; neurodevelopment; neuron loss; neuropathology; novel; novel strategies; optogenetics; predictive modeling; prevent; targeted treatment; tomography; tool; translational study

Project Summary/Abstract Cholinergic neurons (ChIs) are a central but poorly understood element of striatal circuitry. A considerable literature strongly implicates ChI dysfunction in the pathogenesis of abnormal movements, especially in dystonia and levodopa-induced dyskinesias in Parkinson disease. A common theme of these studies is that maladaptive plastic changes cause aberrant ChI output and connectivity, promoting motor dysfunction. The central goal of this proposal is to advance understanding of the cellular and synaptic mechanisms through which ChIs cause motor dysfunction by employing novel selective genetic and chemical strategies in a recently validated model of DYT1 dystonia. Conditional Knock Out of torsinA from all striatal neurons (using Dlx5/6-Cre; “Dlx-CKO”) causes selective neurodegeneration of dorsolateral striatal ChI. ChI degeneration occurs roughly coincident with the juvenile onset of abnormal twisting movements in these mice, and selective ChI abnormalities are also present in postmortem tissue from DYT1 subjects. These movements are suppressed by the same anti-muscarinic compounds used to treat patients with DYT1 dystonia, establishing model therapeutic validity and suggesting shared pathophysiology with human dystonia. Surviving striatal ChIs are enlarged and hyperexcitable, and receive aberrant synaptic inputs. Selective ablation of these surviving ChI suppresses abnormal twisting, implicating these cells as key contributors to abnormal movements. Based on these data, we hypothesize that maladaptations in surviving ChIs drive motor dysfunction. Successful completion of the proposed studies will fundamentally advance understanding of maladaptive mechanisms whereby ChI function and connectivity drive abnormal movements, information highly significant for multiple striatal diseases. We will first address our hypothesis by testing the necessity of striatal ChI dysfunction in abnormal movement generation by selectively restoring torsinA to these cells (Aim 1), decisively moving beyond the current association between these factors. We will determine if cholinergic dysfunction arises primarily from intrinsic ChI abnormalities or defects in how they respond to afferents (Aim 2), and, informed by Aims 1 and 2, will pursue translational studies (Aim 3) testing whether directly modulating the activity of surviving ChIs can suppress dystonic-like movements. This proposal is therefore highly signifiant because it will define a circuit-based model of motor dysfunction that will inform the design of targeted therapeutics.

项目摘要/摘要 胆碱能神经元（CHIS）是纹状体回路的中心知识元素。一个很大的 文献强烈暗示了Chi功能障碍在异常运动的发病机理中，尤其是在肌张力障碍中 和左旋多巴引起的帕金森氏病的运动障碍。这些研究的一个共同主题是适应不良 塑料变化会导致异常的卡输出和连通性，从而促进电动机功能障碍。中心目标 该建议是提高对Chis引起的细胞和合成机制的理解 通过在最近经过验证的模型中采用新颖的选择性遗传和化学策略，通过运动功能障碍 Dyt1肌张力障碍。 从所有纹状体神经元（使用DLX5/6-CRE;“ DLX-CKO”）中敲出Torsina的有条件敲出会导致选择性 形式纹状体chi的神经变性。 Chi变性与少年发作大致相吻合 这些小鼠中异常的扭曲运动，以及验尸中也存在选择性卡的异常 来自Dyt1受试者的组织。这些运动被用于 治疗dyt1肌张力障碍患者，建立模型的热有效性并建议共享 人体肌张力障碍的病理生理学。幸存的纹状体chis可以增强且过度兴奋，并接受 异常突触输入。这些生存CHI的选择性消融抑制异常扭曲，牵涉 这些细胞是异常运动的关键因素。基于这些数据，我们假设 幸存的Chis Drive电动机功能障碍的疾病。 成功完成拟议的研究将从根本上提高对适应不良的理解 Chi功能和连接性驱动异常运动的机制，信息非常重要 用于多种纹状体疾病。我们将首先通过测试纹状体CHI的必要条件来解决我们的假设 通过选择性恢复这些细胞的躯干（AIM 1），果断地恢复躯干，在异常运动产生中功能障碍 超越这些因素之间的当前关联。我们将确定是否出现胆碱能功能障碍 固有的卡质异常或缺陷对传入的反应方式（AIM 2）的主要原理，并告知 目标1和2，将进行翻译研究（AIM 3）测试是否直接调节幸存的活动 Chis可以抑制肌张力障碍的运动。因此，该建议非常重要，因为它将定义 基于电路的运动功能障碍的模型将为目标治疗的设计提供信息。