Roles of BCL11A in γ-globin repression

BCL11A 在 γ-珠蛋白抑制中的作用

基本信息

批准号：
9892279
负责人：
Nan Liu
金额：
$ 9.15万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9892279
关键词：
Address Adult Advanced Development Advisory Committees Area Award Binding Boston CRISPR/Cas technology Cell Line ChIP-on-chip Chromatin Chromatin Structure Clinical Critiques Cultured Cells DNA Dana-Farber Cancer Institute Detection Development Development Plans Developmental Gene Disease Dissection Down-Regulation Drug Targeting Elements Enhancers Environment Epigenetic Process Erythroid Erythroid Cells Erythroid Progenitor Cells Fetal Hemoglobin Foundations Gene Cluster Gene Expression Gene Expression Profiling Gene Expression Regulation Genes Genetic Genetic Diseases Genetic Transcription Genome Globin Goals Hematological Disease Hemoglobin Hemoglobin concentration result Institution Intercistronic Region Investigation Knock-out Knockout Mice Laboratories Locus Control Region Maps Mentors Methods Modeling Mutation Pathway interactions Pediatric Hospitals Phase Play Proteins Proteomics Public Health Regulation Regulatory Element Repression Research Research Personnel Resolution Role Scientist Sickle Cell Anemia Site System Techniques Technology Therapeutic Time Tissue-Specific Gene Expression Training Transcription Coactivator Transcriptional Regulation Zinc Fingers beta Globin beta Thalassemia career career development clinical development design drug discovery fetal fetal reactivity gamma Globin genome wide association study hemoglobin B in vivo insight knockout gene medical schools meetings novel promoter skills small hairpin RNA therapeutic target

项目摘要

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.

项目概要胎儿γ-珠蛋白如何在成人红细胞中被关闭是珠蛋白开关的一个核心问题。了解潜在机制是胎儿血红蛋白 (HbF) 临床再激活的基础 - 一种治疗 β-血红蛋白疾病的治疗方法，例如镰状细胞病和 β-地中海贫血，这两种疾病先前的研究已确定 BCL11A 作为一种主要的遗传疾病。 γ-球蛋白的阻遏物，但其机制仍然难以捉摸。本研究重点是其机制剖析。 BCL11A 如何关闭 γ-珠蛋白表达，以及参与珠蛋白开关的新因子的鉴定。我的广泛而长期的目标是增强我们对发育基因控制的基本理解，并且从而加速药物发现，造福公众健康。在指导阶段，将使用强大的新方法来研究 BCL11A 作用的详细机制。目标 1 询问 BCL11A 如何与 γ-球蛋白上的转录激活剂竞争。使用 SMASh-BCL11A 降解系统可以对 BCL11A 降解进行可逆控制，与基因敲除相反，基因敲除只能在经过很长一段时间后才能检查损失的影响。 CUT&RUN 将用于在高分辨率过程中绘制 BCL11A 和转录激活剂结合图谱。受控的 BCL11A 降解和重新表达 Aim2 检查 BCL11A 在组织染色质中的作用。使用 CAPTURE-3C-seq 绘制 β-珠蛋白位点的相互作用，以高分辨率绘制染色质相互作用图目标 3 将探讨本质上的相分离现象的功能作用。 BCL11A 的无序区域 (IDR) 是最近发现的一个范例。改变生物分子在体内如何发挥其功能的观点我的研究将集中于 BCL11A 阶段是否存在。这些研究将为 BCL11A 如何调节分离珠蛋白基因表达提供机制见解。促进球蛋白转换，这是 BCL11A 治疗疾病的靶向治疗的基础。我的长期职业目标是成为表观遗传学领域一位富有成效且有影响力的科学家为了实现这一目标，我研究了发育过程中和血液疾病中的转录基因控制。为该奖项的指导阶段制定了详细的科学和职业发展计划，其中为了提高我作为调查员和小组领导者的技能，我邀请了两位已建立的和一位。初级研究员在我的咨询委员会中任职并指导我的培训。会议和实践，主要在波士顿儿童医院和周边机构举行，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。这些机构、我的导师和咨询委员会的宝贵批评和支持以及支持 K99 奖项将极大地促进那些必要的专业技能的发展成功转型为独立调查员。