Patient derived sensory hair- and supporting cell-like cells

患者来源的感觉毛发和支持细胞样细胞

基本信息

批准号：
8915331
负责人：
Stefan Heller
金额：
$ 7万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-07 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8915331
关键词：
Animal Model Apical Applications Grants Biopsy Candidate Disease Gene Cells Clinical Complex Genetic Trait Data Development Diagnosis European Exons Fibroblasts Functional disorder Gap Junctions Gene Expression Profile Gene Mutation Genes Genetic Genotype Goals Hair Hair Cells Homozygote Human Human Genetics In Vitro Inherited Labyrinth Measures Mechanical Stimulation Molecular Morphology Motor Mus Mutation Patients Persons Phenotype Physiological Physiology Population Property Proteins Protocols documentation Research Sensorineural Hearing Loss Sensory Hair Severities Skin Stereocilium Supporting Cell Technology Testing Translating Variant Work base cell type clinically relevant cohort deep sequencing embryonic stem cell genome wide association study hearing impairment human embryonic stem cell induced pluripotent stem cell insight mutant novel novel strategies rare variant research study response treatment strategy

项目摘要

DESCRIPTION (provided by applicant): The overriding aim of this grant proposal is to translate technology developed with murine embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) to the clinical setting. More specifically, it is proposed to generate and characterize human inner ear sensory hair cell-like cells and supporting cell-like cells from ESCs and iPSCs. In Aim 1, human ESC-derived hair cell-like cells are being characterized immunocytochemically, morphologically, and functionally. In Aim 2, it is proposed to generate and characterize hair cell-like cells from human iPSCs, specifically from patients carrying mutations in the MYO15A gene. It is expected that the consequences of the mutant MYO15A gene are reflected in the cellular phenotype of hair cell-like cells generated from hearing loss patients. Further proposed is a rescue experiment with wild type MYO15A to restore the cellular phenotype in patient-derived hair cell-like cells. Finally, Aim 3 focuses on characterization of gap junctions in supporting cell-like cells derived from DFNB1 patients homozygous for the common GJB2 35delG mutation. It is proposed to compare cellular properties of 35delG homozygotes with severe-to-profound hearing loss to 35delG homozygotes with mild-to-moderate hearing loss. Measures include gap junction physiology and whole transcriptome analysis, which will be combined with genome wide association studies in a large cohort of DFNB1 patients. The goal of this endeavor is the identification of clinically relevant genetic modifiers, which is a first step toward developing strategies to ameliorate the typical severe phenotype associated with the 35delG/35delG genotype in DFNB1 patients.

描述（由申请人提供）：该赠款提案的覆盖目的是翻译使用鼠类胚胎干细胞（ESC）开发的技术，并诱导多能干细胞（IPSC）到临床环境。更具体地说，提议生成和表征人内耳感觉毛细胞样细胞，并从ESC和IPSC中支撑细胞样细胞。在AIM 1中，人类ESC衍生的毛细胞样细胞在化学上，形态和功能上被表征。在AIM 2中，提议生成和表征来自人IPSC的毛细胞样细胞，尤其是来自Myo15a基因中携带突变的患者。可以预期，突变体Myo15a基因的后果反映在听力损失患者产生的毛细胞样细胞的细胞表型中。进一步提出的是对野生型Myo15a进行的救援实验，以恢复患者衍生的毛细胞样细胞中的细胞表型。最后，AIM 3的重点是在支持源自DFNB1患者的细胞样细胞中表征间隙连接的表征。提议比较35DELG纯合子的细胞性质，其严重的听力损失与35DELG纯合子具有轻度到中度的听力损失。措施包括间隙连接生理学和整个转录组分析，将与大量DFNB1患者组合基因组宽的关联研究结合使用。这项努力的目的是鉴定临床相关的遗传修饰剂，这是制定策略来改善DFNB1患者中与35DELG/35DELG基因型相关的典型严重表型的第一步。