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: 10338737
• 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/10338737
• 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; Epithelial Cells; 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; 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 cell proliferation; 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.

项目摘要/摘要 结肠上皮是许多人类疾病的所在地，因此了解其发展和 功能对于人类健康至关重要。结肠上皮在形态学和功能上分为 不同的域，其中增殖的干细胞位于Lieberkühn地下室的底部，而 分化细胞填充最接近管腔的上皮。维持上皮稳态 地穴在很大程度上取决于来自基础间质的空间图案信号。 与成人结肠隐窝稳态有关的间充质细胞最近引起了人们的关注。但是，几乎没有 关于发育过程中结肠加密形成的分子机制已知。我们的建议 将确定Messedectemal细胞群体和信号如何与发育中的上皮相互作用 在发育和再生过程中驱动结肠隐窝形态发生并定义干细胞动力学。 我们将通过我们的三个目标来完成这项工作：首先，我们将映射间质上皮相互作用 使用单细胞RNASEQ和组织学方法的结肠隐窝发育过程。然后我们将使用 基因修饰的小鼠模型，研究了三个阶段中间充质Wnt信号的特定作用 结肠隐窝的发展：结肠隐窝的内陷，伸长和裂变。第二，我们将调查 损伤后上皮和间质的胚胎程序的潜在重新激活 由DSS治疗诱导。我们将测试胚胎或损伤相关间充质刺激的能力 可塑性，定义为成年人在更多功能状态的授权祖细胞的脱不同 上皮。第三，我们将采用一种合成方法来重建结肠周围图案的间充质信号 球体，我们将使用这种新模型来测试两极化Wnt信号的安全性来驱动结肠加密 体外格式。总体而言，我们的项目利用体内研究，体外模型，合成的组合 工程和生物信息学方法。通过高度使这个全面的范围成为可能 UCSF的协作环境以及两名主要研究人员的产品合作伙伴关系 这个项目。该提案的发现将大大提高我们对结肠发展的基本知识 和再生，突出了间质上皮串扰的动力学。这将有意义 除了我们对结肠生物学的基本理解之外，还将为发现治疗铺平道路 治疗和调节结肠疾病的策略。