Characterization of Age-related Changes in Stem Cell Behavior

干细胞行为与年龄相关的变化的表征

• 批准号: 8046332
• 负责人: DANA LEANNE JONES
• 金额: $ 36.61万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8046332
• 关键词: Address; Adult; Age; Aging; Aging-Related Process; Antibodies; Biological Assay; Blood; Bromodeoxyuridine; Cell Adhesion; Cell Maintenance; Cell Proliferation; Cell division; Cell physiology; Cells; Characteristics; Disease; Drosophila genus; Environment; Exhibits; Failure; Female; Gene Expression; Gene Proteins; Genetic; Germ Cells; Germ Lines; Goals; Gonadal structure; Histone H3; Homeostasis; Immunofluorescence Immunologic; Immunofluorescence Microscopy; In Situ; Individual; Intrinsic factor; Life; Longevity; Maintenance; Measures; Methods; Mitotic; Mitotic Recombination; Molecular; Muscle; Organ; Pathway interactions; Phase-Contrast Microscopy; Regenerative Medicine; Replacement Therapy; Research; Research Personnel; Role; Signal Transduction; Skin; Spermatogenesis; Spermatogonia; Staging; Stem cells; Supporting Cell; System; Testis; Time; Tissues; To specify; age related; cell behavior; cell type; daughter cell; fly; gain of function; in vivo; insight; loss of function; male; mutant; older patient; organ regeneration; programs; protein expression; regenerative; self-renewal; sperm cell; stem; stem cell division; stem cell niche; technique development

DESCRIPTION (provided by applicant): Stem cells provide for the maintenance and regeneration of organs and tissues throughout life, and loss of the regenerative capacity of tissues such as blood and muscle has been attributed to decreased stem cell activity. The ability of stem cells to contribute to tissue homeostasis depends on the unique ability to generate both new stem cells (self-renewal) as well as specialized cell types (differentiation). The Aims of this proposal are to analyze changes in stem cell behavior during the aging process and to identify molecular mechanisms regulating these changes. We will also investigate the relationship between pathways regulating longevity and age-related changes to stem cells and the stem cell niche. Methods: We previously characterized the germ line stem cell (GSC) niche in the Drosophila testis, providing a system to study the intrinsic and extrinsic factors controlling stem cell behavior in vivo. Testes from males maintained with females will be analyzed for changes in stem cell behavior at 1, 30, and 50 days post-eclosion. Using readily available markers, we will analyze changes in gene expression and protein expression and localization in stem cells and niche cells using in situ analysis and immunofluorescence. GSC division will be assayed using BrdU incorporation as well as by marking cells through mitotic recombination. GSC behavior will be analyzed in wild type flies, flies in which the JAK-STAT pathway, which is necessary and sufficient for stem cell self-renewal, has been modulated, and in flies that have been characterized as being long-lived. Conclusions: In addition to providing insights into tissue homeostasis, studies focusing on age-related changes in stem cells and their specialized microenvironments, known as stem cell niches, will help to identify and overcome unique hurdles in the manipulation of tissue stem cells derived from older patients and to facilitate the development of techniques for regenerative medicine to treat aging-related diseases. Lay Summary: Evidence suggests that stem cell function decreases during the aging process, and the primary consequence is loss of tissue function. Tissue replacement therapies will rely heavily on expanding tissue stem cells in culture. This project seeks to characterize age-related changes in stem cells and the support cells that nurture them to facilitate the expansion and maintenance of stem cells in culture, an essential first step in the utilization of stem cells to treat aging-related diseases.

描述（由申请人提供）：干细胞在整个生命过程中提供器官和组织的维持和再生，血液和肌肉等组织再生能力的丧失归因于干细胞活性的降低。干细胞促进组织稳态的能力取决于产生新干细胞（自我更新）和特化细胞类型（分化）的独特能力。该提案的目的是分析衰老过程中干细胞行为的变化，并确定调节这些变化的分子机制。我们还将研究干细胞和干细胞生态位的长寿调节途径和年龄相关变化之间的关系。方法：我们之前对果蝇睾丸中的生殖系干细胞（GSC）生态位进行了表征，提供了一个系统来研究控制体内干细胞行为的内在和外在因素。与雌性一起饲养的雄性睾丸将在羽化后 1、30 和 50 天分析干细胞行为的变化。使用现成的标记，我们将使用原位分析和免疫荧光分析干细胞和利基细胞中基因表达和蛋白质表达以及定位的变化。 GSC 分裂将使用 BrdU 掺入以及通过有丝分裂重组标记细胞来测定。我们将在野生型果蝇、JAK-STAT 途径（干细胞自我更新所必需且充分）已被调节的果蝇以及已被定性为长寿的果蝇中分析 GSC 行为。结论：除了提供对组织稳态的见解外，关注干细胞及其特殊微环境（称为干细胞生态位）与年龄相关的变化的研究将有助于识别和克服操纵老年患者组织干细胞的独特障碍，并促进治疗衰老相关疾病的再生医学技术的发展。摘要：有证据表明，干细胞功能在衰老过程中下降，主要后果是组织功能丧失。组织替代疗法将在很大程度上依赖于培养中组织干细胞的扩增。该项目旨在描述干细胞和培育干细胞的支持细胞与年龄相关的变化，以促进干细胞在培养物中的扩增和维持，这是利用干细胞治疗衰老相关疾病的重要第一步。