Investigating the Role of Epiregulin During Human Intestinal Development

研究上皮调节蛋白在人类肠道发育过程中的作用

• 批准号: 10677386
• 负责人: Charlie Childs
• 金额: $ 4.75万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10677386
• 关键词: Adult; Animal Model; Biochemical; Biological; Biological Assay; CRISPR/Cas technology; Cell Differentiation process; Cells; Chromatin; Complex; Conceptions; Cues; Data; Development; Disease; Duodenum; Endothelium; Epidermal Growth Factor; Epiregulin; Epithelium; Event; Family member; Gene Expression; Genes; Genomic approach; Genomics; Goals; Growth Factor; Heterogeneity; Homeostasis; Human; Human Biology; Human Development; Immune; Immunofluorescence Immunologic; In Vitro; Intestines; Kidney; Lead; Ligands; Light; Location; MAP Kinase Gene; Maintenance; Mesenchymal; Mesenchyme; Modeling; Molecular; Morphogenesis; Morphology; Mus; Muscularis Mucosa; Natural regeneration; Neurons; Organoids; Outcome; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological; Play; Population; Role; Sampling; Secretory Cell; Signal Transduction; Smooth Muscle; Stomach; Structure; System; Testing; Transplantation; Villus; Western Blotting; Work; Xenograft Model; Xenograft procedure; base; capsule; cell type; experimental study; fetal; human data; human model; in vivo; induced pluripotent stem cell; inhibitor; insight; intestinal epithelium; intestinal homeostasis; loss of function; multiple omics; new growth; novel; organoid transplantation; pharmacologic; self-renewal; single-cell RNA sequencing; stem; stem cell function; stem cell homeostasis; stem cell niche; stem cells; transcription factor

PROJECT ABSTRACT Intestinal Stem Cells (ISCs) are responsible for maintenance and regeneration of the intestinal epithelium, and reside in a domain called the crypt, supported by physical and biochemical cues that make up the stem cell niche. Despite the vital role ISCs play in human intestinal development, homeostasis and disease, the signaling cues that make up the human ISC niche are poorly understood. By analyzing single cell RNA sequencing data from human duodenal samples spanning approximately 7-to-21-week post conception, we determined that Epidermal Growth Factor (EGF), which is commonly used to create an artificial in vitro niche for growing intestinal organoids, was expressed on the differentiated cells of the villus, far from the stem cell domain. We also identified another EGF family member, Epiregulin (EREG), which was highly enriched in the stem cell domain and spatial location confirmed to the crypt. Functional experiments suggest that culturing isolated intestinal epithelium (known as enteroids) in EREG or EGF lead to dramatically different cellular outcomes and spatial organization. Based on this data, the overarching goal of this proposal is to determine the functional role of EREG as a novel intestinal stem cell niche cue. We will test the hypothesis that EREG, and not EGF, is a critical physiologically relevant growth factor for intestinal stem cell homeostasis and plays a key role in human intestinal development. We will test this hypothesis through the following specific aims: 1) Elucidate the mechanism by which EGF and EREG act on human ISCs; 2) Assess the role of EREG in a complex model of the developing human intestine in vitro and following xenotransplantation.

项目摘要 肠干细胞（ISCs）负责肠上皮的维持和再生， 存在于一个称为地穴的区域，由组成干细胞的物理和生物化学线索支持 利基尽管ISCs在人类肠道发育、体内平衡和疾病中起着重要作用， 构成人类ISC小生境的信号传导线索知之甚少。通过分析单细胞RNA 从受孕后约7-21周的人十二指肠样本中获得的测序数据，我们 确定表皮生长因子（EGF），这是常用的创造一个人工体外生态位， 在肠类器官的发育过程中，表达在绒毛的分化细胞上，而不是在干细胞上 域我们还确定了另一个EGF家族成员，表皮调节蛋白（EREG），这是高度富集在表皮生长因子受体。 干细胞结构域和空间位置确认为隐窝。功能实验表明，培养 EREG或EGF中的分离的肠上皮（称为肠上皮）导致细胞分化， 成果和空间组织。根据这些数据，本提案的总体目标是确定 EREG作为一种新的肠道干细胞生态位线索的功能作用。我们将测试EREG， 而不是EGF，是肠道干细胞稳态的关键生理相关生长因子， 在人类肠道发育中的关键作用。我们将通过以下具体目标来检验这一假设：1） 阐明EGF和EREG作用于人ISCs的机制; 2）评估EREG在人ISCs中的作用， 体外发育中的人肠和异种移植后的复杂模型。