Cell fate determination in the early development of the otic lineage in human inner ear organoids

人内耳类器官耳谱系早期发育中的细胞命运决定

• 批准号: 10645166
• 负责人: Pei-Ciao Tang
• 金额: $ 15.34万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-05 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10645166
• 关键词: 3-Dimensional; Acoustic Nerve; Address; Atlases; Automobile Driving; Cell Aggregation; Cells; Cephalic; Congenital Disorders; Data; Databases; Defect; Derivation procedure; Development; Developmental Biology; Disease; Ectoderm Cell; Embryo; Epithelium; Ethics; Fibroblast Growth Factor; Functional disorder; Future; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genetic; Genetic Diseases; Genetic study; Goals; Hair Cells; Health; Human; In Vitro; Knowledge; Labyrinth; Lead; Maps; Mediating; Modeling; Molecular; Mus; Mutation; Natural regeneration; Neurons; Organ; Organoids; Otic Placodes; Otic Vesicle; Patients; Pattern; Physiology; Play; Positioning Attribute; Process; Productivity; Protocols documentation; Regenerative Medicine; Resolution; Role; Sampling; Sensorineural Hearing Loss; Sensory; Skin; System; Testing; Time; Variant; WNT Signaling Pathway; cell type; combat; congenital hearing loss; deafness; experimental study; gene expression database; gene regulatory network; gene therapy; genetic association; human stem cells; improved; in vivo; inner ear development; inner ear diseases; novel strategies; organ growth; placodal ectoderm; regenerative tissue; single-cell RNA sequencing; spatial integration; spatiotemporal; transcriptome; transcriptomic profiling; transcriptomics

PROJECT SUMMARY/ABSTRACT The majority of congenital hearing loss cases are sensorineural (SNHL), and at least half are associated with genetic defects. Unfortunately, regenerative medicine and gene therapy for inner ear diseases have generally not been realized in human patients due to the inability to ethically experiment on large numbers of human embryos in a scientifically rigorous manner. This also means that key knowledge gaps exist in the overall understanding of the early development of human sensory organs. In the case of the inner ear, it is known that proper cell fate commitment to the otic placode is essential for inner ear formation. Both sensory epithelia, where hair cells are located, and the vestibulocochlear nerve are derived from the otic vesicle, which is invaginated from the otic placode; defects in hair cells and/or the vestibulocochlear nerve result in SNHL. It is difficult to study early otic development, specifically during the induction and patterning of the pre-placodal ectodermal (PPE), where otic placodes are derived from, in mammalian embryos (not to mention human embryos). This study aims to 1) examine the role of WNT signaling pathways in early otic development, with a focus on the PPE, and 2) determine how cell fate decisions in the PPE culminate in the origin of the otic lineage in human inner ear organoids. In Aim 1, the effects of WNT signaling on otic development will be examined, with the goal of identifying the optimal level of WNT that maximizes inner ear organoid induction. Single-cell RNA sequencing (scRNA-seq) analyses in the human inner ear organoid system will reveal the early otic lineage progression and mechanisms underlying the processes mediated by WNT. In Aim 2, spatial gene expression profiles of organoids treated with the optimal level of WNT modulation will be created. Upon integrating these spatial data with the temporal scRNA-seq data from Aim 1, a cell atlas of the early otic lineage in human inner ear organoids will be established. These spatio-temporal transcriptomic data will then be used to create a genetic blueprint for early human otic development, with a focus on characterizing the gene regulatory networks that are critical to cell fate commitments underlying proper cranial sensory organ development. Such data will 1) advance human developmental biology, 2) generate a molecular database for understanding congenital disorders associated with ectodermal derivatives (including the inner ear), and 3) progress regenerative medicine and gene therapy for inner ear diseases.

项目总结/摘要 大多数先天性听力损失病例是感音神经性（SNHL），至少有一半与 遗传缺陷不幸的是，内耳疾病的再生医学和基因治疗通常 由于无法在大量人类患者身上进行伦理实验， 以科学严谨的方式培养胚胎。这也意味着，总体而言， 了解人类感觉器官的早期发育。在内耳的情况下，众所周知， 对耳基板的适当细胞命运定型对于内耳形成是必需的。两个感觉上皮， 毛细胞定位，前庭蜗神经来自内陷的耳泡 来自耳基板;毛细胞和/或前庭耳蜗神经的缺陷导致SNHL。学习很难 早期耳发育，特别是在前基板外胚层（PPE）的诱导和形成过程中， 在哺乳动物胚胎（更不用说人类胚胎）中，耳基板来自哪里。本研究旨在 1）检查WNT信号通路在早期耳部发育中的作用，重点是PPE，以及2） 确定PPE中的细胞命运决定如何最终导致人类内耳中耳谱系的起源 类有机体在目标1中，将检查WNT信号传导对耳发育的影响，目标是 鉴定使内耳类器官诱导最大化的WNT的最佳水平。单细胞RNA测序 人内耳类器官系统中的scRNA-seq分析将揭示早期耳谱系进展， WNT介导的过程的潜在机制。在目标2中，类器官的空间基因表达谱 将产生用最佳水平的WNT调制处理的。在将这些空间数据与 来自Aim 1的时间scRNA-seq数据，人类内耳类器官中早期耳谱系的细胞图谱， 确立了习这些时空转录组学数据将用于创建一个遗传蓝图， 人类耳部发育，重点是表征对细胞命运至关重要的基因调控网络 大脑感觉器官正常发育的基础这些数据将1）促进人类 发育生物学，2）生成分子数据库，用于了解与发育相关的先天性疾病， 与外胚层衍生物（包括内耳），和3）发展再生医学和基因治疗 治疗内耳疾病