Novel therapeutic targets for Diamond Blackfan anemia and other ribosomopathies

描述（由申请人提供）：候选人Anupama Narla提出了一个5年的职业发展计划，旨在利用三种不同的方法，寻找治疗Diamond Blackfan贫血（DBA）和其他核糖体功能障碍的新药物，同时建立一个转化儿科血液学的学术生涯。本研究的具体目的是：(1)确定来那度胺对红细胞分化的作用机制，研究来那度胺治疗钻石黑扇贫血的潜在作用；(2)生成并鉴定来自Diamond Blackfan贫血和5q-综合征患者的iPS细胞；(3)在核糖体单倍不全的细胞中进行RNAi筛选，以确定新的治疗药物。她的主要导师本杰明·艾伯特（Benjamin Ebert）博士将RPS14确定为5q-综合征红系表型的候选基因，并迅速确立了自己在新兴核糖体疾病领域的领导地位。最近才在人类中发现的核糖体病包括一系列疾病，其中遗传异常导致核糖体生物发生和功能受损。除了DBA和5q-综合征外，正常核糖体生物发生所需的其他基因突变与几种罕见的先天性综合征有关，包括Shwachman-Diamond综合征、先天性角化不良症、软骨毛发发育不全和Treacher Collins综合征。在相对较短的时间内，候选人在Ebert博士的直接监督下，熟练掌握了成功实现其提案目标所需的所有技术和方法技能。她描述了地塞米松和来那度胺单独和联合使用对体外人骨髓原代祖细胞分化的影响。她发现这两种药物都能促进正常CD34+细胞和DBA和del（5q）骨髓增生异常综合征中核糖体功能障碍类型的CD34+细胞的红细胞生成。她还与Broad研究所合作，利用细胞培养中氨基酸的稳定同位素标记（SILAC）确定了CBR1作为来那度胺的靶标。此外，她还从一位DBA患者身上培育出了红细胞生成缺陷的iPS细胞系。她的第一作者论文关于地塞米松和来那度胺对红细胞生成的影响，在首轮积极评价后，于2011年3月重新提交给Blood。她是核糖体疾病领域的三篇综述论文的第一作者，这些论文已经或正在发表在Blood， International Journal of Hematology和Seminars in Hematology上。她是一篇论文的第二作者，该论文描述了p53激活在核糖体单倍不全中对红细胞的特异性影响；这篇论文作为《血液》印刷版的一篇全体会议文章。此外，另一篇第二作者的论文确定了microRNA miR-145在5q-综合征病理生理中的作用，将于2011年5月再次提交给Blood。2009年12月，她在美国血液学学会的口头会议上介绍了这项工作。基于她在Ebert实验室的科学成就和她在儿科血液学方面的背景，候选人概述了她的短期和长期职业目标，她将继续将80%的精力投入到转化血液学研究中，20%的时间用于骨髓衰竭综合征的临床实践和教育。为了实现这些目标，她组建了一支由导师、顾问和合作者组成的才华横溢的团队。除了她的主要导师Ebert博士之外，候选人还将由Leonard Zon共同指导，Leonard Zon在成功指导医师科学家方面有着良好的记录。她的导师将至少每月与候选人会面一次，以监督和帮助她过渡到独立。David Williams和Nancy Berliner都是血液学的部门主管和国际领导者，他们将担任候选人的研究委员会，该委员会将至少每六个月召开一次会议，以确保科学研究计划的成功。该候选人还聘请了ip技术领域的领导者George Daley；Colin Sieff，波士顿儿童医院骨髓衰竭诊所主任；以及Steven Carr，他领导了基于silac的目标识别技术的发展，该技术允许对与药物结合的蛋白质进行公正的识别，作为她项目的合作者。最后，候选人可以在波士顿儿童医院的卡普研究大楼进行她的研究，该大楼还拥有大卫·威廉姆斯、斯科特·阿姆斯特朗、斯图尔特·奥金、伦纳德·佐恩、南希·柏林和乔治·戴利的实验室。这种对造血的临床和科学专业知识的集中将成为Narla博士的额外资源。执行本提案中概述的具体目标和研究职业发展计划将使候选人成功过渡到专注于开发核糖体生物发生障碍新疗法的转化研究的独立研究者的职业生涯。

DESCRIPTION (provided by applicant): The candidate, Anupama Narla, presents a 5-year career development plan that seeks to identify novel therapeutic agents for Diamond Blackfan anemia (DBA) and other disorders of ribosome dysfunction, using three different approaches, while establishing an academic career in translational pediatric hematology. The specific aims of this proposal are to: (1) determine the mechanism of action of lenalidomide on erythroid differentiation and study its potential role in the treatment of Diamond Blackfan anemia; (2) generate and characterize iPS cells derived from patients with Diamond Blackfan anemia and the 5q- syndrome; and (3) perform an RNAi screen in cells with ribosomal haploinsufficiency in order to identify novel therapeutic agents. Her primary mentor, Dr. Benjamin Ebert, identified RPS14 as the candidate gene for the erythroid phenotype of the 5q- syndrome, and has quickly established himself as a leader in the emerging field of ribosomopathies. Ribosomopathies, which have only very recently been identified in humans, comprise a collection of disorders in which genetic abnormalities cause impaired ribosome biogenesis and function. In addition to DBA and the 5q- syndrome, mutations in other genes required for normal ribosome biogenesis have been implicated in several rare congenital syndromes including Shwachman-Diamond Syndrome, Dyskeratosis Congenita, Cartilage Hair Hypoplasia, and Treacher Collins Syndrome. In a relatively short period of time, the candidate, under the direct supervision of Dr. Ebert, has gained proficiency in all of the technical and methodological skills needed to successfully carry out the objectives of her proposal. She has characterized the effects of dexamethasone and lenalidomide, individually and in combination, on the differentiation of primary human bone marrow progenitor cells in vitro. She found that both agents promote erythropoiesis in normal CD34+ cells and in CD34+ cells with the types of ribosome dysfunction found in DBA and del(5q) myelodysplastic syndrome. She has also identified CBR1 as a target of lenalidomide using stable isotope labeling with amino acids in cell culture (SILAC) in collaboration with the Broad Institute. And she has been instrumental in the generation of an iPS cell line with defective erythropoiesis, derived from a patient with DBA. Her first author paper on the effects of dexamethasone and lenalidomide on erythropoiesis is being re-submitted to Blood in March 2011 after an initial round of positive reviews. She is the first author on three review papers in the field of ribosomopathies which have been or are being published in Blood, International Journal of Hematology, and Seminars in Hematology. She is the second author on a paper that described the erythroid specific effects of p53 activation in ribosomal haploinsufficiency; this paper was featured as a plenary article in the print edition of Blood. In addition, another second author paper which identified the role of a microRNA, miR-145, in the pathophysiology of the 5q- syndrome will be resubmitted to Blood in May 2011. She presented this work as an oral session at the American Society of Hematology in December 2009. Building on her scientific accomplishments in the Ebert lab and her background in pediatric hematology, the candidate has outlined both short and long-term career goals in which she would continue to devote 80% of her effort to research in translational hematology and 20% of her time to clinical practice in bone marrow failure syndromes and education. To accomplish these goals, she has assembled a highly talented team of mentors, advisors, and collaborators. In addition to her primary mentor, Dr. Ebert, the candidate will be co-mentored by Leonard Zon who has an established record of successfully mentoring physician-scientists. Her mentors will meet with the candidate at least monthly to supervise and assist in her transition to independence. David Williams and Nancy Berliner, both division heads and international leaders in hematology, are serving as the candidate's research committee which will meet, at minimum, every six months to ensure a successful scientific research program. The candidate has also recruited George Daley, a leader in the field of iPS technology; Colin Sieff, Director of the Bone Marrow Failure Clinic at Children's Hospital Boston; and Steven Carr, who led the development of SILAC-based target identification which allows for the unbiased identification of proteins that bind to a drug, as collaborators for her project. Finally, the candidate has the benefit of carrying out her proposed research in the Karp Research Building at Children's Hospital Boston which also houses the labs of David Williams, Scott Armstrong, Stuart Orkin, Leonard Zon, Nancy Berliner, and George Daley. This concentration of clinical and scientific expertise on hematopoiesis will be an additional resource to Dr. Narla. Execution of the specific aims and research career development plan outlined in this proposal will allow the candidate to successfully transition to a career as an independent investigator in translational research focused on developing new therapies for disorders of ribosome biogenesis.