Cellular, Molecular, and Functional Characterization of Quiescent/Active Intestin

静态/活跃肠的细胞、分子和功能表征

• 批准号: 8774720
• 负责人: LINHENG LI
• 金额: $ 36.45万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 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 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8774720
• 关键词: Accounting; Affect; American; Biological Assay; Blood; CD44 gene; Candidate Disease Gene; Cell Maintenance; Cells; Colitis; Columnar Cell; Control Groups; Crohn' s disease; Data; Development; Digestive System Disorders; Drug resistance; Endocrine; Epithelial; Funding; GRP78 gene; Gene Expression; Generations; Genes; Genetic; Goals; Hair; Healed; Homeostasis; Hypoxia; In Vitro; Injury; Intestinal Cancer; Intestinal Diseases; Intestines; Knowledge; Label; Ligands; Location; Maintenance; Malignant Neoplasms; Mediating; Methods; Molecular; Molecular Profiling; Natural regeneration; Normal tissue morphology; Outcome; Paneth Cells; Pathway interactions; Patients; Play; Population; Positioning Attribute; Preclinical Drug Evaluation; Prevention; Proteins; Public Health; RNA Sequences; Regenerative Medicine; Regulation; Relapse; Research; Resistance; Role; Signal Pathway; Signal Transduction; Sorting - Cell Movement; Stem cells; Stress; Surface; System; Therapeutic; Tissues; Treatment Efficacy; Wound Healing; adult stem cell; base; cancer stem cell; cell behavior; cell injury; cell type; combinatorial; cost; crypt cell; healing; improved; in vivo; insight; mouse model; progenitor; public health relevance; receptor; repaired; response; self-renewal; stem cell biology; stem cell population; therapeutic target; tissue regeneration; transcriptome sequencing

DESCRIPTION (provided by applicant): Characterized by continual turnover, the intestine provides an elegant system for the study of adult stem cells - - cells which self-renew and regenerate and, thus, hold great promise for regenerative medicine. Studies have provided important insight regarding the capacity of intestinal stem cells (ISCs) for self-repair or healing as well as ongoing maintenance and differentiation into all the various intestinal cell types. However, considerable research is needed to better understand the different states and functions of subpopulations of ISCs and how these are regulated. In other tissues, such as hair and blood, the quiescent stem cell subpopulation is known to be critical, especially for long-term maintenance of the stem cell pool and for regeneration in response to stress or injury. However, in intestine, it is a matter of debate whether bona fide quiescent ISCs exist and/or where they reside. This research proposes that bona fide quiescent ISCs coexist with active ISCs. The active subpopulation accounts for generation and regeneration of intestinal epithelial lineages, and the quiescent subpopulation functions as a 'reserve' pool to replenish lost active ISCs and damaged tissues. The goal of this research is to further investigate +4 quiescent ISCs, in particular, and to determine potential molecular regulation of quiescent and active ISC subpopulations. Methods to be used will include: surface markers and in vitro culture and in vivo lineage tracing assays to identify and characterize quiescent ISCs under stable and stressed conditions; RNA sequencing to determine how and where certain factors (Frizzled5 and/or 7) mediate signaling (noncanonical and/or canonical Wnt) that govern ISC maintenance, activation, self- renewal and location; and genetic mouse models to study the effect of protein inactivation on quiescent and active ISCs. Understanding signaling regulation of the state and fate of quiescent and active ISCs can uncover potential therapeutic targets for treating intestinal disorders, including cancer. If this goal can be achieved, it will open avenues not only for advancing the study of ISC behavior, but also for treating intestinal disorders in which ISC-driven tissue regeneration is essential and for screening drugs to target cancer stem cells. The ability to identify and isolate and characterize ISCs is critical for therapeutic advancements, especially tissue replacement, and for enhanced understanding of the development, prevention, and cure of intestinal disease.

描述（由申请人提供）：肠道以不断更新为特征，为研究成体干细胞提供了一个优雅的系统，这些细胞具有自我更新和再生能力，因此在再生医学方面具有广阔的前景。研究提供了有关肠道干细胞 (ISC) 自我修复或愈合能力的重要见解 以及持续的维持和分化为所有各种肠道细胞类型。然而，需要大量研究来更好地了解 ISC 亚群的不同状态和功能以及它们是如何受到监管的。在头发和血液等其他组织中，已知静止干细胞亚群至关重要，特别是对于干细胞库的长期维持以及响应压力或损伤的再生。然而，在肠道中，真正的静态 ISC 是否存在和/或它们驻留在何处仍存在争议。这项研究提出，真正的静态 ISC 与活跃 ISC 共存。活跃的亚群负责肠上皮谱系的生成和再生，而静止的亚群则充当“储备”池，以补充丢失的活性 ISC 和受损组织。本研究的目的是进一步研究 +4 静态 ISC，特别是确定静态和活性 ISC 亚群的潜在分子调节。使用的方法包括：表面标记以及体外培养和体内谱系追踪测定，以识别和表征稳定和应激条件下的静态 ISC； RNA测序以确定某些因子（Frizzled5和/或7）如何以及在何处介导控制ISC维持、激活、自我更新和定位的信号传导（非规范和/或规范Wnt）；和遗传小鼠模型来研究蛋白质失活对静止和活跃 ISC 的影响。了解静止和活跃 ISC 的状态和命运的信号调节可以发现治疗肠道的潜在治疗靶点 疾病，包括癌症。如果这一目标能够实现，它不仅将为推进 ISC 行为的研究开辟道路，而且还将为治疗肠道疾病（其中 ISC 驱动的组织再生至关重要）以及筛选针对癌症干细胞的药物开辟道路。识别、分离和表征 ISC 的能力对于治疗进展（尤其是组织替代）以及增强对肠道疾病的发展、预防和治疗的了解至关重要。