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结构突显了再生医学方法的必要性，以对抗退行性疾病的临床和经济负担。因此，对组织如何正常地由其常驻干细胞维持的基本了解，是寻求再生方法的关键。尽管在过去的二十年里，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陷阻碍了我们对干细胞功能的完全了解，特别是在活体环境中。我的新创新者提案的目标是为干细胞生物学研究开发一种新的实验范式。在这个模型中，种群中的单个干细胞被唯一地和原位地遗传标记，然后这些独特的遗传标签或条形码可以被用来动态监测个体干细胞的活动、寿命和随着时间的推移在高度复杂的种群中的分化。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工作原理的前所未有的高分辨率图像，并允许我们回答长期存在的生物学问题。我们的发现最终可能揭示干细胞维持的保守机制，这些机制在老年或其他疾病背景下受到干扰。虽然这项建议将解决正常和恶性造血(造血)干细胞隔间的生物学问题，但我们模型的模块化性质使其很容易适应任何组织。我们的模型也适用于衰老和免疫学问题的研究。 与公共卫生相关：这项提议将使我们能够了解造血干细胞如何在生物体中发挥作用。从这些研究中获得的知识将使我们了解在骨髓衰竭综合征、衰老和血癌等情况下干细胞功能是如何受到影响的。

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.