Enhancer RNA-mediated Tumor Suppressor Gene Expression in Normal and Malignant Hematopoiesis

项目总结/摘要 这位候选人最近一直在扩大他在实体癌分子生物学方面的专业知识， 接受RNA生物学，造血和白血病的重要技能和概念，并过渡到一个 独立研究员。他的长期目标是了解谱系控制转录因子是如何在转录过程中发挥作用的。 肿瘤抑制功能在正常和恶性造血中受到调节， 了解调节它们的表达以用于预防和治疗目的。他的目标是 KO 1的建议是确定PU.1，一个关键的主造血转录因子，是如何调节的， 正常和恶性造血。PU.1也是急性髓性白血病（AML）中的关键肿瘤抑制因子。 PU.1在髓样细胞中的表达是由称为PU.1的远端增强子与PU.1的远端增强子之间的相互作用诱导的。 上游调控元件（URE）和近端启动子区（PrPr）。PU.1表达减少 由于URE缺失而导致的水平降低对骨髓细胞发育具有严重影响，导致AML发展。它 目前尚不清楚是什么调节URE-PrPr的长程相互作用。在这项研究中，候选人将确定 增强子RNA调节正常和恶性肿瘤中PU.1表达的作用和机制 造血将追求以下具体目标：（1）鉴定增强子RNA诱导PU。 表达，并对正常的造血功能很重要，（2）确定其诱导PU的机制。1 在染色质水平上表达，和（3）确定它通过恢复正常的细胞增殖抑制AML进展， PU.1基因座处的染色质结构，从而导致PU.1再活化。实现这些目标将 为正常和恶性造血中非编码RNA的核功能提供了新的见解， 为通过恢复正常的染色质结构重新激活抑癌基因提供科学依据。 最终，这可能会导致新的分子治疗方法的发展， 急性髓细胞白血病这些目标将通过使用各种方法来实现，包括RNA相互作用和基因表达。 使用转基因、敲除和移植小鼠模型通过染色质结构进行调控。期间 在授予期间，候选人将专注于获得与这些提议方法相关的新技能， 建立他的专业知识和概念性知识，关于造血中ncRNA的基因调控， 白血病，并获得独立的RO 1型赠款。候选人已经组建了一个强大的导师团队。 他的主要导师，丹尼尔特嫩博士，是一位著名的造血基因调控专家， 白血病此外，外部顾问委员会将监督他的研究和职业发展 中求进工作总此外，他在贝斯以色列女执事有无与伦比的资源和专业知识， 医学中心和邻近的哈佛附属机构。因此，候选人有专业知识， 成功完成其拟议研究所需的动机、指导和学术环境 建立他的独立研究项目

PROJECT SUMMARY/ABSTRACT The candidate has recently been expanding his strong expertise in molecular biology of solid cancer to embrace important skills and concepts in RNA biology, hematopoiesis and leukemia, and to transition to an independent researcher. His long-term goals are to understand how lineage-control transcription factors having tumor suppressor functions are regulated in normal and malignant hematopoiesis and to exploit this understanding in modulating their expression for preventative and therapeutic purposes. The objective of his KO1 proposal is to determine how PU.1, a critical master hematopoietic transcription factor is regulated in normal and malignant hematopoiesis. PU.1 is also a key tumor suppressor in acute myeloid leukemia (AML). PU.1 expression in myeloid cells is induced by the interaction between the distal enhancer termed the upstream regulatory element (URE), and the proximal promoter region (PrPr). Reduction in PU.1 expression levels due to URE deletion has severe impacts on myeloid cell development leading to AML development. It remains unclear what regulates URE-PrPr long-range interaction. In this study, the candidate will determine the role and mechanism by which an enhancer RNA regulates PU.1 expression in normal and malignant hematopoiesis. The following specific aims will be pursued: (1) to identify that the enhancer RNA induces PU.1 expression and is important for normal hematopoiesis, (2) to define the mechanism by which it induces PU.1 expression at the chromatin level, and (3) to establish that it inhibits AML progression by restoring proper chromatin structure at the PU.1 locus thereby leading to PU.1 reactivation. Completion of these aims will provide new insights into the nuclear functions of noncoding RNAs in normal and malignant hematopoiesis, and provide a scientific basis for reactivating tumor suppressor genes by restoring proper chromatin structure. Ultimately, this could be expected to result in the development of novel molecular therapeutic approaches for AML. These aims will be achieved by using a variety of approaches including RNA interactions and gene regulation via chromatin structure with the use of transgenic, knockout and engraftment mouse models. During the award period, the candidate will focus on attaining new skills related to these proposed approaches, building his expertise and conceptual knowledge regarding gene regulation by ncRNAs in hematopoiesis and leukemia, and obtaining independent RO1-type grant. The candidate has assembled a strong mentor team. His primary mentor, Dr. Daniel Tenen, is a well-known expert in gene regulation of hematopoiesis and leukemia. In addition, the external advisor committee will monitor his research and career development progress. Furthermore, he has unparalleled access to resources and expertise at the Beth Israel Deaconess Medical Center and neighboring Harvard-affiliated institutions. Therefore, the candidate has the expertise, motivation, mentorship and academic environment necessary to successfully accomplish his proposed studies and build his independent research program.