Structure and Function of Stability-Enhanced Beta-Globin mRNAs

描述(由申请者提供)：该项目的总体目标是为Osheiza Abdulmalik博士提供强有力的指导科学和智力培训，这对他向分子血液学领域的独立研究人员过渡至关重要。这项培训将由他的主要导师J.Eric Russell博士、共同导师Katherine High博士以及致力于培训年轻调查人员的经验丰富的资深调查人员咨询委员会熟练指导。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助机制将为培训提供便利。拟议的工作是申请人最近项目的直接延伸，重点是导致血红蛋白紊乱的基因变异，这是世界上最常见的遗传性疾病之一。长期目标是揭开导致人类β-珠蛋白mRNA基线稳定的机制(S)，并找到操纵转基因以编码具有更高稳定性的mRNA的方法。这些高度稳定的mRNAs预计会积累到高水平，并翻译大量治疗血液疾病的治疗蛋白，重点是镰状细胞疾病和β-地中海贫血。设计了三个具体目标，以确认和扩展大量相关的初步数据，并为阿卜杜勒马利克博士提供一个发展技能的框架，这些技能对他最终成为一名独立调查人员至关重要。他的试点数据表明，在培养细胞中，通过在其3‘非翻译区(3’UTR)内复制茎环结构，可以增强β-珠蛋白mRNA的稳定性。P.I.将通过以下方式扩展这项研究：1)研究和评估不同的人β-珠蛋白mRNAs在稳定转染的红系细胞中的体内稳定性；2)建立mRNA稳定性增强的β-珠蛋白3‘UTRs在红系和非红系细胞中的自主功能，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额外价值；以及3)研究双茎环β-珠蛋白3’UTRs的关键结构特征，可能为其稳定mRNA的活性提供机制。拟议中的研究工作，结合教学培训和结构化指导，将为如何操纵mRNA稳定性以开发针对血红蛋白疾病的新疗法提供新的见解，并将使P.I.成为一名成功的、多产的独立资助的研究人员。 与公共卫生相关：血红蛋白异常导致世界上一些最普遍的遗传性遗传病。镰状细胞病和β-地中海贫血是两种严重的疾病，在美国和全球范围内继续导致显著的发病率和死亡率。我们为调查和开发受影响人群的新治疗方案而提出的研究与公共卫生直接相关。(摘要结束)

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. PUBLIC HEALTH RELEVANCE: Hemoglobins abnormalities cause some of the most prevalent inherited genetic disorders worldwide. Sickle cell disease and beta-thalassemia are two such serious disorders that continue to cause significant morbidity and mortality in the United States and globally. Our proposed research to investigate and develop new treatment options for the affected population is directly relevant to public health. (End of Abstract)