Analysis of Stem Cell Dynamics and Differentiation By Cellular Barcoding

描述（由申请人提供） 摘要：发达国家不断变化的人口统计数据强调了再生医学方法对抗退行性疾病的临床和经济负担的必要性。因此，对组织如何由其常驻干细胞正常维持的基本理解是追求再生方法的关键。虽然在过去的二十年里，通过使用传统的实验方法已经获得了大量的知识，但这些技术的局限性和缺点使我们无法完全了解干细胞的功能，特别是在体内环境中。我的新创新者计划的目标是为干细胞生物学研究开发一种新的实验范式。在该模型中，群体中的个体干细胞在原位被独特地和遗传地标记，并且这些独特的遗传标记或条形码然后可以用于动态地监测个体干细胞活性、寿命和高度复杂群体中随时间的分化。我们相信，这个模型可以为我们提供一个复杂而动态的干细胞系统内部运作的前所未有的高分辨率图像，并使我们能够回答长期存在的生物学问题。我们的研究结果最终可能揭示在老年或其他疾病背景下受到干扰的干细胞维持的保守机制。虽然这项提议将解决正常和恶性造血（造血）干细胞室的生物学问题，但我们模型的模块化性质使其易于适应任何组织。我们的模型也适用于衰老和免疫学问题的研究。 公共卫生相关性：该提案将使我们能够了解造血干细胞在生物体中的功能。从这些研究中获得的知识将使我们能够了解干细胞功能如何在骨髓衰竭综合征，衰老和血液癌症等情况下受到影响。

DESCRIPTION (Provided by the applicant) Abstract: The changing demographics of developed nations underscores the need for regenerative medicine approaches to combat the clinical and financial burden of degenerative diseases. The basic understanding of how tissues are normally maintained by their resident stem cells is, therefore, key for pursuing regenerative approaches. Though a great deal of knowledge has been gained through the use of traditional experimental approaches over the past two decades, limitations and drawbacks of these techniques have precluded us from gaining a complete understanding of stem cell function, particularly in the in vivo setting. The goal of my New Innovator proposal is to develop a novel experimental paradigm for the study of stem cell biology. In this model, individual stem cells in a population are uniquely and genetically tagged in situ and these unique genetic tags, or barcodes, can then be used to dynamically monitor individual stem cell activity, lifespan, and differentiation in highly complex populations over time. We believe that this model can give us an unprecedented highresolution picture of the inner workings of a complex and dynamic stem cell system, and allow us to answer long-standing biological questions. Our findings ultimately may uncover conserved mechanisms of stem cell maintenance that are perturbed in old age or other disease contexts. Though this proposal will address the biology of the normal and malignant hematopietic (bloodforming) stem cell compartments, the modular nature of our model makes easily adaptable to any tissue. Our model is also suitable, among other things, for the study of aging and immunological problems. Public Health Relevance: This proposal will allow us to understand how blood-forming stem cells function in an organism. The knowledge gained from these studies will allow us to understand how stem cell function is affected in conditions such as bone marrow failure syndromes, aging, and cancers of the blood.