Data from: Characterization of leukemia-inducing genes using a proto-oncogene/homeobox gene retroviral human cDNA library in a mouse in vivo model

The purpose of this research is to develop a method to screen a large number of potential driver mutations of acute myeloid leukemia (AML) using a retroviral cDNA library and murine bone marrow transduction-transplantation system. As a proof-of-concept, murine bone marrow (BM) cells were transduced with a retroviral cDNA library encoding well-characterized oncogenes and homeobox genes, and the virus-transduced cells were transplanted into lethally irradiated mice. The proto-oncogenes responsible for leukemia initiation were identified by PCR amplification of cDNA inserts from genomic DNA isolated from leukemic cells. In an initial screen of ten leukemic mice, the MYC proto-oncogene was detected in all the leukemic mice. Of ten leukemic mice, 3 (30%) had MYC as the only transgene, and seven mice (70%) had additional proto-oncogene inserts. We repeated the same experiment after removing MYC-related genes from the library to characterize additional leukemia-inducing gene combinations. Our second screen using the MYC-deleted proto-oncogene library confirmed MEIS1and the HOX family as cooperating oncogenes in leukemia pathogenesis. The model system we introduced in this study will be valuable in functionally screening novel combinations of genes for leukemogenic potential in vivo, and the system will help in the discovery of new targets for leukemia therapy.

本研究旨在开发一种方法，借助逆转录病毒cDNA文库（retroviral cDNA library）与小鼠骨髓转导-移植系统，对急性髓系白血病（acute myeloid leukemia, AML）的大量潜在驱动突变开展筛选。作为概念验证，研究人员将编码已充分表征的癌基因与同源盒基因（homeobox genes）的逆转录病毒cDNA文库转导至小鼠骨髓（bone marrow, BM）细胞中，并将病毒转导后的细胞移植至致死剂量辐照的小鼠体内。通过对白血病细胞中提取的基因组DNA的cDNA插入片段进行聚合酶链式反应（PCR）扩增，可鉴定出介导白血病起始的原癌基因。在初始针对10只白血病小鼠的筛选实验中，所有受试小鼠体内均检测到MYC原癌基因。其中3只（30%）小鼠仅以MYC作为唯一转基因，剩余7只（70%）小鼠则携带额外的原癌基因插入片段。研究人员移除文库中与MYC相关的基因后重复了相同实验，以表征其他可诱导白血病发生的基因组合。使用缺失MYC的原癌基因文库开展的第二轮筛选实验证实，MEIS1与同源盒（HOX）家族基因可作为白血病发病机制中的协同致癌基因。本研究构建的模型系统可用于体内功能筛选具备致白血病潜力的新型基因组合，该系统将助力白血病治疗新靶点的发现。