Novel therapeutic targets for Diamond Blackfan anemia and other ribosomopathies

描述（由申请人提供）：候选人Anupama Narla提出了一项为期5年的职业发展计划，旨在使用三种不同的方法确定Diamond Blackfan贫血（DBA）和其他核糖体功能障碍疾病的新型治疗药物，同时建立翻译儿科血液学的学术生涯。该提案的具体目的是：（1）确定来那度胺对红细胞分化的作用机制，并研究其在治疗Diamond Blackfan贫血中的潜在作用;（2）生成并表征来自Diamond Blackfan贫血和5 q综合征患者的iPS细胞;（3）在核糖体单倍不足的细胞中进行RNAi筛选，以鉴定新型治疗药物。她的主要导师Benjamin Ebert博士将RPS 14确定为5 q综合征红细胞表型的候选基因，并迅速确立了自己在核糖体病新兴领域的领导地位。核糖体病是最近才在人类中发现的疾病，包括一系列遗传异常导致核糖体生物发生和功能受损的疾病。除了DBA和5 q-综合征之外，正常核糖体生物发生所需的其他基因的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、腕骨毛发发育不全和Treacher柯林斯综合征。在相对较短的时间内，候选人在埃伯特博士的直接监督下，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对体外原代人骨髓祖细胞分化的影响。她发现，这两种药物促进正常CD 34+细胞和DBA和del（5 q）骨髓增生异常综合征中发现的核糖体功能障碍类型的CD 34+细胞中的红细胞生成。她还与布罗德研究所合作，使用细胞培养中氨基酸的稳定同位素标记（SILAC）将CBR 1确定为来那度胺的靶点。她还帮助从DBA患者中产生了红细胞生成缺陷的iPS细胞系。她关于地塞米松和来那度胺对红细胞生成的影响的第一作者论文在第一轮积极评价后于2011年3月重新提交给Blood。她是核糖体病领域三篇综述论文的第一作者，这些论文已经或正在发表在《血液》、《国际血液学杂志》和《血液学研讨会》上。她是第二作者的论文，描述了p53激活在核糖体单倍不足的红细胞特异性作用;这篇论文是精选的血液印刷版的全体文章。此外，另一篇第二作者的论文将于2011年5月重新提交给Blood，该论文确定了microRNA（miR-145）在5 q综合征病理生理学中的作用。2009年12月，她在美国血液学会的一次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，候选人概述了短期和长期的职业目标，她将继续将80%的精力投入到转化血液学研究中，并将20%的时间用于骨髓衰竭综合征和教育的临床实践。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，埃伯特博士，候选人将共同指导伦纳德Zon谁拥有成功的指导医生科学家的既定记录。她的导师将至少每月与候选人会面，监督和协助她向独立过渡。大卫威廉姆斯和南希柏林，无论是部门负责人和国际领导人在血液学，是作为候选人的研究委员会，将举行会议，至少每六个月，以确保一个成功的科学研究计划。这位候选人还招募了iPS技术领域的领导者乔治戴利;波士顿儿童医院骨髓衰竭诊所主任科林谢夫;以及领导开发基于SILAC的靶向鉴定的史蒂文卡尔，该靶向鉴定允许无偏见地鉴定与药物结合的蛋白质，作为她的项目的合作者。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提出的研究，该大楼还设有大卫威廉姆斯、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。造血方面的临床和科学专业知识将成为Narla博士的额外资源。本提案中概述的具体目标和研究职业发展计划的执行将使候选人成功过渡到翻译研究的独立研究者职业生涯，专注于开发核糖体生物合成疾病的新疗法。

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.