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: 10400920
• 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/10400920
• 关键词: AR gene; Address; Adult; Affect; Androgen Receptor; Androgens; Animal Model; Atrophic; Award; Biological Assay; CRISPR interference; Cell Death; Cells; 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; Neuraxis; 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; deep sequencing; defined contribution; differential expression; experience; functional genomics; 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.

概括 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 神经肌肉疾病，其特征是成人发病的下运动神经元引起的近端肌肉无力 SBMA 患者表现出雄激素不敏感的迹象，包括男性乳房发育、生育能力下降、 和 SBMA 是由雄激素中的 CAG-聚谷氨酰胺 (polyQ) 重复扩张引起的。 受体 (AR) 基因，是九种 CAG-polyQ 重复疾病家族的成员之一，其中包括 几十年来，对神经系统疾病基础的研究主要集中在亨廷顿舞蹈症。 神经元功能障碍对疾病发病机制的贡献然而，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理论 最近，我们发现控制运动细胞死亡增加。 与对照相比，接受来自 SBMA iPSC 衍生骨骼肌的条件培养基的神经元 这一发现强调了肌肉毒性在 SBMA 疾病中的重要性。 然而，对于治疗目的，不同的研究小组缺乏共识。 已经能够证明针对骨骼肌或中枢的成功治疗 神经系统使用不同的SBMA动物模型进行检查，有必要对SBMA进行研究。 在人类背景下，受影响的细胞类型、骨骼肌和运动神经元发生 AR 突变。 因此，申请人 Helen C. Miranda 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到功能基因组学的指导，以测试 AR 转录网络在 SBMA 中具有组织特异性的假设本项目将检验这一假设。 通过结合 SBMA 和 isogenic 生成的 AR 全基因组占用和基因表达数据集 控制 iPSC 衍生的骨骼肌和运动神经元。这项工作将增进对 iPSC 衍生的骨骼肌和运动神经元的理解。 人类突变体 AR 对 SBMA 发病机制的分子机制并评估 iPSC 衍生的效用 Miranda 博士是开发 SBMA 体外研究骨骼肌和运动神经元工具的新助理。 凯斯西储大学遗传学系和神经科学系教授 大学期间，她将投入 75% 的时间进行该奖项的研究，并以其他方式补充她的研究。 基因组和转录组分析方面的教学培训将包括 1) 部门和部门。 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，以及 4) 国家级课程 米兰达博士将由安东尼·温肖-鲍里斯博士和安博士指导。 凯斯西储大学的哈里斯这些知名科学家都是干细胞领域的著名专家。 细胞生物学和功能基因组学米兰达博士与她的每位导师会面讨论了这个项目并 在此期间，将继续定期与他们会面（在指导信中指定） 她预计将作为通讯作者或共同通讯作者撰写手稿并获得该奖项。 在该奖项期间，该项目将整合米兰达博士目前的研究成果，以获取 R 级资助。 专业知识和额外培训，以开发全面的、独立的研究计划。