ROR1 as a Therapeutic Target in Acute Lymphoblastic Leukemia

项目摘要/摘要 此申请涉及计划公告编号：PA-09-036，NIH通向独立之路 奖项(K99/R00)。 应聘者：杰弗里·泰纳博士一直在分子和细胞生物学研究领域为 过去15年暑期学生(普渡大学)，本科生研究助理(格林内尔学院)， 研究生(圣路易斯华盛顿大学)，博士后研究员(俄勒冈健康与科学 大学)。他研究过多种生物学领域，包括植物学、免疫学、微生物学、 和血液学/肿瘤学。这项研究已经在High-Impact上累计发表了21篇手稿 《自然医学》、《临床研究杂志》、《癌症研究》、《美国医学会会刊》等期刊 《美国国家科学院院刊》、《血液》以及其他许多手稿正在修改或准备中。Dr。 泰纳还在主要的国家会议上以口头陈述和海报会议的形式展示了他的作品。 泰纳博士的研究主题涉及识别目标癌基因和癌基因特异性 治疗学，例如癌症治疗可以为每个患者量身定做。为了实现这些目标， 他创造了两种筛选技术，利用siRNA或小分子激酶抑制剂来识别 功能上重要的靶基因，用于后续鉴定和基因组研究。使用这些技术， 他已经确定了一种名为ROR1的基因，该基因是所有急性淋巴细胞白血病的候选治疗靶点 有1；19号染色体易位的患者。该提案旨在描述以下机制： 该靶基因的过度表达和信号转导，并确定用于治疗的基因特异性方式 干预。泰纳博士的长期职业目标包括建立一个独立的研究实验室， 专注于癌症研究和个性化医疗。 环境：俄勒冈健康与科学大学奈特癌症研究所有165名初级教员 在不同的调查领域拥有专业知识的调查人员。泰纳博士的导师，布莱恩博士 德鲁克是奈特癌症研究所的主任。德鲁克博士在以下领域拥有20多年的经验 他从事癌症研究，并指导许多学生和研究员成为独立调查员。这 提案还包括理查德·古德曼博士、菲利普·斯特里特博士和罗伯特博士的支持声明 瑟尔斯。总体而言，这些支持科学家以及奈特癌症研究所作为一个整体拥有 用于开展本申请中提出的研究的仪器、资源和专业知识。 研究：激酶抑制剂对致癌信号通路的特异性靶向已大大改善 有多种癌症诊断的患者的临床结果，尤其是慢性髓系患者 白血病。为了将这种靶向治疗方法扩展到所有形式的癌症，致病基因必须首先 被识别和刻画。为此，我们开发了一种基于RNAi的屏幕，以快速 鉴定直接从白血病患者获得的原代癌细胞中的靶基因。使用此屏幕，我们 已经确定了一种受体酪氨酸激酶，ROR1，它在急性淋巴细胞白血病中唯一且一致地过度表达 T(1；19)阳性的急性淋巴细胞白血病(ALL)患者，约占ALL的5% 儿童ALL和成人ALL的1-2%。沉默ROR1导致细胞存活率显著降低 T(1；19)阳性的ALL细胞，但对其他儿童ALL细胞或正常白细胞的活性无影响。 此外，之前对慢性淋巴细胞白血病(CLL)的研究，它约占30% 在成人白血病病例中，已发现ROR1在大多数病例中过表达。验证ROR1作为 真正的治疗靶点需要更好地理解基因调节和 ROR1参与细胞转化的信号转导。然而，人们对此知之甚少。 调控ROR1表达的调控元件或ROR1影响的信号通路 细胞活性，目前还没有可用的策略，可以通过ROR1的治疗靶点。 在t(1；19)阳性的ALL细胞中发现ROR1的过度表达和ROR1依赖性，为 研究这个问题。我们建议多管齐下研究1)ROR1的调控 表达，2)ROR1促进转化的信号机制，3) 抑制ROR1的治疗策略的发展将有助于阐明AFP的发病机制。 ROR1依赖的恶性肿瘤，如t(1；19)阳性的ALL或CLL，并提供了新的治疗策略 对这些患者进行治疗性干预。

PROJECT SUMMARY/ABSTRACT This application addresses Program Announcement Number: PA-09-036, NIH Pathway to Independence Award (K99/R00). Candidate: Dr. Jeffrey Tyner has been training in the field of molecular and cellular biology research for the past 15 years as a summer student (Purdue University), undergraduate research assistant (Grinnell College), graduate student (Washington University in St. Louis), and post-doctoral fellow (Oregon Health & Science University). He has studied a diverse spectrum of biological fields including botany, immunology, microbiology, and hematology/oncology. This research has led to 21 cumulative published manuscripts in high-impact journals such as Nature Medicine, Journal of Clinical Investigation, Cancer Research, Proceedings of the National Academy of Science, and Blood as well as numerous other manuscripts in revision or preparation. Dr. Tyner has also presented his work in oral presentations and poster sessions at major national conferences. The theme of Dr. Tyner's research involves the identification of target oncogenes and oncogene-specific therapeutics, such that cancer therapies can be tailored to each individual patient. To accomplish these goals, he has created two screening techniques that make use of siRNA or small-molecule kinase inhibitors to identify functionally important target genes for follow-up characterization and genomic study. Using these techniques, he has identified a gene, ROR1, that is a candidate therapeutic target in all acute lymphoblastic leukemia patients with a 1;19 chromosomal translocation. This proposal aims to characterize the mechanisms of overexpression and signaling of this target gene and to identify gene-specific modalities for therapeutic intervention. Dr. Tyner's long-term career goals include establishment of an independent research lab with a focus on cancer research and personalized medicine. Environment: The Oregon Health & Science University Knight Cancer Institute has 165 primary faculty investigators who have expertise across a diverse spectrum of fields of inquiry. Dr. Tyner's mentor, Dr. Brian Druker, is the Director of the Knight Cancer Institute. Dr. Druker has over 20 years of experience in the field of cancer research and has mentored numerous students and fellows to independent investigator status. This proposal also includes statements of support from Dr. Richard Goodman, Dr. Philip Streeter, and Dr. Robert Searles. Cumulatively, these supporting scientists as well as the Knight Cancer Institute as a whole posses all the instrumentation, resources, and expertise to carry out the research proposed in this application. Research: Specific targeting of oncogenic signaling pathways with kinase inhibitors has vastly improved clinical outcomes for patients with a variety of cancer diagnoses, most notably patients with chronic myeloid leukemia. To expand this targeted-therapy approach to all forms of cancer, disease-causing genes must first be identified and characterized. Towards that end, we have developed an RNAi-based screen to rapidly identify target genes in primary cancer cells obtained directly from leukemia patients. Using this screen, we have identified a receptor tyrosine kinase, ROR1, that is uniquely and consistently overexpressed in all patients with t(1;19)-positive acute lymphoblastic leukemia (ALL), representing approximately 5% of all pediatric ALL and 1-2% of adult ALL cases. Silencing of ROR1 results in significantly decreased viability of t(1;19)-positive ALL cells, but has no effect on viability of other pediatric ALL cells or normal white blood cells. In addition, previous studies of chronic lymphocytic leukemia (CLL), which accounts for approximately 30% of adult leukemia cases, have identified ROR1 overexpression in the majority of cases. Validation of ROR1 as a bona fide therapeutic target necessitates a better understanding of the mechanisms of genetic regulation and signaling by which ROR1 contributes to cellular transformation. However, very little is known about the regulatory elements governing ROR1 expression or the signaling pathways employed by ROR1 to influence cellular viability, and there are currently no available strategies by which ROR1 can be therapeutically targeted. The finding of ROR1 overexpression and ROR1-dependence in t(1;19)-positive ALL cells offers unique tools to study this problem. We propose that a multi-pronged approach to studying 1) regulation of ROR1 expression, 2) signaling mechanisms through which ROR1 contributes to transformation, and 3) development of therapeutic strategies for inhibiting ROR1 will elucidate the disease pathogenesis of ROR1-dependent malignancies such as t(1;19)-positive ALL or CLL and offer new strategies for therapeutic intervention in these patients.