Structure and Function of Stability-Enhanced Beta-Globin mRNAs

描述（由申请人提供）：该项目的总体目标是为Osheiza Abdulmalik博士提供强有力的科学指导和智力培训，这对他向分子血液学领域的独立研究人员过渡至关重要。这次培训将由他的主要导师J. Eric Russell博士、共同导师Katherine High博士和一个经验丰富的咨询委员会指导，该委员会由致力于培训年轻调查员的有成就的调查员组成。费城儿童医院和宾夕法尼亚大学积极的研究环境和丰富的支持机制将促进培训。拟议的工作是申请人最近项目的直接延伸，重点研究血红蛋白疾病的基因变异，这是世界上最普遍的遗传性疾病之一。长期目标是揭示人类β -珠蛋白mRNA基线稳定性的机制，并发现操纵转基因来编码具有增强稳定性的mRNA的方法。这些高度稳定的mrna有望积累到高水平，并转化大量用于血液疾病的治疗蛋白，重点是镰状细胞病和-地中海贫血。三个具体目标旨在确认和扩展大量相关的初步数据，并为Abdulmalik博士提供一个发展技能的框架，这些技能对他作为独立调查员的最终成功至关重要。他的试验数据表明，β -珠蛋白mRNA的稳定性可以通过在其3‘非翻译区（3’ utr）内复制茎环结构而在培养细胞中得到增强。P.I.将通过以下方式扩展这项研究：1)在稳定转染的培养红细胞中研究和评估变异人β -珠蛋白mrna在体内的稳定性；2)在红细胞和非红细胞中建立增强mrna稳定性的β -珠蛋白3' utr的自主功能，因为自主功能的mrna稳定元件将对其他治疗性转基因具有巨大的附加价值；3)研究双茎环β -珠蛋白3' utr的关键结构特征，这些特征可能提示其mrna稳定活性的机制。拟议的研究工作，结合教学培训和结构化指导，将为操纵mRNA稳定性的方法提供新的见解，以开发血红蛋白疾病的新疗法，并将使P.I.成为一名成功且富有成效的独立资助研究人员。

DESCRIPTION (provided by applicant): The overall goal of this project is to provide Dr. Osheiza Abdulmalik with strongly mentored scientific and intellectual training essential to his transition to an independent researcher in the field of molecular hematology. This training will be expertly guided by his primary mentor, Dr. J. Eric Russell, co-mentor Dr. Katherine High, and a highly-experienced Advisory Committee of accomplished investigators committed to training young investigators. Training will be facilitated by the active research environment and abundant support mechanisms available at the Children's Hospital of Philadelphia and the University of Pennsylvania. The proposed work is a direct extension of the applicant's recent project and focuses on gene variants responsible for hemoglobin disorders, which are among the most prevalent hereditary disorders worldwide. The long-term goal is to unravel the mechanism(s) responsible for the baseline stability of human beta-globin mRNA, and discover ways to manipulate transgenes to encode mRNAs with enhanced stabilities. These highly stable mRNAs would be expected to accumulate to high levels and translate substantial amounts of therapeutic protein for blood disorders, with emphasis on sickle cell disease and beta-thalassemia. Three specific aims are designed to confirm and extend a substantial body of relevant preliminary data, as well as provide Dr. Abdulmalik a framework for developing skills critical to his eventual success as an independent investigator. His pilot data suggest that the stability of beta-globin mRNA can be enhanced in cultured cells by duplication of a stem-loop structure within its 3' untranslated region (3'UTR). The P.I. will extend this research by: 1) investigating and assessing the stabilities of variant human beta-globin mRNAs in vivo in stably-transfected cultured erythroid cells; 2) establishing the autonomous functions of mRNA-stability enhancing beta-globin 3'UTRs in erythroid and non-erythroid cells, because autonomously functioning mRNA-stabilizing elements would hold tremendous additional value for other therapeutic transgenes; and 3) investigating key structural features of double-stem loop beta-globin 3'UTRs that may suggest a mechanism for its mRNA-stabilizing activity. The proposed research work, combined with the didactic training and structured mentoring will provide new insights into ways to manipulate mRNA stability to develop new therapies for hemoglobin disorders and will enable the P.I. to establish himself as a successful and productive independently funded researcher.