Replication-incompetent gammaretroviral and lentiviral vector-based insertional mutagenesis screens identify prostate cancer progression genes.
Replication-incompetent gammaretroviral and lentiviral vector-based insertional mutagenesis screens identify prostate cancer progression genes.
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DOI：
10.18632/oncotarget.24503
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发表时间：
2018-03-20
期刊：
影响因子：
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通讯作者：
Trobridge GD
中科院分区：
文献类型：
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作者：
Bii VM;Collins CP;Hocum JD;Trobridge GD
复制无能的γ -逆转录病毒（γ - rv）和慢病毒（LV）载体都被用于插入突变筛选，以识别癌症驱动因素。在这种方法中，载体稳定地整合在宿主细胞基因组中，并通过失调附近的基因诱导癌症。在原癌基因或肿瘤抑制因子内或附近含有逆转录病毒载体原病毒的细胞在肿瘤中优先富集。γ - rv和LV载体具有不同的整合谱和基因毒性潜力，使它们成为插入突变筛选的潜在补充工具。我们在异种移植小鼠模型中使用γ - rv和LV载体进行筛选，以鉴定介导雄激素非依赖性前列腺癌（AIPC）进展的驱动基因。将载体转导的LNCaP细胞原位注入免疫缺陷小鼠前列腺。发生肿瘤的小鼠被阉割，以创造一个缺乏雄激素的环境，并对发生的转移性肿瘤进行分析。一种高通量改良基因组测序PCR （MGS-PCR）方法确定了这些转移性肿瘤中载体整合的位置。OR2A14、FER1L6、TAOK3、MAN1A2、MBNL2、SERBP1、PLEKHA2、SPTAN1、ADAMTS1、SLC30A5、ABCC1、SLC7A1和SLC25A24被确定为前列腺癌（PC）进展候选基因。TAOK3和ABCC1在PC患者中的表达预测雄激素剥夺治疗后复发的风险。我们的数据表明，γ - rv和LV载体是鉴定癌症驱动基因的互补方法，可能是有希望的潜在生物标志物和治疗靶点。
Replication-incompetent gammaretroviral (γRV) and lentiviral (LV) vectors have both been used in insertional mutagenesis screens to identify cancer drivers. In this approach the vectors stably integrate in the host cell genome and induce cancers by dysregulating nearby genes. The cells that contain a retroviral vector provirus in or near a proto-oncogene or tumor suppressor are preferentially enriched in a tumor. γRV and LV vectors have different integration profiles and genotoxic potential, making them potentially complementary tools for insertional mutagenesis screens. We performed screens using both γRV and LV vectors to identify driver genes that mediate progression of androgen-independent prostate cancer (AIPC) using a xenotransplant mouse model. Vector transduced LNCaP cells were injected orthotopically into the prostate gland of immunodeficient mice. Mice that developed tumors were castrated to create an androgen-deficient environment and metastatic tumors that developed were analyzed. A high-throughput modified genomic sequencing PCR (MGS-PCR) approach identified the positions of vector integrations in these metastatic tumors. OR2A14, FER1L6, TAOK3, MAN1A2, MBNL2, SERBP1, PLEKHA2, SPTAN1, ADAMTS1, SLC30A5, ABCC1, SLC7A1 and SLC25A24 were identified as candidate prostate cancer (PC) progression genes. TAOK3 and ABCC1 expression in PC patients predicted the risk of recurrence after androgen deprivation therapy. Our data shows that γRV and LV vectors are complementary approaches to identify cancer driver genes which may be promising potential biomarkers and therapeutic targets.