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

• 批准号: 8255148
• 负责人: Stefan Heller
• 金额: $ 54.56万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 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/8255148
• 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; embryonic stem cell; genome wide association study; hearing impairment; human embryonic stem cell; induced pluripotent stem cell; insight; mutant; novel; novel strategies; research study; response; treatment strategy

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. PUBLIC HEALTH RELEVANCE: In this grant application it is proposed to generate in the culture dish human inner ear cells. These cells will be derived from human embryonic stem cells and from induced pluripotent stem cells generated from a small skin biopsy taken from hearing loss patients. This novel strategy will establish an indirect form of 'inner ear biopsy' to allow interrogation of the inner ear cellular phenotypes in human patients with a variety of different types of hearing loss. The focus of the grant proposal is on sensory hair cell-like cells and on inner ear supporting cell-like cells. These cell types will be characterized in detail and the pathological changes associated with specific genetic mutations will be measured at the cellular, cell-physiological, and molecular level. Potential ways for therapy will be explored directly on patient-derived cells. A second focus lies on the identification of genetic circumstances that modulate the severity of a common form of human hereditary sensorineural hearing loss. Successful completion of the proposed research will result in an unprecedented novel approach to assess human inner ear dysfunction, diagnosis, and for testing of novel treatment strategies directly on patient-derived cells.

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。 公共卫生相关性：在此赠款应用中，提议在人类内耳细胞中产生。这些细胞将来自人类胚胎干细胞，以及由从听力损失患者中进行的小型皮肤活检产生的诱导多能干细胞。这种新颖的策略将建立一种“内耳活检”的间接形式，以允许对具有多种不同类型的听力损失的人类患者进行内耳细胞表型询问。赠款提案的重点是感觉毛细胞样细胞和支撑细胞样细胞的内耳。这些细胞类型将详细表征，并且将在细胞，细胞生理学和分子水平上测量与特定遗传突变相关的病理变化。潜在的治疗方法将直接在患者衍生的细胞上探索。第二个重点在于识别遗传环境，这些遗传环境调节了人类遗传性感觉性听力损失的常见形式的严重性。成功完成拟议的研究将导致一种史无前例的新方法来评估人类内耳功能障碍，诊断和直接在患者衍生细胞上测试新型治疗策略。