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

• 批准号: 10615234
• 负责人: Vittorio Sebastiano
• 金额: $ 39.35万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 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/10615234
• 关键词: 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 Deletion; Gene Expression; Generations; Genes; Genetic; Genetic Screening; Genetic Transcription; Heart; Heterozygote; Histones; Human; In Vitro; Intervention; Knowledge; Maps; Mesoderm; Methods; Modeling; Molecular; Mus; Muscle; Mutation; Neural Crest Cell; Nucleic Acid Regulatory Sequences; Parathyroid gland; Pathway interactions; Patients; Pattern; Pharyngeal Apparatus; Pharyngeal structure; Phase; Phenotype; Physiological; Play; Population; Qi; Regulation; Role; Signal Transduction; Structure; Syndrome; System; Testing; Thymus Gland; Tissues; Validation; Work; 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; pharmacologic; pharyngeal patterning; prevent; reconstruction; response; stem cell biology; tool; transcription factor; transcriptome; transcriptome sequencing; whole genome

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.

项目摘要 22Q11.2删除综合征（22q11.2ds）是最常见的微骨骼综合征，是由 在22号染色体上的半合子损失为0.7-3 Mb的DNA，并导致临床表型的星座。 核心表型源于咽部设备的破坏。特别受影响 是第二心地依赖的心脏结构，伟大的血管，甲状旁腺，胸腺和较低 颅面和面部肌肉。尽管大约可以删除大约50个基因，但它是单倍的不足。 转录因子TBX1概括了与22q11.2ds相关的大多数关键表型。 遗传和发育小鼠研究已经确定，TBX1对于典型的发展至关重要 咽内胚层是胸腺开发，甲状旁腺的瞬时解剖结构， 和第四咽弓动脉。尽管有这种中心作用，但对于分子而言，知之甚少 TBX1在咽内胚层中起作用的机制。虽然少数研究试图 为了研究TBX1在人类细胞中的作用，它们所进行的细胞类型并不代表 TBX1起关键作用的适当发展阶段。迄今为止，努力整合 尚未尝试进入咽内胚层或第四个咽弓动脉网络中的所有关键基因，尚未尝试过， 特别是在人类细胞中。 这种R01利用了一个体外模型的最新发展，该模型忠实地模仿了形成和发展 人类咽部内胚层的作品，从而提供了一个前所未有的机会来逗弄 TBX1在其生理背景下。具体而言，该模型将用于识别转录目标和 TBX1的伴侣（AIM1），研究TBX1作为人咽内胚层的表观遗传调节剂的作用 （AIM2），并机械地研究了TBX1基因座的新发现的定义调节区（AIM3）。 总体假设是，TBX1是基因调节网络的中心，至关重要 咽内胚层的形成和成熟和周围中胚层的形态图案 和神经rest细胞。 预计拟议的工作将根据TBX1单倍宽度和 确定可以通过药理学干预来营救的途径。表观遗传学的解剖 负责咽内胚层形成的分子机械将有助于告知 生成22q11.2ds的细胞疗法。