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.

项目摘要 Barrett的食道和食管中的小细胞癌的特征是鳞状细胞的丧失 同时获得其他细胞命运。这些疾病的有效的治疗方案部分受到限制 由于对驱动初始细胞脂肪变化的分子机制的了解不完全理解。这 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措施 神经内分泌细胞承诺的食管基碱细胞。 我们和其他人已经证明，p63的损失导致分层鳞状上皮的一起 在发展中的食道。但是，p63如何调节食管祖细胞的细胞脂肪测定 令人惊讶的是，我们发现p63缺失会导致广泛的神经内分泌 食道的分化。此外，在小细胞癌中也观察到p63表达的丧失 食道。抑制关键表观遗传沉默因子EZH2重新激活p63在 神经内分泌细胞，这是通过获得鳞状细胞命运来完成的。因此，我们假设 表观遗传调节的p63表达在促进鳞状细胞的同时抑制神经内分泌 食道中的分化计划。制定了三个特定目标以检验假设。目标1意志 测试p63在细胞脂肪确定中的作用是否是同类特异性的，并且在人类中构成 食管祖细胞（EPC）。我们将结合小鼠遗传模型和人类多能干细胞 差异化以解决这个目标。 AIM 2将确定P63介导的转录程序 EPC的规范。我们将使用包括Chip-Seq和ATAC-Seq在内的多种表观基因组分析来确定 p63在全基因组水平上的下游靶标。我们还将使用生物化学测定测试候选基因 在细胞命运过渡中实施，该过渡可能受到p63的可能结合并在功能上调节。 AIM 3将确定 p63沉默对神经内分泌承诺的机制和影响。在此目标中，我们将调查如何 p63表达在异常细胞脂肪承诺过程中通过表观遗传调节机制调节，因此 确定恢复鳞状分化计划的策略。结合我们关于鼠标的联合专业知识 遗传学，细胞生物学，染色质生物学和表观基因组学，这项工作将为您提供新的机械见解 食管细胞命运调节并提供治疗神经内分泌病变的新方法。