Age Compromises Novel Motility and Repair Functions in Stem Cell Niche of Intestinal Crypts

年龄会损害肠隐窝干细胞生态位的新活力和修复功能

• 批准号: 9753843
• 负责人: Nozomi Nishimura
• 金额: $ 20.61万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9753843
• 关键词: Ablation; Address; Age; Aging; Animals; Antibodies; Autopsy; Biological Assay; Blood Vessels; Bromodeoxyuridine; Cell Count; Cell Nucleus; Cell Proliferation; Cell division; Cell physiology; Cells; Chronic; Colitis; Complement; Coupled; Development; Disease; Epithelial Cells; Epithelium; Equilibrium; Excision; Goals; Grant; Green Fluorescent Proteins; Health; Homeostasis; Image; Imagery; Immunofluorescence Immunologic; Impairment; Implant; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Injury; Intervention; Intestines; LGR5 gene; Label; Lasers; Left; Link; Location; Longevity; Measurement; Mechanics; Methods; Modeling; Molecular; Monitor; Motion; Mucous Membrane; Mus; Natural regeneration; Optics; Organism; Pathologic; Pathology; Pathway interactions; Pattern; Penetration; Positioning Attribute; Preparation; Process; Proteins; Reactive Oxygen Species; Recovery; Shapes; Sirolimus; Small Intestines; Stains; Stem cells; Techniques; Testing; Time; Tissues; Variant; age effect; base; cell behavior; cell injury; cell motility; cell type; crypt cell; daughter cell; expectation; imaging approach; imaging modality; improved; in vivo; in vivo imaging; injured; intestinal crypt; intestinal epithelium; macrophage; microscopic imaging; multiphoton microscopy; next generation; novel; preservation; repaired; response; response to injury; seal; self-renewal; stem cell niche; stem cell population; tissue repair; tool; tumorigenesis; young adult

As the primary controllers of epithelium regeneration, intestinal stem cells (ISCs) at the bottom of the crypt must maintain a balance between self-renewal and differentiation. However, it is still unclear how the stem cells maintain tissue homeostasis in response to daily variations in cell numbers or after injury. It is also well known that the ability to repair damage is reduced in aging, but it is not known what mechanism(s) underly this process. We have recently developed a chronic preparation for in vivo imaging with multiphoton microscopy which allows the monitoring the ISC niche in real-time in mice expressing green fluorescent protein in Lgr5+ ISCs. The goal is to directly track and identify how stem cells maintain their balance in the intestinal crypts. Next generation multiphoton microscopy with an in vivo imaging preparation with femtosecond laser ablation is used to ablate individual cells of a specific type to perturb the crypt. Time lapse imaging captures changes in cell number, position, motion and marker expression to identify how the various populations of stem cell respond. Upon ablation, the targeted cells lost their shape and moved out of the plane of the crypt base towards the intestinal lumen. Immediately adjacent cells appeared to move into the space left by the ablated cell, suggesting that the niche cells actively move around in response to the pattern disruption. This proposal tests the hypothesis that age and underlying pathology reduces the efficacy of these newly discovered dynamics, which can be rescued by age-delaying agents. The expectation is that these motions are involved in protecting the stem cells from damaging factors spilling from injured cells. The new optical tools have identified a potential new function of ISCs. In addition to generating daughter cells to replenish the epithelium, Lgr5+ ISCs appear to be mechanically active in eliminating damaged cells. This adds a new function to the repertoire of ISC actions. Collectively, the results suggest that there is an active process that involves cell migration in addition to cell division for maintaining homeostasis in the intestinal crypt and epithelium. !

作为上皮再生的主要控制器，肠道干细胞（ISC）处于隐窝底部 必须保持自我更新和差异之间的平衡。但是，目前尚不清楚茎如何 细胞响应细胞数量或损伤后的每日变化而保持组织稳态。也很好 知道修复损伤的能力在衰老中会降低，但尚不知道是什么机制 这个过程。我们最近开发了一种慢性准备，用于使用多光子进行体内成像 显微镜，可以在表达绿色荧光的小鼠中实时监测ISC利基市场 LGR5+ ISC中的蛋白质。目标是直接跟踪和确定干细胞如何保持其平衡 肠道隐窝。下一代多光子显微镜与体内成像制剂一起制备 飞秒激光消融用于消融特定类型的单个细胞以扰乱隐窝。时间流逝 成像捕获细胞数，位置，运动和标记表达式的变化，以识别各种 干细胞的种群反应。消融后，目标细胞失去了形状，并移出了飞机 朝向肠腔的隐窝底部。立即相邻的单元似乎移入空间 由烧蚀的细胞留下，表明利基细胞响应于模式而主动移动 破坏。该提案检验了以下假设：年龄和潜在的病理降低了这些的疗效 新发现的动力学，可以通过延迟的代理商来挽救。期望这些 动作涉及保护干细胞免受受伤细胞溢出的破坏因素的影响。新的 光学工具已经确定了ISC的潜在新功能。除了生成子细胞 补充上皮，LGR5+ ISC似乎在消除受损细胞方面具有机械活性。这 在ISC操作的曲目中添加了一个新功能。总体上，结果表明有一个活动 除细胞分裂外，还涉及细胞迁移以维持肠内稳态 地穴和上皮。 呢