Photoresponsive materials to study matricellular signaling dynamics during crypt formation and fission

用于研究隐窝形成和裂变过程中基质细胞信号动力学的光响应材料

基本信息

批准号：
10737202
负责人：
KRISTI S. ANSETH
金额：
$ 57.49万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2028-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10737202
关键词：
Adhesives Alleles Biomechanics Cell Compartmentation Cell Shape Cell physiology Cell secretion Cells Cellularity Complex Crohn&apos s disease Daughter Development Digestion Disease Engineering Epithelial Cells Epithelium Event Extracellular Matrix Geometry Health Homeostasis Hydrogels In Vitro Infection Inflammatory Bowel Diseases Injury Integrin Binding Intestinal Diseases Intestines LGR5 gene Label Letters Light Location Maintenance Malignant Neoplasms Mediating Membrane Microscopy Modeling Modulus Mus Myosin Heavy Chains Natural regeneration Nuclear Nutrient Organoids Paneth Cells Parents Pattern Peptides Positioning Attribute Process Property Protein Secretion Reporter Reproducibility Research Resolution Role Shapes Signal Transduction Small Intestines Structure Tamoxifen Testing Three-Dimensional Imaging Time Tissues Ulcerative Colitis Validation Villus base cell type crypt cell density experimental study extracellular in vitro Model in vivo in vivo Model injury and repair innovation intercellular communication intestinal crypt knock-down mechanotransduction mouse model nutrient absorption repaired response spatiotemporal stem cell niche stem cells tissue regeneration two-photon unnatural amino acids

项目摘要

ABSTRACT The small intestine is lined with a single layer of epithelial cells that is organized into crypt-villus units. These epithelial cells perform multiple functions, such as aiding in digestion, nutrient absorbance, and serving as a barrier, but these processes also damage the epithelial layer. As a result, intestinal stem cell (ISCs) located near the base of crypts are responsible for the constant renewal and rapid replenishment of all intestinal cell types, but the crypts themselves also require maintenance through a process called fission. Crypt fission occurs when a single intestinal crypt divides and generates a new daughter crypt, which is essential for maintaining overall crypt density and intestinal health. Despite the important role of crypt fission in injury-induced crypt regeneration and its mis-regulation in intestinal diseases (e.g., inflammatory bowel disease), little is known about the mechanisms of how crypt fission is initiated, progresses through crypt bifurcation, and is regulated under different pathophysiological conditions. Thus, the proposed research aims to use photoresponsive hydrogels for the culture of intestinal organoids and then develop a robust and predictable in vitro model of crypt fission events. Our innovative materials and reductionist approach will allow us to precisely tune the properties of the ISC niche and understand how epithelial cells and ECM signaling contribute to crypt formation and fission. We will test hypotheses related to the role of crypt cells and matricellular signaling and its effects on organoid symmetry breaking, crypt formation, crypt fission, and the potential compensatory cellular response to local tissue damage. Specifically, we aim to: 1. Investigate the role of spatiotemporally varying cell-matrix on real time crypt formation in intestinal organoids. 2. Iteratively pattern intestinal organoid-laden hydrogels to study the role of the ECM interactions in guiding crypt fission. and 3. Spatiotemporally regulate crypt cell composition to investigate its role in crypt fission.

抽象的小肠内衬有单层上皮细胞，这些上皮细胞被组织成隐窝绒毛单位。这些上皮细胞执行多种功能，例如帮助消化、营养吸收和充当屏障，但这些细胞过程也会损伤上皮层。因此，位于隐窝底部附近的肠干细胞（ISC）负责所有肠道细胞类型的不断更新和快速补充，但隐窝本身也负责需要通过称为裂变的过程进行维护。当单个肠隐窝分裂时，就会发生隐窝裂变产生一个新的子隐窝，这对于维持整体隐窝密度和肠道健康至关重要。尽管隐窝裂变在损伤诱导的隐窝再生中的重要作用及其在肠道疾病中的错误调节（例如，炎症性肠病），关于隐窝裂变如何启动、进展的机制知之甚少隐窝分叉，并在不同的病理生理条件下受到调节。因此，拟议的研究旨在使用用于培养肠道类器官的光响应水凝胶，然后开发稳健且可预测的体外模型地穴裂变事件。我们的创新材料和还原论方法将使我们能够精确调整 ISC 生态位并了解上皮细胞和 ECM 信号如何促进隐窝形成和裂变。我们将测试与隐窝细胞和基质细胞信号传导的作用及其对类器官对称性破坏的影响相关的假设，隐窝形成、隐窝裂变以及对局部组织损伤的潜在代偿性细胞反应。具体来说，我们目的： 1. 研究时空变化的细胞基质对肠道类器官实时隐窝形成的作用。 2. 迭代地模拟充满肠道类器官的水凝胶，以研究 ECM 相互作用在引导隐窝裂变中的作用。 3.时空调节隐窝细胞组成以研究其在隐窝裂变中的作用。