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

基本信息

批准号：
10444917
负责人：
Pei-Ciao Tang
金额：
$ 14.58万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
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/10444917
关键词：
3-Dimensional Acoustic Nerve Address Atlases Automobile Driving Cells Cephalic Congenital Disorders Data Databases Defect Derivation procedure Development Developmental Biology Disease Ectoderm Cell Embryo Epithelial 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 base cell type combat congenital hearing loss deafness experimental study gene expression database gene regulatory network gene therapy human stem cells improved in vivo inner ear development inner ear diseases novel strategies organ growth placodal ectoderm regenerative tissue single-cell RNA sequencing spatiotemporal transcriptome transcriptomics

项目摘要

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.

项目概要/摘要大多数先天性听力损失病例是感音神经性的（SNHL），其中至少一半与遗传缺陷。不幸的是，内耳疾病的再生医学和基因治疗已普遍由于无法在大量人类身上进行合乎道德的实验，尚未在人类患者中实现以科学严谨的方式培育胚胎。这也意味着整体上存在关键知识差距了解人类感觉器官的早期发育。就内耳而言，众所周知细胞命运对耳基板的正确承诺对于内耳的形成至关重要。两个感觉上皮，其中毛细胞位于，前庭蜗神经源自耳囊，内陷来自耳基板；毛细胞和/或前庭蜗神经缺陷会导致 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，耳基板源自哺乳动物胚胎（更不用说人类胚胎）。本研究目的 1) 检查 WNT 信号通路在早期耳发育中的作用，重点关注 PPE，2) 确定 PPE 中的细胞命运决定如何最终导致人类内耳耳系谱系的起源类器官。在目标 1 中，将检查 WNT 信号传导对耳发育的影响，目标是确定最大化内耳类器官诱导的 WNT 最佳水平。单细胞 RNA 测序（scRNA-seq）对人类内耳类器官系统的分析将揭示早期耳谱系的进展和 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创建经过最佳水平的 WNT 调制处理的。将这些空间数据与来自 Aim 1（人类内耳类器官早期耳谱系的细胞图谱）的时间 scRNA-seq 数据将被已确立的。然后，这些时空转录组数据将用于创建早期的遗传蓝图。人类耳发育，重点是对细胞命运至关重要的基因调控网络的特征正确的颅感觉器官发育的承诺。这些数据将 1) 推动人类进步发育生物学，2) 生成一个分子数据库，用于了解相关先天性疾病外胚层衍生物（包括内耳），以及3）再生医学和基因治疗的进展用于内耳疾病。