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

描述（由申请人提供）：以持续更新为特征，肠道为成体干细胞的研究提供了一个优雅的系统-这些细胞可以自我更新和再生，因此在再生医学方面有很大的希望。研究提供了重要的见解，关于能力的肠干细胞（ISCs）的自我修复或愈合 以及持续的维持和分化成各种肠细胞类型。然而，需要大量的研究来更好地了解ISCs亚群的不同状态和功能以及它们是如何被调节的。在其他组织中，如头发和血液，静止的干细胞亚群是至关重要的，特别是对于长期维持干细胞库和再生响应压力或损伤。然而，在肠道中，真正的静止ISCs是否存在和/或它们驻留在哪里是一个有争议的问题。这项研究提出，真正的静止ISCs与活跃ISCs共存。活性亚群负责肠上皮细胞谱系的产生和再生，而静止亚群作为“储备”库来补充失去的活性ISCs和受损组织。本研究的目的是进一步研究+4静止ISC，特别是，并确定潜在的分子调控的静止和活跃的ISC亚群。所使用的方法将包括：表面标记和体外培养以及体内谱系追踪测定，以在稳定和应激条件下鉴定和表征静止的ISCs; RNA测序，以确定某些因素如何以及在何处产生。（卷曲5和/或7）介导信号传导（非规范和/或规范Wnt），其控制ISC维持、激活、自我更新和定位;和遗传小鼠模型，以研究蛋白质失活对静止和活动ISCs的影响。了解静止和活动ISCs的状态和命运的信号调节可以发现治疗肠道疾病的潜在治疗靶点。 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ISC驱动的组织再生至关重要的肠道疾病以及筛选靶向癌症干细胞的药物开辟道路。识别、分离和表征ISCs的能力对于治疗进展（特别是组织替代）以及增强对肠道疾病的发展、预防和治愈的理解至关重要。

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.