Study of AR transcriptional network in stem cell model of SBMA

X连锁脊髓和延髓肌萎缩症(也称为SBMA或肯尼迪病)是一种 罕见的神经肌肉疾病，其特征是成人起病的近端肌肉无力，原因是 低位运动神经元变性。SBMA患者表现出雄激素不敏感的迹象， 包括女性乳房发育、生育力下降和睾丸萎缩。SBMA是由CAG引起的- 多聚谷氨酰胺(PolyQ)重复扩增在雄激素受体(AR)基因中，是成员之一 包括亨廷顿病在内的一个九口之家的CAG-PolyQ重复疾病。为 几十年来，对神经疾病基础的研究集中在 神经元功能障碍对疾病发病机制的影响。然而，在过去的十年里， 运动神经元疾病领域越来越多的证据挑战了盛行的神经中心论 许多神经疾病的病因学理论。。在SBMA的情况下， 有证据表明肌肉功能障碍是疾病发病机制的主要组成部分。为 治疗目的，然而，不同的小组已经证明了成功的治疗方法 使用不同的SBMA动物靶向骨骼肌或中枢神经系统 模特们。因此，文献中缺乏共识凸显了研究的必要性 受影响的细胞类型-骨骼肌和运动神经元-的SBMA AR突变 人类背景。我们假设PolyQ突变会导致AR与 DNA导致转录失调，这可能是运动神经元的致病因素， 骨骼肌或两者兼而有之。因此，这项建议的目标是在全基因组范围内结合AR SBMA和CRISPR工程生成的占有率和基因表达数据集 同基因控制IPSC衍生的骨骼肌和运动神经元。然后，我们将共同培养 IPSC来源的骨骼肌与IPSC来源的运动神经元一起调节AR靶点 在AR转录网络中识别并研究神经肌肉连接(NMJ) 分析运动神经元和骨骼肌的电生理活动。这部作品 将促进对人突变型AR对SBMA分子机制的理解 IPSC来源的骨骼肌和运动神经元工具的致病机制和实用性评价 开展SBMA的体外研究。

X-linked spinal and bulbar muscular atrophy (also known as SBMA or Kennedy's disease) is a rare neuromuscular disorder characterized by adult-onset proximal muscle weakness due to lower motor neuron degeneration. SBMA patients display signs of androgen insensitivity, including gynecomastia, reduced fertility, and testicular atrophy. SBMA, is caused by a CAG- polyglutamine (polyQ) repeat expansion in the androgen receptor (AR) gene and is one member of a family of nine CAG-polyQ repeat disorders that includes Huntington’s disease. For decades, research into the basis of neurological disease focused upon the contribution of neuronal dysfunction to disease pathogenesis. However, over the last ten years, there has been growing evidence in the motor neuron disease field that challenges the prevailing neurocentric theory of the etiology of many neurological diseases. . In the case of SBMA, there is increasing evidence implicating muscle dysfunction as a major component of disease pathogenesis. For therapeutic purposes, however, different groups have demonstrated successful treatments targeting either the skeletal muscles or the central nervous system using different SBMA animal models. Consequently, this lack of consensus in the literature underscores the need for studies of the SBMA AR-mutation on affected cell types - skeletal muscles and motor neurons - in a human background. We hypothesized that polyQ mutation cause disruption in AR binding to the DNA which leads to transcriptional dysregulation that can be pathogenic in motor neurons, skeletal muscles or both. Therefore, the goal of this proposal is to combine AR genome wide occupancy and gene expression data sets generated from SBMA and CRISPR engineered isogenic controls iPSC-derived skeletal muscle and motor neurons. We will then, co-culture the iPSC-derived skeletal muscles with iPSC-derived motor neurons to modulate the AR targets identified in the AR transcriptional network and investigate neuromuscular junctions (NMJ) by analyzing electrophysiological activity of the motor neurons and the skeletal muscles. This work will advance understanding on the molecular mechanisms of human mutant AR to SBMA pathogenesis and evaluate the utility of iPSC-derived skeletal muscles and motor neurons tool to develop SBMA in vitro studies.