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

项目摘要/摘要 这位候选人最近一直在扩大他在固体癌症分子生物学方面的强大专业知识 接受RNA生物学、造血和白血病方面的重要技能和概念，并过渡到 独立研究人员。他的长期目标是了解谱系控制转录因子是如何 肿瘤抑制功能在正常和恶性造血中的调节并利用这一点 为预防和治疗目的调节它们的表达的理解。他的目标是 KO1的建议是确定PU.1是如何调控的，PU.1是一个关键的造血转录因子 正常和恶性造血。PU1也是急性髓系白血病(AML)的关键肿瘤抑制因子。 PU.1在髓系细胞中的表达是由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基因表达下调 由于Ure缺失导致的水平严重影响髓系细胞的发育，导致AML的发生。它 目前尚不清楚是什么规范了URE-PRPR的远程相互作用。在这项研究中，候选人将决定 增强子RNA调节PU1在正常和恶性肿瘤中表达的作用和机制 造血术。将追求以下具体目标：(1)鉴定增强子RNA诱导PU1 表达，对正常的造血很重要，(2)确定它诱导PU1的机制 在染色质水平上的表达，以及(3)确定它通过恢复适当的 PU.1基因座上的染色质结构，从而导致PU.1重新激活。完成这些目标将 为非编码RNA在正常和恶性造血中的核功能提供新的见解， 为通过恢复适当的染色质结构来重新激活抑癌基因提供了科学依据。 最终，这有望导致新的分子治疗方法的发展 AML。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 利用转基因、基因敲除和植入小鼠模型，通过染色质结构进行调控。在.期间 在获奖期间，候选人将专注于获得与这些拟议方法相关的新技能， 建立他的专业知识和概念性知识的基因调控的ncRNA在造血和 白血病，并获得独立的RO1型赠款。这位候选人组建了一支强大的导师团队。 他的主要导师丹尼尔·特宁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 进步。此外，他在贝丝以色列女执事那里拥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， 成功完成他提议的学业所需的动机、指导和学术环境 并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.