Cell Fate Regulation in Esophageal Progenitor Cells

食管祖细胞的细胞命运调控

• 批准号: 10576408
• 负责人: Chao Lu
• 金额: $ 58.69万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10576408
• 关键词: ASCL1 gene; ATAC-seq; Address; Adult; Automobile Driving; Barrett Esophagus; Binding; Binding Sites; Biochemistry; Biological Assay; Biology; Candidate Disease Gene; Cell Differentiation process; Cell Fate Control; Cell Line; Cells; Cellular biology; ChIP-seq; Chromatin; Complex; Data; Data Set; Disease; EZH2 gene; Embryo; Epigenetic Process; Epithelium; Esophageal Diseases; Esophageal carcinoma; Esophagus; Genes; Genetic; Genetic Models; Genetic Transcription; Genome; Germ Cells; Goals; High-Throughput Nucleotide Sequencing; Human; Joints; Knockout Mice; Knowledge; Lesion; Mediating; Modeling; Molecular; Mus; Neuroendocrine Cell; Neuroendocrine Tumors; Neurosecretory Systems; Pathogenesis; Play; Polycomb; Promoter Regions; Protein Isoforms; Role; Small Cell Carcinoma; Specific qualifier value; Squamous Cell; Squamous Differentiation; Stratified Squamous Epithelium; Surveys; Testing; Transposase; Work; derepression; effective therapy; epigenetic regulation; epigenetic silencing; epigenomics; genome-wide; human pluripotent stem cell; inhibitor; insight; insulinoma associated 1; member; mouse genetics; mouse model; neuroendocrine differentiation; new therapeutic target; novel; novel strategies; postnatal; programs; stem cell differentiation; stem cells; transcription factor; transcriptome sequencing

Project Summary Barrett’s esophagus and small cell carcinoma in the esophagus are characterized by the loss of squamous cell identity while gaining other cell fates. Thus far effective treatment options for these diseases are limited in part due to an incomplete understanding of the molecular mechanisms driving the initial cell fate changes. This application will address how the transcription factor p63 directs the squamous differentiation program and safeguards esophageal basal progenitor cells from neuroendocrine cell commitment. We and others have demonstrated that loss of p63 leads to the abrogation of the stratified squamous epithelium in the developing esophagus. However, how p63 regulates cell fate determination of esophageal progenitor cells remains largely unknown. Surprisingly, we found that p63 deletion results in extensive neuroendocrine differentiation in the esophagus. Moreover, loss of p63 expression was also observed in small cell carcinoma of the esophagus. Inhibition of the key epigenetic silencing factor EZH2 re-activates p63 expression in neuroendocrine cells, which is accompanied by the acquisition of squamous cell fate. We therefore hypothesize that epigenetically regulated p63 expression suppresses neuroendocrine while promoting the squamous cell differentiation program in the esophagus. Three specific aims are formulated to test the hypothesis. Aim 1 will test whether the role played by p63 in cell fate determination is isoform-specific and conserved in human esophageal progenitor cells (EPCs). We will combine mouse genetic models and human pluripotent stem cell differentiation to address this Aim. Aim 2 will determine p63-mediated transcriptional program during the specification of EPCs. We will use multiple epigenomic assays including ChIP-seq and ATAC-seq to determine p63 downstream targets at the genome-wide level. We will also use biochemistry assays to test candidate genes implicated in cell fate transition that are potentially bound and functionally regulated by p63. Aim 3 will determine the mechanism and impact of p63 silencing on neuroendocrine commitment. In this Aim we will investigate how p63 expression is modulated by epigenetic regulatory mechanisms during aberrant cell fate commitment, thus identifying strategies to restore the squamous differentiation program. Combining our joint expertise on mouse genetics, cell biology, chromatin biology and epigenomics, this work will provide novel mechanistic insights into esophageal cell fate regulation and offer new approaches to treat neuroendocrine lesions.

项目概要 巴雷特食管和食管小细胞癌的特点是鳞状细胞丢失 身份，同时获得其他细胞的命运。迄今为止，这些疾病的有效治疗方案在一定程度上是有限的 由于对驱动初始细胞命运变化的分子机制的不完全理解。这 该应用程序将解决转录因子 p63 如何指导鳞状分化程序和保障 食管基底祖细胞由神经内分泌细胞定型。 我们和其他人已经证明 p63 的缺失会导致复层鳞状上皮的消失 在发育中的食道中。然而，p63如何调节食管祖细胞的细胞命运决定 仍然很大程度上未知。令人惊讶的是，我们发现 p63 缺失导致广泛的神经内分泌 食管的分化。此外，在小细胞癌中也观察到 p63 表达缺失。 食道。抑制关键表观遗传沉默因子 EZH2 重新激活 p63 表达 神经内分泌细胞的命运是伴随着鳞状细胞的获得而发生的。因此我们假设 表观遗传调控的 p63 表达抑制神经内分泌，同时促进鳞状细胞生长 食管中的分化程序。制定了三个具体目标来检验该假设。目标1将 测试 p63 在细胞命运决定中所发挥的作用在人类中是否具有异构体特异性和保守性 食管祖细胞（EPC）。我们将把小鼠遗传模型和人类多能干细胞结合起来 差异化来解决这一目标。目标 2 将确定 p63 介导的转录程序 EPC 规范。我们将使用包括 ChIP-seq 和 ATAC-seq 在内的多种表观基因组检测来确定 p63 下游目标为全基因组水平。我们还将使用生物化学检测来测试候选基因 参与细胞命运转变，可能受 p63 结合并进行功能调节。目标 3 将决定 p63沉默对神经内分泌承诺的机制和影响。在此目标中，我们将研究如何 p63 表达在异常细胞命运承诺期间受到表观遗传调控机制的调节，因此 确定恢复鳞状分化程序的策略。结合我们在鼠标方面的联合专业知识 遗传学、细胞生物学、染色质生物学和表观基因组学，这项工作将为 食管细胞命运调控并为治疗神经内分泌病变提供新方法。