Functional Analysis of RPL35A Alterations in Diamond Blackfan Anemia

描述(由申请者提供)：这份建议书概述了一项为期5年的儿科血液学/肿瘤学学术研究发展计划。这位首席研究员建议通过重点研究钻石-黑扇贫血(DBA)的新发展途径，进一步积累他在遗传性骨髓衰竭综合征机制方面的研究经验和专业知识。钻石-黑扇贫血是一种骨髓衰竭综合征，破坏红细胞穿透，导致先天性异常，是一种癌症易感综合征。这一研究计划将约翰霍普金斯大学广泛的部门和核心设施资源与国家注册中心的临床资源相结合，为在该领域发展积极和富有成效的职业生涯提供了一个极好的环境。罗伯特·阿塞奇博士是儿童血液学和肿瘤学领域的领军人物，在白血病的髓系发育异常和分子研究方面拥有专长，他将赞助并担任首席导师。与北美钻石黑粉贫血登记处(DEAR)的合作将确保获得关键患者样本和临床数据。一个由骨髓衰竭、DBA和核糖体生物发生领域的领导人组成的咨询委员会将提供额外的科学和职业指导。本研究将通过对RPL35a的研究来了解核糖体蛋白异常在DBA发生中的作用。RPL35a是我们实验室发现的一种新的DBA的核糖体亚基蛋白。其具体目的是：1)通过突变测序和比较基因组杂交分析，描述核糖体蛋白突变的类型和频率及其相关的临床特征，以确定缺失；2)使用可诱导的慢病毒siRNA模型，在造血细胞系和骨髓中确定RPL35a异常对造血的细胞后果；以及3)通过分析核糖体组装/mRNA募集和蛋白质表达变化，确定RPL35a异常的分子后果。相关性(请参阅说明)：。通过仔细定义钻石黑粉贫血的新遗传原因，这些研究将提高诊断和向高危家庭提供遗传咨询的能力，并可能为更有效的治疗确定新的靶点。该项目还将在进一步确定蛋白质合成异常与骨髓衰竭、出生缺陷和癌症易感性之间的联系方面具有更广泛的相关性。

DESCRIPTION (provided by applicant): This proposal outlines a 5-year program for the development of an academic research career in pediatric hematology/oncology. The principal investigator proposes to further his research experience and develop expertise in the mechanisms of inherited bone marrow failure syndromes through focused investigation of a novel pathway for the development of Diamond-Blackfan Anemia (DBA), a bone marrow failure syndrome that disrupts erythropoesis, leads to congenital abnormalities, and is a cancer-predisposition syndrome. This research program integrates extensive departmental and core facility resources at Johns Hopkins with the clinical resources of a national registry to provide a superb environment from which to develop an active and productive career in this field. Dr. Robert Arceci, a leader in pediatric hematology and oncology with expertise in abnormal myeloid development and molecular studies in leukemia, will sponsor and serve as chief mentor. Collaboration with the Diamond Blackfan Anemia Registry of North America (DEAR) will ensure access to critical patient samples and clinical data. An advisory committee of leaders in the fields of bone marrow failure, DBA and ribosome biogenesis will provide additional scientific and career mentoring. The research will focus on understanding the contribution of ribosomal protein abnormalities to the development of DBA through investigation of RPL35a, a large ribosomal subunit protein our laboratory identified as a novel cause of DBA. The specific aims are to: 1) delineate the types and frequency of ribosomal protein mutations and their associated clinical characteristics by sequencing for mutations and comparative genomic hybridization analysis to identify deletions, 2) determine the cellular consequences of RPL35a abnormalities to hematopoesis using an inducible lentiviral siRNA model in hematopoietic cell lines and bone marrow, and 3) determine the molecular consequences of RPL35a abnormalities by analysis of ribosome assembly/mRNA recruitment and protein expression changes. RELEVANCE (See instructions): . By carefully defining a new genetic cause of Diamond Blackfan anemia, these studies will improve the ability to diagnose and provide genetic counseling to at-risk families and may identify new targets for more effective treatments. This project will also have broader relevance in further defining the link between abnormalities of protein synthesis and bone marrow failure, birth defects, and cancer predisposition.