Roles of BCL11A in γ-globin repression

BCL11A 在 γ-珠蛋白抑制中的作用

• 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
• 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

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.

项目摘要 胎儿γ-珠蛋白如何在成人红系细胞中被关闭是珠蛋白开关的中心问题。更清楚 对潜在机制的理解构成了胎儿血红蛋白（HbF）临床再活化的基础。 一种治疗β-血红蛋白疾病如镰状细胞病和β-地中海贫血的治疗方法， 世界上最流行的遗传病。以前的研究已经确定了BCL 11 A作为一种主要的 γ-珠蛋白的阻遏物，但其机制仍不清楚。本研究的重点是机械解剖， BCL 11 A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。 我的广泛和长期目标是提高我们对发育基因控制的基本理解， 从而加速药物发现以造福公众健康。 在指导阶段，BCL 11 A作用的详细机制将使用强大的新功能进行研究。 发达的技术。目的1询问BCL 11 A如何与γ-球蛋白的转录激活因子竞争 发起人。SMASh-BCL 11 A降解系统的使用使得能够可逆地控制BCL 11 A降解， 这与基因敲除相反，在基因敲除中，人们只能在长时间的间隔后检查损失的影响。 CUT&RUN将用于绘制BCL 11 A和转录激活因子结合的高分辨率图， 控制BCL 11 A降解和再表达。Aim 2检测BCL 11 A在组织染色质中的作用 使用CAPTURE-3C-seq在β-珠蛋白基因座处的相互作用，其以高分辨率绘制染色质相互作用 和敏感性。目的3将探讨内禀相分离现象的功能作用 BCL 11 A的无序区域（IDR）。相分离是最近的一个发现， 改变了人们对生物分子如何在体内发挥作用的看法。我的研究将集中在BCL 11 A阶段是否 分离调节珠蛋白基因表达。这些研究将提供有关BCL 11 A如何作用的机制见解 促进珠蛋白转换，这是BCL 11 A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有影响力的科学家， 在发育和血液疾病中的转录基因控制。为了实现这一目标，我 为该奖项的指导阶段制定了详细的科学和职业发展计划， 旨在提高我作为调查员和组长的技能我邀请了两个知名人士和一个 初级调查员在我的咨询委员会任职，指导我的培训。拟议的活动包括课程， 会议和实践，主要发生在波士顿儿童医院和周边机构， 包括哈佛医学院和丹娜-法伯癌症研究所。一流的培训环境提供 这些机构，我的导师和咨询委员会的宝贵批评和支持， 从这个K99奖将大大推进这些必要的专业技能的发展，使一个 成功转型为独立调查员