Genetically Mediated Hair Cell Degeneration in 3D Inner Ear Organoids

3D 内耳类器官中遗传介导的毛细胞变性

• 批准号: 9892994
• 负责人: Rick F Nelson
• 金额: $ 17.88万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9892994
• 关键词: 3-Dimensional; Advisory Committees; Affect; Antibodies; Binding Proteins; Bioinformatics; Biological Assay; Biological Process; C-terminal; CASP3 gene; CRISPR/Cas technology; Cell Count; Cell Death; Cell physiology; Cells; Cleaved cell; Clustered Regularly Interspaced Short Palindromic Repeats; Cochlea; DNA Sequence Alteration; Data; Development; Disease model; Doctor of Philosophy; Enrollment; Environment; Epithelial; Epithelium; Epitopes; Exhibits; Family; Functional disorder; Gene Expression; Gene Expression Profile; Genes; Genetic; Genomic approach; Genomics; Goals; Hair Cells; Hour; Human; Human Genetics; Immunoprecipitation; In Vitro; Incubators; Indiana; Integral Membrane Protein; International; Iowa; Knockout Mice; Knowledge; Label; Labyrinth; Lead; Maintenance; Mass Spectrum Analysis; Mediating; Mentors; Methods; Modeling; Mus; Mutation; Nature; Organoids; Otolaryngology; Pathway Analysis; Pathway interactions; Patients; Pattern; Postdoctoral Fellow; Potassium Channel; Prevention; Process; Program Development; Protease Domain; Proteins; Proteomics; Research; Research Personnel; Role; Scanning Electron Microscopy; Scientist; Sensory; Sensory Hair; Serine Protease; Structure; System; Technology; Testing; Therapeutic Intervention; Time; Training; Training Programs; Universities; University resources; Vestibular Hair Cells; Work; career; career development; deaf; embryonic stem cell; equilibration disorder; experimental study; extracellular; genome editing; hearing impairment; in vivo; medical schools; member; novel; ototoxicity; preservation; prevent; professor; programs; progressive hearing loss; single-cell RNA sequencing; skills; stem cell biology; stem cells; three dimensional cell culture; three-dimensional modeling; transcriptome; transcriptome sequencing

PROJECT SUMMARY This proposal describes a 5-year training program for the development of a clinician-scientist investigator in stem cell biology and large-scale genomics. The primary goal of this application is to serve as a basis for training Rick Nelson, MD, PhD in the critical skills necessary to become an independent scientific investigator studying inner ear hair cell degeneration. Dr. Nelson is currently an Assistant Professor at Indiana University School of Medicine where he will expand his scientific skills through course work, university resources and mentoring. Hearing loss and vestibular dysfunction affect millions of people worldwide and the common pathophysiologic finding is loss of the mechanosensitive hair cells. Eri Hashino, PhD will serve as Dr. Nelson's primary mentor and train him in stem cell biology. Dr. Hashino is an international leader is stem cell biology and has trained numerous postdoctoral fellows and clinician-scientists. Richard JH Smith, MD at University of Iowa will serve as Dr. Nelson's co-mentor and train him in large-scale genomic approaches and bioinformatics. Dr. Smith is an internationally renowned clinician-scientist and leader in human genetics. Dr. Nelson will also enroll in the Independent Investigator Incubator (I3) program at Indiana University, which provides guidance in scientific and career development. D. Wade Clapp, MD is an internationally recognized clinician-scientist who will serve as a career advisor for Dr. Nelson. His advisory committee will provide broad expertise that spans research planning and career development. In this proposal, the PI will establish a novel model of hair cell degeneration using mouse stem cell-derived 3D cultures containing mechanosensitive hair cells. CRISPR gene editing technology will be used to disrupt the Tmprss3 gene, which is known to cause hair cell degeneration in mice and humans. Next, the PI will use proteomic approaches to establish the protein interactome of the Tmprss3 serine protease in hair cells. Finally, single-cell RNA sequencing (RNA-seq) technology will be used to study global alterations in gene expression that lead to hair cell degeneration. The scientific environment, support from the Department of Otolaryngology at Indiana University and the internationally renowned mentors together make this project feasible. These factors maximize the potential to attain Dr. Nelson's long-term career goal of becoming an independent clinician-investigator and discovering new targets to prevent or delay hair cell degeneration.

项目概要 该提案描述了一项为期 5 年的培训计划，旨在培养临床医生-科学家研究者 干细胞生物学和大规模基因组学。该应用程序的主要目标是作为基础 培训 Rick Nelson 医学博士、哲学博士掌握成为独立科学研究人员所需的关键技能 研究内耳毛细胞变性。尼尔森博士目前是印第安纳大学的助理教授 医学院，他将通过课程作业、大学资源和 指导。听力损失和前庭功能障碍影响着全世界数以百万计的人， 病理生理学发现是机械敏感毛细胞的丧失。 Eri Hashino 博士将担任 Nelson 博士的 干细胞生物学方面的主要导师并对其进行培训。桥野博士是干细胞生物学领域的国际领军人物 并培养了许多博士后研究员和临床科学家。 Richard JH Smith，医学博士，美国大学 爱荷华州将担任尼尔森博士的共同导师，并对他进行大规模基因组方法和生物信息学方面的培训。 史密斯博士是国际知名的临床科学家和人类遗传学领域的领导者。尼尔森博士还将 参加印第安纳大学的独立研究者孵化器 (I3) 项目，该项目提供以下方面的指导： 科学和职业发展。 D. Wade Clapp 医学博士是一位国际公认的临床医生科学家 将担任尼尔森博士的职业顾问。他的咨询委员会将提供广泛的专业知识，涵盖 研究规划和职业发展。在本提案中，PI 将建立一种新型毛细胞模型 使用含有机械敏感毛细胞的小鼠干细胞衍生的 3D 培养物进行变性。基因编辑技术 基因编辑技术将用于破坏 Tmprss3 基因，该基因已知会导致毛细胞 小鼠和人类的退化。接下来，PI 将使用蛋白质组学方法来建立蛋白质 毛细胞中 Tmprss3 丝氨酸蛋白酶的相互作用组。最后，单细胞 RNA 测序 (RNA-seq) 技术将用于研究导致毛细胞退化的基因表达的整体变化。这 科学环境、印第安纳大学耳鼻喉科系和 国际知名导师共同使这个项目变得可行。这些因素最大限度地发挥了潜力 实现尼尔森博士的长期职业目标，即成为一名独立的临床研究人员并发现 预防或延缓毛细胞退化的新目标。