Structure and Function of Stability-Enhanced Beta-Globin mRNAs

稳定性增强的 β-珠蛋白 mRNA 的结构和功能

• 批准号: 8449494
• 负责人: Osheiza Y Abdulmalik
• 金额: $ 13.93万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8449494
• 关键词: 3' Untranslated Regions; Advisory Committees; Affect; Binding; Binding Sites; Cells; Commit; Cultured Cells; Data; Development Plans; Disease; Elements; Engineering; Environment; Erythroid; Erythroid Cells; Foundations; Funding; Genes; Genetic Variation; Globin; Goals; Hela Cells; Hematological Disease; Hematology; Hemoglobin; Hemoglobinopathies; Hereditary Disease; Higher Order Chromatin Structure; Human; Hybrids; Inherited; Investigation; Journals; K-562; K562 Cells; Knowledge; Length; Maps; Medicine; Mentors; Messenger RNA; Modification; Molecular; Morbidity - disease rate; Pediatric Hospitals; Pennsylvania; Phenotype; Philadelphia; Pilot Projects; Poly(A) Tail; Population; Proteins; Public Health; Research; Research Personnel; Sickle Cell Anemia; Structure; System; Techniques; Testing; Thalassemia; Therapeutic; Training; Trans-Activators; Transgenes; Translating; Translations; United States; Universities; Untranslated Regions; Variant; Work; abstracting; base; beta Globin; beta Thalassemia; career; career development; clinical application; design; experience; functional genomics; in vivo; insight; mRNA Stability; mortality; novel; novel strategies; novel therapeutic intervention; public health relevance; skills; stem; success; therapeutic protein; therapeutic transgene

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.

描述（由申请人提供）：该项目的总体目标是为Osheiza Abdulmalik博士提供强烈指导的科学和知识培训，这对于他过渡到分子血液学领域的独立研究人员至关重要。这项培训将由他的主要导师J. Eric Russell博士，凯瑟琳·高中博士以及经验丰富的咨询委员会的熟练调查委员会熟练地指导。在费城儿童医院和宾夕法尼亚大学提供的大量支持机制将促进培训，并提供丰富的支持机制。拟议的工作是申请人最近项目的直接扩展，并专注于负责血红蛋白疾病的基因变种，这些变体是全球最普遍的遗传性疾病之一。长期目标是揭示负责人β-珠蛋白mRNA基线稳定性的机制，并发现操纵转基因以增强稳定性编码mRNA的方法。预计这些高度稳定的mRNA将积聚到高水平，并将大量的血液疾病治疗蛋白转化为血液疾病，重点是镰状细胞疾病和β-核阿无血症。三个特定的目标旨在确认和扩展大量相关的初步数据，并为Abdulmalik博士提供一个框架，以发展技能，这对于他作为独立研究者的最终成功至关重要。他的试点数据表明，通过在其3'未翻译区域内复制干循环结构（3'UTR），可以在培养细胞中增强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）研究和评估在稳定转染的培养的红细胞中体内变异的人β-珠蛋白mRNA的稳定性； 2）建立mRNA稳定性的自主函数增强了红细胞和非果蝇细胞中的β-珠蛋白3'UTR，因为自主运作的mRNA稳定元件将对其他治疗基因构成巨大的额外价值； 3）研究双茎环β-珠蛋白3'UTR的关键结构特征，该特征可能暗示了其mRNA稳定活性的机制。拟议的研究工作，结合了教学培训和结构化指导，将为操纵mRNA稳定性开发新的血红蛋白疾病疗法提供新的见解，并将启用P.I.确立自己是一名成功且高产的独立研究人员。