Roles of BCL11A in γ-globin repression

项目总结 在成人红系细胞中，胎儿γ-珠蛋白是如何关闭的，这是珠蛋白转换的核心问题。一个更清晰的 对潜在机制的了解构成了临床重新激活胎儿血红蛋白(HBF)的基础。 一种治疗β-血红蛋白疾病的方法，如镰状细胞病和β-地中海贫血，其中两种 世界上最流行的遗传病。以前的研究已经确立了BCL11A作为主要的 γ-珠蛋白的抑制物，但其机制仍不清楚。本研究以机械解剖为主要研究对象。 BCL11A如何关闭γ-珠蛋白的表达，以及参与珠蛋白开关的新因素的鉴定。 我的广泛和长期目标是加强我们对发育基因控制的基本理解，以及 从而加快药物发现，造福公众健康。 在指导阶段，将利用强大的新功能研究BCL11A的详细作用机制 先进的技术。AIM 1问BCL11A如何与γ-珠蛋白转录激活子竞争 推动者。使用SMASH-BCL11A降解系统能够对BCL11A降解进行可逆控制， 与基因敲除相反，在基因敲除中，人们只能在很长一段时间后才能检查丢失的影响。 将使用Cut&Run高分辨率地定位BCL11A和转录激活子结合 控制BCL11A的降解和重新表达。AIM2研究了BCL11A在组织染色质中的作用 使用Capture-3C-Seq在β-珠蛋白基因座上的相互作用，它以高分辨率绘制染色质相互作用图 和敏感。目的3将探索相分离现象的功能作用的本质 BCL11A无序区(IDR)。相分离是最近的一项发现，它代表了一种范式 改变对生物分子如何在体内发挥其功能的看法。我的研究将集中在BCL11A期 分离调节珠蛋白基因的表达。这些研究将提供关于BCL11A如何 促进珠蛋白转换，这是BCL11A治疗靶向治疗疾病的基础。 我的长期职业目标是成为表观遗传学领域的一名富有成效和影响力的科学家。 发育过程中和血液疾病中的转录基因控制。为了实现这一目标，我有 为该奖项的指导阶段制定了详细的科学和职业发展计划，即 旨在提高我作为调查员和组长的技能。我邀请了两位老牌和一位 初级调查员在我的咨询委员会中服务，并指导我的培训。拟议的活动包括课程、 会议和实践，主要在波士顿儿童医院和周围机构举行， 包括哈佛医学院和达纳-法伯癌症研究所。提供了一流的培训环境 来自这些机构的宝贵批评和我的导师和咨询委员会的支持以及 从这个K99奖将极大地促进那些必要的专业技能的发展，使 成功过渡到独立调查员。

PROJECT SUMMARY How fetal γ-globin is turned off in adult erythroid cells is a central question of the globin switch. A clearer understanding of the underlying mechanisms forms the basis for clinical reactivation of fetal hemoglobin (HbF) - a therapeutic approach to treat β-hemoglobin disorders such as sickle cell disease and β-thalassemia, two of the most prevalent genetic diseases worldwide. Previous studies have established the role of BCL11A as a major repressor of γ-globin, but the mechanism remains elusive. This research focuses on mechanistic dissection of how BCL11A turns off γ-globin expression, and the identification of novel factors involved in the globin switch. My broad and long-term objectives are to enhance our basic understanding of developmental gene control, and thereby accelerate drug discovery to benefit public health. During the mentored phase, the detailed mechanism of BCL11A action will be studied using powerful newly developed technologies. Aim 1 asks how BCL11A competes with transcriptional activators at the γ-globin promoters. Use of the SMASh-BCL11A degradation system enables reversible control of BCL11A degradation, as opposed to gene knockouts where one can only examine effects of loss after a long intervening time period. CUT&RUN will be employed to map BCL11A and transcriptional activator binding at high resolution during controlled BCL11A degradation and re-expression. Aim2 examines the role of BCL11A in organizing chromatin interactions at the β-globin locus using CAPTURE-3C-seq, which maps chromatin interactions at high resolution and sensitivity. Aim 3 will explore the functional roles of the phase separation phenomenon of the Intrinsically Disordered Regions (IDRs) of BCL11A. Phase separation is a recent discovery that represents a paradigm changing view of how biomolecules carry out their function in vivo. My study will focus on whether BCL11A phase separation regulates globin gene expression. These studies will provide mechanistic insights of how BCL11A facilitates globin switching, which is the foundation of therapeutic targeting of BCL11A to treat diseases. My long-term career goal is to become a productive and impactful scientist in the field of epigenetic and transcriptional gene control during development and in hematological diseases. To achieve this goal, I have developed a detailed scientific and career development plan for the mentored phase of the award, which is designed to enhance my skills as both investigator and group leader. I have invited two established and one junior investigator to serve on my advisory committee and guide my training. Proposed activities include courses, meetings and practices, and mostly take place at Boston Children's Hospital and surrounding institutions, including Harvard Medical School and Dana-Farber Cancer Institute. The superb training environment provided by these institutions, the invaluable critique and support from my mentor and advisory committee, and the support from this K99 award will greatly advance development of those professional skills necessary to make a successful transition to an independent investigator.