How Patterned Mesenchymal-Epithelial interactions Shape Intestinal Crypts

图案化的间充质-上皮相互作用如何塑造肠隐窝

• 批准号: 10571923
• 负责人: Ophir D Klein
• 金额: $ 59.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10571923
• 关键词: Adult; Bioinformatics; Biology; Cell Differentiation process; Cells; Characteristics; Coculture Techniques; Colon; Colon Carcinoma; Colon Injury; Colonic Diseases; Colorectal Cancer; Communication; Development; Differentiation and Growth; Disease; Embryo; Engineering; Epithelial Cells; Epithelium; Event; Fibroblasts; Genetic; Genetic Transcription; Goals; Health; Histologic; Homeostasis; Human; In Vitro; Inflammatory Bowel Diseases; Injury; Knowledge; Maintenance; Maps; Mesenchymal; Mesenchyme; Modeling; Molecular; Morphogenesis; Morphology; Mucous Membrane; Mus; Natural regeneration; Organoids; Pathology; Pattern; Play; Population; Principal Investigator; Productivity; Regulation; Research; Role; Shapes; Signal Transduction; Site; Small Intestines; Structure of intestinal gland; System; Testing; Therapeutic; Undifferentiated; Villus; WNT Signaling Pathway; Work; base; collaborative environment; colonic crypt; comparative; epithelium regeneration; genetic approach; human disease; in vitro Model; in vivo; inhibitor; interest; intestinal crypt; intestinal epithelium; mouse model; progenitor; programs; reconstitution; regenerative; single-cell RNA sequencing; spatiotemporal; stem; stem cells; synthetic biology; transcriptomics

Project Summary/Abstract The colonic epithelium is the site of a host of human diseases, and therefore understanding its development and function is essential for human health. The colon epithelium is separated into morphologically and functionally distinct domains, with proliferative stem cells being housed at the bottom of the crypts of Lieberkühn, whereas differentiated cells populate the epithelium closest to the lumen. The maintenance of epithelial homeostasis in the crypt is dictated in large part by spatially patterned signals from the underlying mesenchyme. The mesenchymal cells involved in adult colonic crypt homeostasis have been of recent interest. However, little is known about the molecular mechanisms underlying colonic crypt formation during development. Our proposal will determine how mesenchymal cell populations and signals interact with the developing epithelium to drive colonic crypt morphogenesis and define stem cell dynamics during development and regeneration. We will accomplish this work through our three aims: First, we will map mesenchymal-epithelial interactions over the course of colonic crypt development using single cell RNAseq and histological approaches. We will then use genetically modified mouse models to investigate the specific role of mesenchymal WNT signals in three stages of colonic crypt development: invagination, elongation, and fission of colonic crypts. Second, we will investigate the potential re-activation of an embryonic program in both the epithelium and mesenchyme following injury induced by DSS treatment. We will test the ability of embryonic or injury-associated mesenchyme to stimulate plasticity, defined as de-differentiation of committed progenitors to a more multipotent state, in the adult epithelium. Third, we will take a synthetic approach to reconstruct patterned mesenchymal signals around colon spheroids, and we will use this new model to test the sufficiency of polarized WNT signals to drive colonic crypt formation in vitro. Overall, our project leverages a combination of in vivo studies, in vitro models, synthetic engineering and bioinformatic approaches. This comprehensive scope is made possible through a highly collaborative environment at UCSF and the productive partnership of the two principal investigators carrying out this project. The findings from this proposal will significantly advance our basic knowledge of colon development and regeneration, highlighting the dynamics of mesenchymal-epithelial crosstalk. This will have implications beyond our fundamental understanding of colon biology and will help pave the way for discovery of therapeutic strategies to treat and modulate colonic disease.

项目概要/摘要 结肠上皮是许多人类疾病的发生部位，因此了解其发展和 功能对人类健康至关重要。结肠上皮从形态上分为形态上皮和功能上皮。 不同的域，增殖干细胞位于 Lieberkühn 隐窝的底部，而 分化的细胞聚集在最接近管腔的上皮细胞中，维持上皮细胞的稳态。 隐窝在很大程度上是由来自底层间充质的空间图案信号决定的。 参与成人结肠隐窝稳态的间充质细胞最近引起了人们的兴趣。 了解发育过程中结肠隐窝形成的分子机制。 将决定间充质细胞群和信号如何与发育中的上皮相互作用 驱动结肠隐窝形态发生并定义干细胞在发育和再生过程中的动力学。 我们将通过三个目标来完成这项工作：首先，我们将绘制间充质-上皮相互作用图谱 然后我们将使用单细胞 RNAseq 和组织学方法进行结肠隐窝发育的过程。 转基因小鼠模型研究间充质WNT信号在三个阶段的具体作用 结肠隐窝发育的过程：结肠隐窝的内陷、伸长和裂变。 损伤后上皮和间质中胚胎程序的潜在重新激活 我们将测试胚胎或损伤相关间充质的刺激能力。 可塑性，定义为在成人中定向祖细胞去分化至更具多能状态 第三，我们将采用合成方法来重建结肠周围的图案化间质信号。 球体，我们将使用这个新模型来测试极化 WNT 信号驱动结肠隐窝的充分性 总的来说，我们的项目结合了体内研究、体外模型、合成。 工程和生物信息方法使这种全面的范围成为可能。 加州大学旧金山分校的协作环境以及两位主要研究人员富有成效的伙伴关系 该项目的研究结果将显着提高我们对结肠发育的基础知识。 和再生，突出间充质-上皮串扰的动态，这将产生影响。 超出我们对结肠生物学的基本理解，并将有助于为治疗方法的发现铺平道路 治疗和调节结肠疾病的策略。