Functional evaluation of clonal hematopoiesis of indeterminate potential (CHIP)

摘要 不确定潜能的克隆性造血是造血干细胞克隆的扩增 获得性突变被认为是由正常衰老或骨髓压力引起的。CHIP克隆人 患血癌、心血管疾病或缺血性中风的风险增加。我们的理解 CHIP进展的病理生理学研究仅限于大型下一代测序研究 在成年人中进行。因此，分子机制和克隆扩张和动态的背景下， CHIP突变，知之甚少。作为奥本实验室的博士后研究员， 博士Mitch韦斯，我将研究儿童患者CHIP扩张的病理生物学机制， 小鼠模型。在目标1中，我将使用靶向基因组测序来确定 在一个大型的，注释良好的儿科患者队列中研究CHIP的并发症。在这个目标下，我将帮助定义CHIP 研究不足的队列中的发病率，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 （HSC）表达不同的、共同的CHIP突变。我将确定不同的突变如何影响克隆 在不同的HSC区室扩张，以及慢性炎症和衰老如何影响HSC的行为， 不同的CHIP克隆在这一目标中，我将帮助确定携带CHIP相关突变的HSC克隆如何扩增， in vivo.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， 干细胞生物学和功能基因组学的新培训。圣裘德的技术和智力资源 儿童研究医院为我的训练提供了最佳的环境。除了特殊的 培训和进入国家的最先进的核心设施提供在圣裘德，我也将利用 在我的奖学金期间，我以机构和外部研讨会的形式获得了许多职业发展资源， 建立联系的机会、教学和指导的机会以及研讨会。我就能得到一个大的， 注释良好的临床队列、表征良好的动物模型和最先进的基因组测序， 流式细胞仪设备。我将与我们在计算生物学和生物统计学方面的合作者密切合作， 学习如何分析我的实验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，这将有助于我成为一名独立调查员。 我的研究将有助于定义允许CHIP克隆在体内持续和扩展的分子特征。更好的 了解CHIP克隆的病理生物学将有助于指导如何监测CHIP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.