Functional evaluation of clonal hematopoiesis of indeterminate potential (CHIP)

摘要 未确定潜能克隆性造血(CHIP)是造血干细胞(HSC)克隆的扩增 获得性突变被认为是由正常衰老或骨髓压力引起的。拥有克隆芯片的人 患血癌、心血管疾病或缺血性中风的风险增加。我们的理解 芯片进展的病理生理学研究仅限于大型下一代测序研究 在成年人身上表演。因此，分子机制和克隆扩张和动力学背景下 对于芯片突变，人们知之甚少。作为奥本格实验室的博士后研究员，由 Mitch Weiss博士，我将研究儿科患者芯片扩展的病理生物学机制，并使用 老鼠模型。在目标1中，我将使用定向基因组测序来定义发病率和临床 CHIP在一大群有良好注解的儿科患者中的并发症。在这个目标中，我将帮助定义芯片 在研究较少的队列中的发病率，并扩展我在计算生物学和功能基因组学方面的培训。 在目标2中，我将使用分子条形码来标记和跟踪造血干细胞的克隆动力学 (HSCs)表达不同的、常见的芯片突变。我将确定不同的突变如何影响克隆 慢性炎症和衰老如何影响HSC的行为 不同的芯片克隆。在这个目标中，我将帮助定义含有芯片相关突变的HSC克隆是如何扩展的 在活体内。总的来说，这些目标借鉴了我以前的分子和计算生物学经验，并提供了 干细胞生物学和功能基因组学方面的新培训。圣犹大的技术和智力资源 儿童研究医院为我的训练提供了最佳的环境。除了特殊的 培训和使用圣裘德最先进的核心设施，我还将利用 在我担任研究员期间，以机构和外部研讨会的形式提供了许多职业发展资源， 网络机会、教学和指导机会以及研讨会。我将有机会接触到一个大型的， 注解良好的临床队列，特征良好的动物模型，以及最先进的基因组测序和 流式细胞仪设备。我将与我们在计算生物学和生物统计学领域的合作者密切合作，以 学习如何分析我的实验生成的数据。我将与经验丰富、敬业的导师一起工作 并将获得职业发展机会，这将帮助我成为一名独立调查员。 我的研究将有助于确定允许芯片克隆在体内持续和扩展的分子特征。更好的 了解芯片克隆的病理生物学将有助于指导如何监测和通知芯片进展 所有年龄段患者的治疗考虑。

ABSTRACT Clonal hematopoiesis of indeterminate potential (CHIP) is the expansion of hematopoietic stem cell (HSC) clones with acquired mutations thought to be caused by normal aging or bone marrow stress. People with CHIP clones are at increased risk of developing a blood cancer, cardiovascular disease or ischemic stroke. Our understanding of the pathophysiology of CHIP progression has been limited to large next generation sequencing studies performed in adults. Therefore, the molecular mechanisms and clonal expansion and dynamics in the context of CHIP mutations, are poorly understood. As a postdoctoral fellow in the Obeng laboratory and co-sponsored by Dr. Mitch Weiss, I will study the pathobiological mechanisms of CHIP expansion in pediatric patients and using mouse models. In Aim 1, I will use targeted genomic sequencing to define the incidence and clinical complications of CHIP in a large, well-annotated cohort of pediatric patients. In this aim, I will help define CHIP incidence in poorly studied cohort and expand on my training in computational biology and functional genomics. In Aim 2, I will use molecular barcoding to label and track the clonal dynamics of hematopoietic stem cells (HSCs) expressing different, common CHIP mutations. I will determine how different mutations affect clonal expansion in different HSC compartments and how chronic inflammation and aging affect the behavior of different CHIP clones. In this aim, I will help define how HSC clones harboring CHIP-related mutations expand in vivo. Collectively, these aims draw upon my prior molecular and computational biology experience and provide new training in stem cell biology and functional genomics. The technical and intellectual resources at St. Jude Children's Research Hospital provide the optimal environment for my training. In addition to the exceptional training and access to the state-of-the-art core facilities available at St. Jude, I will also take advantage of numerous career development resources during my fellowship in the form of institutional and outside workshops, networking opportunities, teaching and mentoring opportunities, and seminars. I will have access to a large, well-annotated clinical cohort, well-characterized animal models, and state-of-the-art genome sequencing and flow cytometry facilities. I will work closely with our collaborators in Computational Biology and Biostatistics to learn how to analyze the data generated by my experiments. I will work with experienced, dedicated mentors and will have access to career development opportunities that will help me become an independent investigator. My studies will help define the molecular features that allow CHIP clones to persist and expand in vivo. A better understanding of the pathobiology of CHIP clones will help guide how CHIP progression is monitored and inform therapeutic considerations in patients of all ages.