Study of SBMA mutant AR transcriptional network in stem cell-derived motor neurons and skeletal muscle

干细胞源性运动神经元和骨骼肌中SBMA突变体AR转录网络的研究

• 批准号: 10599883
• 负责人: Helen C Miranda
• 金额: $ 17.48万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10599883
• 关键词: AR gene; Address; Adult; Affect; Androgen Receptor; Androgens; Animal Model; Atrophic; Award; Biological Assay; CRISPR interference; Cell Death; Cells; Central Nervous System; ChIP-seq; Consensus; Data; Data Set; Disease; Disease model; Etiology; Family; Fertility; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Glutamine; Goals; Grant; Gynecomastia; Hi-C; Human; Huntington Disease; In Vitro; Inherited; International; Journals; Kennedy Syndrome; Letters; Ligands; Link; Literature; Luciferases; Manuscripts; Mentors; Mentorship; Mitochondria; Molecular; Motor Neuron Disease; Motor Neurons; Muscle; Muscle Weakness; Mutation; Neurodegenerative Disorders; Neuromuscular Diseases; Neuronal Dysfunction; Neurosciences; Pathogenesis; Pathology; Pathway interactions; Patients; Phenotype; Quantitative Reverse Transcriptase PCR; Reporter Genes; Research; Scientist; Skeletal Muscle; Specific qualifier value; Stanolone; Testing; Therapeutic; Time; Tissues; Toxic effect; Training; Transcriptional Regulation; Universities; Validation; Work; cell type; chromatin immunoprecipitation; comparison control; deep sequencing; defined contribution; differential expression; experience; functional genomics; gene repression; genome-wide; human stem cells; induced pluripotent stem cell; member; motor control; motor neuron degeneration; mutant; nervous system disorder; overexpression; polyglutamine; professor; programs; responsible research conduct; skeletal muscle wasting; spinal and bulbar muscular atrophy; stem cell biology; stem cell model; stem cells; symposium; theories; therapy development; tool; transcription factor; transcriptome sequencing; transcriptomics

SUMMARY 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. Recently, we have identified increased cell death in control motor neurons subjected to conditioned media from SBMA iPSC-derived skeletal muscles compared to control derived skeletal muscles. This finding emphasizes the importance of muscle toxicity in SBMA disease pathogenesis. For therapeutic purposes, however, there is lack of consensus in the literature. Different groups have been able to demonstrate successful treatments targeting either the skeletal muscle or the central nervous system using different SBMA animal models. Consequently, there is a need for studies of the SBMA AR-mutation on the affected cell types, skeletal muscle and motor neurons, in a human background. Therefore, the applicant, Dr. Helen C. Miranda, is proposing to combine her considerable experience in stem cell biology and motor neuron disease modeling to a mentorship in functional genomics, to test the hypothesis that the AR transcriptional network is tissue-specific in SBMA. This project will test this hypothesis by combining AR genome wide occupancy and gene expression data sets generated from SBMA and isogenic controls iPSC-derived skeletal muscle and motor neurons. 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. Dr. Miranda is a new Assistant Professor in the Department of Genetics and the Department of Neuroscience at Case Western Reserve University. She will devote 75% of her time to research under this award and will supplement her research with didactic training in genomic and transcriptomic analyses. This training will be comprised of 1) departmental and university courses, 2) seminars and journal clubs 3) responsible conduct of research courses and 4) national and international conferences. Dr. Miranda will be mentored by Dr. Anthony Wynshaw-Boris and Dr. Ann Harris at Case Western Reserve University. These established scientists are both renowned experts in stem cell biology and functional genomics. Dr. Miranda has met with each of her mentors to discuss this project and will continue to meet with them at regular intervals (specified in mentorship letters) during the course of this award. She is expected to produce manuscripts as corresponding or co-corresponding author and be competitive for R-level grants during the course of this award. This project will integrate Dr. Miranda’s current expertise with additional training to develop a well-rounded, independent research program.

摘要 X连锁脊髓和延髓肌萎缩症(也称为SBMA或肯尼迪病)是一种罕见的疾病 以成人起病的近端肌肉无力为特征的神经肌肉疾病，由较低的运动神经元引起 退化。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是由雄激素中CAG-聚谷氨酰胺(PolyQ)重复扩增引起的 受体(AR)基因，是九种CAG-PolyQ重复疾病家族的成员之一，该家族包括 亨廷顿氏病。几十年来，对神经系统疾病基础的研究主要集中在 神经元功能障碍在疾病发病机制中的作用。然而，在过去的十年里， 运动神经元疾病领域中越来越多的证据挑战了盛行的神经中心理论 许多神经疾病的病因学。最近，我们发现在控制运动中细胞死亡增加。 应用SBMA IPSC来源骨骼肌条件培养液的神经元与对照组的比较 衍生的骨骼肌。这一发现强调了肌肉毒性在SBMA病中的重要性。 发病机制。然而，对于治疗目的，文献中缺乏共识。不同的群体 已经能够成功地针对骨骼肌或中枢神经系统进行治疗 神经系统采用不同的SBMA动物模型。因此，有必要对SBMA进行研究 在人类背景中，受影响的细胞类型，骨骼肌和运动神经元上的AR突变。 因此，申请人海伦·C·米兰达博士建议结合她在以下领域的丰富经验 干细胞生物学和运动神经元疾病建模成为功能基因组学的导师，以测试 假设在SBMA中AR转录网络是组织特异性的。这个项目将检验这一假设。 通过结合从SBMA和等基因产生的AR基因组占有率和基因表达数据集 控制IPSC衍生的骨骼肌和运动神经元。这项工作将增进对 人突变型AR在SBMA发病中的分子机制及IPSC的应用价值 骨骼肌和运动神经元在体外研究中发展SBMA的工具。米兰达博士是一位新的助理 凯斯西储大学遗传学系和神经科学系教授 大学。她将把75%的时间投入到这个奖项下的研究中，并将用 基因组和转录组分析的教学培训。此培训将包括1)部门和 大学课程，2)研讨会和期刊俱乐部，3)负责任地开展研究课程，4)国家 和国际会议。米兰达医生将由安东尼·温肖-鲍里斯医生和安医生指导 凯斯西储大学的哈里斯。这些老牌科学家都是茎干领域的知名专家。 细胞生物学和功能基因组学。米兰达博士会见了她的每一位导师，讨论了这个项目，并 在此期间，将继续定期与他们会面(在导师信中指定) 获奖。预计她将以通讯或共同通讯作者的身份制作手稿，并 在这个奖项的过程中竞争R-Level奖助金。该项目将整合米兰达博士目前的 具有额外培训的专业知识，以开发全面的、独立的研究计划。