Regulating transcription of the key pulmonary neuroendocrine lineage driver ASCL1

调节关键肺神经内分泌谱系驱动因子 ASCL1 的转录

• 批准号: 10271277
• 负责人: Jane E Johnson
• 金额: $ 20.5万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-25 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10271277
• 关键词: 3-Dimensional; ASCL1 gene; ATAC-seq; Adult; Automobile Driving; Cell Count; Cell Culture Techniques; Cell Differentiation process; Cell Line; Cell Lineage; Cells; Child; Chromatin; Chronic Obstructive Airway Disease; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Development; Developmental Gene; Developmental Process; Disease; Distal; Distant; Embryo; Enhancers; Epithelial; Epithelial Cells; Future; Gene Expression; Gene Silencing; Genes; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; Homeostasis; Human; Human Cell Line; Knowledge; Learning; Lung; Lung Neuroendocrine Neoplasm; Lung diseases; Malignant Neoplasms; Modeling; Molecular; Molecular Conformation; Neuroendocrine Cell; Neurosecretory Systems; Oncogenes; Play; Population; Protocols documentation; RB1 gene; Regulation; Regulatory Element; Reporting; Role; Signal Transduction; Specific qualifier value; Sudden infant death syndrome; System; Technology; Testing; Transcriptional Regulation; Untranslated RNA; bronchial epithelium; cell type; design; developmental disease; induced pluripotent stem cell; insight; interest; lung cancer cell; lung development; lung injury; lung small cell carcinoma; neuroendocrine cancer; neuroendocrine differentiation; progenitor; stem cell genes; success; transcription factor

The bHLH transcription factor ASCL1 (HASH1/MASH1) is essential for specification of the neuroendocrine (NE) lineage in the lung. NE cells are rare in pulmonary epithelium but they are thought to play diverse roles during lung development and homeostasis. Increases in NE cell number have been documented in many lung diseases in children and adults. ASCL1 function is balanced with NOTCH signaling to control progenitor proliferation and NE differentiation versus other cell types in the lung. Consistent with the role of ASCL1 as a lineage-specifying factor and in expanding progenitor populations, ASCL1 has been found to be a lineage-dependent oncogene in pulmonary NE tumors as well. With these important functions attributed to ASCL1, and its requirement for controlled spatial and temporal expression during development and in conditions of lung damage, it is surprising how little is known about regulation of ASCL1 gene transcription. This gap in knowledge reflects past technical challenges in identifying and manipulating cis-regulatory elements (REs) found at large distances from the gene of interest, in addition to challenges of poor culture systems reflecting this NE population and the low fraction of NE cells in embryonic and adult lungs. REs functioning at long-distances to control key developmental genes are being discovered using advances in technologies that can interrogate and manipulate the spatial genome. Here we will exploit these technologies to gain much needed insights into transcriptional control of ASCL1 using newly reported protocols for generating pulmonary NE cells from human iPSCs and leveraging cell culture models of neuroendocrine cancer that express high levels of ASCL1. Each model has a particular strength that allows unique aspects of ASCL1 regulation to be uncovered. Aims include identifying and testing functions of long-range REs controlling ASCL1 during NE differentiation in the two models. Success in these aims will provide functional non-coding regulatory sequences controlling ASCL1 expression, and presumably, the NE lineage in lung development and homeostasis. This is important for future projects to identify molecular components of the signaling complexes working through these REs to reach the goal of providing an understanding of how a key lineage defining transcriptional regulator is controlled during development and disease.

BHLH转录因子ASCL1（HASH1/MASH1）对于规范 肺中神经内分泌（NE）谱系。 NE细胞在肺上皮中很少见，但它们是 被认为在肺部发育和体内平衡期间扮演着各种角色。 NE细胞增加 在儿童和成人的许多肺部疾病中已经记录了数量。 ASCL1函数是 与缺口信号平衡以控制祖细胞增殖和NE分化与 肺中的其他细胞类型。与ASCL1作为谱系指定因素的作用一致，并且 在扩大的祖先种群中，已发现ASCL1是谱系依赖性的 肺NE肿瘤中的癌基因。这些重要功能归因于ASCL1， 及其在开发过程中以及在 肺损伤的条件，令人惊讶的是，关于ASCL1基因的调节知之甚少 转录。知识的差距反映了过去的技术挑战和 操纵与感兴趣基因的距离很大的操纵顺式调节元件（RES） 补充反映该国内国家人群和低部分的贫困文化系统的挑战 胚胎和成年肺中的NE细胞。 RES在长距离运行以控制密钥 使用可以审问的技术的进步发现了发展基因 并操纵空间基因组。在这里，我们将利用这些技术来获得很多 需要使用新报告的协议对ASCL1进行转录控制的见解 从人IPSC产生肺NE细胞，并利用 表达高水平ASCL1的神经内分泌癌。每个模型都有特殊的强度 这允许发现ASCL1调节的独特方面。目的包括识别和 在两者中的NE分化过程中控制ASCL1的远程RES的测试功能 型号。这些目标的成功将提供功能性的非编码调节序列 控制ASCL1的表达，并且大概是肺发育中的NE谱系和 稳态。这对于将来的项目确定分子成分很重要 通过这些RES运行的信号复合物，以实现提供理解的目标 关于如何在开发过程中控制定义转录调节器的关键谱系和 疾病。