Dissecting the Molecular Role of TBX1 in the Context of Human Pharyngeal Endoderm Development

解析 TBX1 在人咽内胚层发育中的分子作用

基本信息

批准号：
10442889
负责人：
Vittorio Sebastiano
金额：
$ 39.35万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10442889
关键词：
22q11 22q11.2 ATAC-seq Affect Anatomy Animal Model Arteries Binding Biocompatible Materials Biology Branchial arch structure CRISPR interference Cell Therapy Cell model Cells ChIP-seq Chromatin Chromosome 22 Congenital Heart Defects DNA Data Development DiGeorge Syndrome Disease Dissection Embryonic Development Endoderm Endoderm Cell Endothelium Enhancers Ensure Epigenetic Process Face Gene Expression Generations Genes Genetic Genetic Screening Genetic Transcription Heart Histones Human In Vitro Intervention Knowledge Maps Mesoderm Methods Modeling Molecular Molecular Genetics Mus Muscle Mutation Neural Crest Cell Nucleic Acid Regulatory Sequences Parathyroid gland Pathway interactions Patients Pattern Pharmacology Pharyngeal Apparatus Pharyngeal structure Phase Phenotype Physiological Play Population Qi Regulation Role Signal Transduction Structure Syndrome System Testing Thymus Gland Tissues Validation Work base cell type clinical phenotype craniofacial developmental disease dosage epigenetic regulation experimental study gene regulatory network genetic variant genome sequencing human embryonic stem cell human model human tissue in vitro Model in vivo insight microdeletion multidisciplinary new technology novel pharyngeal patterning prevent reconstruction response stem cell biology tool transcription factor transcriptome transcriptome sequencing whole genome

项目摘要

PROJECT SUMMARY 22q11.2 Deletion Syndrome (22q11.2DS), the most common of the microdeletion syndromes, is caused by hemizygous loss of 0.7-3 Mb of DNA on chromosome 22 and results in a constellation of clinical phenotypes. The core phenotype originates from disrupted development of the pharyngeal apparatus. Particularly affected are the second heart field-dependent heart structures, great vessels, parathyroids, thymus, and lower craniofacial and face muscles. Although approximately 50 genes may be deleted, it is the haploinsufficiency of the transcription factor TBX1 that recapitulates most of the critical phenotype associated with 22q11.2DS. Genetic and developmental mouse studies have established that TBX1 is critical for typical development of the pharyngeal endoderm, a transient anatomical structure necessary for development of the thymus, parathyroids, and 4th pharyngeal arch arteries. Despite this central role, very little is known regarding the molecular mechanisms by which TBX1 functions in the pharyngeal endoderm. While a handful of studies have attempted to study the role of TBX1 in human cells, the cell types they have been conducted in are not representative of the appropriate developmental stage where and when TBX1 plays its critical role. To date, an effort to integrate all the critical genes into a pharyngeal endoderm or 4th pharyngeal arch arteries network has not been attempted, particularly in human cells. This R01 leverages recent development of an in vitro model which faithfully mimics the formation and progression of human pharyngeal endoderm, thereby providing an unprecedented opportunity to tease out the functions of TBX1 in its physiological context. Specifically, this model will be used to identify the transcriptional targets and partners of TBX1 (Aim1), investigate the role of TBX1 as epigenetic regulator of the human pharyngeal endoderm (Aim2), and mechanistically investigate newly discovered putative regulatory regions of the TBX1 locus (Aim3). The overarching hypothesis is that TBX1 is at the center of a Gene Regulatory Network critical for both the formation and maturation of the pharyngeal endoderm and the morpho-patterning of the surrounding mesoderm and neural crest cells. The proposed work is expected to identify the molecular mechanism at the basis of TBX1 haploinsufficiency and identify pathways which could be rescued through pharmacological intervention. Dissection of the epigenetic and molecular machinery responsible for pharyngeal endoderm formation will be instrumental in informing the generation of cell therapies for 22q11.2DS.

项目概要 22q11.2 缺失综合征 (22q11.2DS) 是最常见的微缺失综合征，由 22 号染色体上 0.7-3 Mb DNA 的半合子丢失并导致一系列临床表型。核心表型源于咽部发育中断。特别受影响是第二心脏场依赖的心脏结构、大血管、甲状旁腺、胸腺和下丘脑颅面和面部肌肉。虽然大约有50个基因可能被删除，但这是单倍体不足转录因子 TBX1 概括了与 22q11.2DS 相关的大部分关键表型。遗传和发育小鼠研究已证实 TBX1 对于小鼠的典型发育至关重要咽内胚层，胸腺、甲状旁腺发育所必需的短暂解剖结构，和第四咽弓动脉。尽管具有这一核心作用，但人们对分子的了解却知之甚少。 TBX1 在咽内胚层中发挥作用的机制。虽然一些研究尝试为了研究 TBX1 在人类细胞中的作用，他们所进行的细胞类型并不代表 TBX1 发挥关键作用的适当发育阶段。迄今为止，努力整合尚未尝试将所有关键基因导入咽内胚层或第四咽弓动脉网络，特别是在人体细胞中。该 R01 利用了最新开发的体外模型，忠实地模拟了形成和进展人类咽内胚层的功能，从而为梳理其功能提供了前所未有的机会 TBX1 的生理背景。具体来说，该模型将用于识别转录目标和 TBX1 (Aim1) 的合作伙伴，研究 TBX1 作为人咽内胚层表观遗传调节因子的作用 (Aim2)，并从机制上研究新发现的 TBX1 基因座的假定调控区域 (Aim3)。总体假设是 TBX1 位于基因调控网络的中心，对于咽内胚层的形成和成熟以及周围中胚层的形态模式和神经嵴细胞。所提出的工作预计将在 TBX1 单倍体不足和确定可以通过药物干预来挽救的途径。表观遗传学的剖析负责咽内胚层形成的分子机制将有助于告知 22q11.2DS 细胞疗法的产生。