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连锁的脊柱和鳞茎肌肉萎缩（也称为SBMA或肯尼迪氏病）是罕见的 因运动神经元较低而导致成人发作的近端肌肉无力的神经肌肉疾病 退化。 SBMA患者表现出雄激素不敏感的迹象，包括妇科，生育力降低， 和有牙性萎缩。 SBMA是由雄激素中的CAG多谷氨酰胺（PolyQ）重复膨胀引起的 接收器（AR）基因，是九个CAG-POLYQ重复疾病的家族的成员，其中包括 亨廷顿氏病。几十年来，对神经系统疾病的基础的研究集中在 神经元功能障碍对疾病发病机理的贡献。但是，在过去的十年中， 运动神经元疾病领域的越来越多的证据挑战了主要的以神经为中心的理论 许多神经系统疾病的病因。最近，我们发现对照电机中的细胞死亡增加 与对照相比 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。但是，出于治疗目的，文献中缺乏共识。不同的组 能够证明针对骨骼肌或中央的成功治疗方法 神经系统使用不同的SBMA动物模型。因此，需要研究SBMA 在人类背景下，受影响的细胞类型，骨骼肌和运动神经元的AR突变。 因此，申请人海伦·C·米兰达（Helen C. Miranda）博士提议将她的丰富经验结合在一起 干细胞生物学和运动神经元疾病建模，以实现功能基因组学的心态，以测试 假设AR转录网络在SBMA中是组织特异性的。该项目将检验此假设 通过结合AR基因组广泛的占用率和基因表达数据集，由SBMA和等源性产生 控制IPSC衍生的骨骼肌和运动神经元。这项工作将促进对 人突变体AR的分子机制可用于SBMA发病机理并评估IPSC衍生的效用 骨骼肌和运动神经元工具，以发展SBMA体外研究。米兰达博士是新助理 Case Western Reserve的遗传学系和神经科学系教授 大学。她将花75％的时间根据该奖项进行研究，并将她的研究补充 基因组和转录组分析中的教学训练。该培训将完成1）部门和 大学课程，2）半手和期刊俱乐部3）负责任的研究课程和4）国家 和国际会议。 Miranda博士将由Anthony Wynshaw-Boris博士和Ann博士考虑 哈里斯（Case Western Reserve University）的哈里斯（Harris）。这些成熟的科学家都是STEM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，讨论这个项目， 在此过程中，将继续定期与他们会面 奖。预计她将产生手稿作为相应或相应的作者，并成为 在本奖项期间，R级赠款的竞争力。该项目将整合米兰达博士的当前 通过额外培训的专业知识，以制定全面的独立研究计划。