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: 8106161
• 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/8106161
• 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. 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)

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和智力培训，这对于他过渡到分子血液学领域的独立研究员至关重要。该培训将由他的主要导师 J. Eric Russell 博士、共同导师 Katherine High 博士以及由致力于培训年轻研究人员的经验丰富的研究人员组成的咨询委员会提供专业指导。费城儿童医院和宾夕法尼亚大学活跃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，重点关注导致血红蛋白疾病的基因变异，血红蛋白疾病是全世界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，并发现操纵转基因编码具有增强稳定性的 mRNA 的方法。这些高度稳定的 mRNA 预计会积累到高水平，并转化大量治疗血液疾病的蛋白质，重点是镰状细胞病和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，通过在 3' 非翻译区 (3'UTR) 内复制茎环结构，可以增强培养细胞中 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）研究和评估稳定转染的培养红系细胞中变异人β-珠蛋白mRNA的体内稳定性； 2) 在红系和非红系细胞中建立增强β-珠蛋白3'UTRs的mRNA稳定性的自主功能，因为自主功能的mRNA稳定元件将为其他治疗性转基因带来巨大的附加价值； 3) 研究双茎环 β-珠蛋白 3'UTR 的关键结构特征，这可能表明其 mRNA 稳定活性的机制。拟议的研究工作与教学培训和结构化指导相结合，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，并使 P.I.使自己成为一名成功且富有成效的独立资助研究员。 公共卫生相关性：血红蛋白异常会导致世界范围内一些最普遍的遗传性疾病。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造成显着的发病率和死亡率。我们提出的研究旨在调查受影响人群并开发新的治疗方案，这与公共卫生直接相关。 （摘要完）