Study of AR transcriptional network in stem cell model of SBMA

SBMA干细胞模型中AR转录网络的研究

基本信息

批准号：
10373083
负责人：
Helen C Miranda
金额：
$ 39.44万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10373083
关键词：
AR gene Adult Affect Androgen Receptor Androgens Animal Model Animals Atrophic Biological Assay CRISPR interference Cells Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Consensus DNA Data Data Set Devices Disease Electrophysiology (science)Engineering Etiology Event Family Fertility Functional disorder Gene Expression Genes Genetic Genetic Transcription Genomics Glutamine Goals Gynecomastia Hi-C Human Huntington Disease In Vitro Inherited Kennedy Syndrome Ligands Link Literature Luciferases Microfluidics Mitochondria Molecular Motor Activity Motor Neuron Disease Motor Neurons Muscle Muscle Cells Muscle Contraction Muscle Weakness Mutation Neuraxis Neurodegenerative Disorders Neuromuscular Diseases Neuromuscular Junction Neuronal Dysfunction Pathogenesis Pathogenicity Pathology Pathway interactions Patients Phenotype Quantitative Reverse Transcriptase PCR Research Resources Rhodopsin Skeletal Muscle Stanolone Synapsins System Therapeutic Transcriptional Regulation Validation Work alpha Bungarotoxin cell type chromatin immunoprecipitation deep sequencing defined contribution differential expression genome-wide human stem cells induced pluripotent stem cell member motor control motor neuron degeneration multi-electrode arrays mutant nervous system disorder neurofilament neuromuscular function optogenetics overexpression polyglutamine promoter receptor binding spinal and bulbar muscular atrophy stem cell model theories therapy development tool transcription factor transcriptome sequencing transcriptomics

项目摘要

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.

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，其特征是成人发病的近端肌肉无力，原因是 SBMA 患者表现出雄激素不敏感的迹象，包括男性乳房发育症、生育能力下降和睾丸萎缩，都是由 CAG- 引起的。多聚谷氨酰胺 (polyQ) 在雄激素受体 (AR) 基因中重复扩增，是其中之一包括亨廷顿舞蹈病在内的九种 CAG-polyQ 重复疾病家族的成员。几十年来，对神经系统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，在过去的十年中，已经出现了这种情况。运动神经元疾病领域越来越多的证据挑战了流行的神经中心论许多神经系统疾病的病因学理论在 SBMA 中越来越多。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。治疗目的，然而，不同的群体已经证明了成功的治疗方法使用不同的 SBMA 动物针对骨骼肌或中枢神经系统通过对模型进行检查，文献中缺乏共识强调了研究的必要性。 SBMA AR 突变对受影响细胞类型（骨骼肌和运动神经元）的影响我们发现，polyQ 突变会导致 AR 与 DNA 导致转录失调，可能在运动神经元中致病，因此，该提案的目标是将 AR 基因组结合起来。由 SBMA 和 CRISPR 工程生成的占用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，然后我们将共培养。 iPSC 衍生的骨骼肌与 iPSC 衍生的运动神经元来调节 AR 目标在 AR 转录网络中识别并通过以下方式研究神经肌肉接头 (NMJ) 分析运动神经元和骨骼肌的电生理活动。将增进对人类突变体 AR 至 SBMA 分子机制的理解发病机制并评估 iPSC 衍生的骨骼肌和运动神经元工具的效用开展 SBMA 体外研究。