Intestinal Stem Cell Survival and Renewal Coordinately Regulated by PUMA and p21

PUMA 和 p21 协调调节肠道干细胞的存活和更新

• 批准号: 8700642
• 负责人: Jian Yu
• 金额: $ 7.93万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8700642
• 关键词: Abdomen; Adult; Animals; Apoptosis; Biological Assay; Bone Marrow; Cell Cycle Arrest; Cell Lineage; Cell Separation; Cell Survival; Cells; Columnar Cell; Complication; Cultured Cells; DNA Damage; Data; Dose; Endothelial Cells; Genetic; Hematopoietic; Hematopoietic System; Histopathology; Home environment; Homeostasis; In Vitro; Injury; Intestines; Knock-out; Label; LacZ Genes; Lead; Mediating; Mediator of activation protein; Modeling; Molecular; Mus; Myofibroblast; Natural regeneration; Organ; Orphan; Paneth Cells; Pelvis; Play; Polycomb; Proteins; Radiation; Radiation Tolerance; Radiation induced damage; Radiation therapy; Reporter; Role; Small Intestines; Stem cells; System; The Sun; Therapeutic; Time; Tissues; Transplantation; Work; base; cell type; combat; defined contribution; effective therapy; intestinal epithelium; irradiation; mouse model; novel; oncoprotein p21; programs; public health relevance; receptor; reconstitution; repaired; response; stem cell biology

DESCRIPTION (provided by applicant): Adult tissue stem cells are believed to be responsible for maintaining tissue homeostasis and regeneration following injury in most tissues. The small intestine is one of the most sensitive organs to radiation-induced damage, which is the major complication in abdominal and pelvic radiotherapy with no effective treatment. The intestinal stem cells (ISCs) are located at or near the bottom of crypts in a unique yet poorly defined microenvironment ("niche") composed of several cell types derived from the bone marrow (BM). However, their identity remained elusive until recently. Genetic evidence demonstrated that the crypt-based columnar cells (CBCs) interspersed among Paneth cells marked by Lgr5 and some +4 cells immediately above Paneth cells marked by Bmi-1, represent perhaps distinct subsets of ISCs. Therefore, how their functions are regulated by intrinsic and extrinsic (niche-related) programs to participate in intestinal regeneration following injury is poorly understood. Work from us and others indicate that the BH3-only protein PUMA and cyclin-dependent kinase inhibitor p21 regulate the survival and regeneration of intestinal and hematopoietic systems, and the bone marrow modulates intestinal radiosensitivity. We hypothesize that both apoptosis and cell cycle arrest critically regulate the survival and regeneration of intestinal stem cells following radiation through intrinsic and extrinsic mechanisms. In this proposal, we will combine a novel ISC lineage marking and tracing mouse model with various knockout and transplantation models to 1) define the role of ISCs in crypt regeneration following radiation and develop assays for their isolation and characterization; 2) demonstrate a potential coordination of PUMA and p21 in modulating the survival and regeneration of ISCs; and 3) define the potential bone marrow contributions to intestinal regeneration. We believe that our studies can lead to a better understanding of ISC biology, and new models and strategies for isolation, characterization and manipulation of these critical cells for therapeutic purposes. PUBLIC HEALTH RELEVANCE: The proposed studies aim to determine the underlying molecular mechanisms governing the injury, survival and regeneration of intestinal stem cells following radiation, and to develop assays for their isolation and characterization. Such studies can help devise strategies to combat intestinal damage caused by radiotherapy or accidental radiation exposure.

描述（由申请人提供）：据信成年组织干细胞负责在大多数组织中损伤后维持组织稳态和再生。小肠是对辐射诱导的损伤最敏感的器官之一，这是腹部和骨盆放射疗法的主要并发症，没有有效的治疗。肠道干细胞（ISC）位于隐窝的底部或附近，在独特但定义较差的微环境（“利基”）中，该环境由骨髓（BM）衍生的几种细胞类型组成。但是，直到最近，他们的身份仍然难以捉摸。遗传证据表明，基于隐窝的柱状细胞（CBC）插入了以LGR5和BMI-1标记的Paneth细胞上方标记的Paneth细胞中散布的，代表了ISC的不同子集。因此，对损伤后的固有和外在和外在（小众相关）程序的调节如何调节其功能。我们和其他人的工作表明，仅BH3蛋白PUMA和细胞周期蛋白依赖性激酶抑制剂p21调节肠道和造血系统的存活和再生，骨髓调节肠道放射性启发性。我们假设凋亡和细胞周期停滞批判性地调节了通过固有和外在机制辐射后肠道干细胞的存活和再生。在此提案中，我们将将新颖的ISC谱系标记和追踪小鼠模型与各种敲除和移植模型相结合至1）定义ISC在辐射后的隐窝再生中的作用，并为其隔离和表征开发测定； 2）在调节ISC的生存和再生中，证明了PUMA和P21的潜在协调； 3）定义潜在的骨髓对肠再生的贡献。我们认为，我们的研究可以更好地理解ISC生物学，以及用于治疗目的的这些关键细胞的隔离，表征和操纵的新模型和策略。 公共卫生相关性：拟议的研究旨在确定辐射后肠道干细胞的损伤，生存和再生的基本分子机制，并开发用于隔离和表征的测定法。这样的研究可以帮助制定策略，以应对放射疗法或意外辐射暴露引起的肠道损害。