Retrovirus-mediated gene transfer of NPM-ALK causes lymphoid malignancy in mice
Retrovirus-mediated gene transfer of NPM-ALK causes lymphoid malignancy in mice
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DOI：
10.1182/blood.v90.8.2901
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发表时间：
1997-10-15
期刊：
影响因子：
20.3
通讯作者：
Morris, SW
中科院分区：
文献类型：
--
作者：
Kuefer, MU;Look, AT;Morris, SW
大约5%到10%的非霍奇金淋巴瘤包含t(2;5)（p23;q35）染色体重排，我们之前已经证明这导致融合蛋白核磷蛋白间变性淋巴瘤激酶（NPM-ALK）的产生。为了评估NPM-ALK在动物模型中的转化潜力，我们用逆转录病毒储存液感染5-氟尿嘧啶处理过的小鼠骨髓，并将感染的骨髓移植到经致死照射的BALB/cByJ小鼠体内。雄性小鼠在10周龄时被移植了来自雌性供体的骨髓，其中7只小鼠接受了含有人类NPM-ALK cDNA的逆转录病毒构建物pSR α MSVtkneo-NPM-ALK的骨髓，4只作为对照组，接受了“空”pSR α msvtkneo感染的骨髓。而对照组的所有小鼠在移植后11个月都活得很好，移植含有NPM-ALK构建体的骨髓的7只小鼠中有4只在4至6个月内发生淋巴瘤。肿瘤出现在肠系膜淋巴结，并转移到肺、肾、肝、脾和棘旁区。当将肿瘤细胞和骨髓细胞移植到亚致死照射的二次受体中时，这13只小鼠中有10只在9个月内发生肿瘤。使用ALK多克隆抗体对细胞裂解物进行免疫印迹分析，结果显示NPM-ALK在所有肿瘤中均有表达。组织学上，肿瘤由大的具有嗜碱性细胞质、细胞核位于中心、核仁明显的免疫母细胞组成。基因型分析显示，肿瘤为b系和克隆型，Ig重链和kappa轻链位点重排，t细胞受体β位点无重排。免疫细胞化学研究证实肿瘤细胞内存在IgM重链和kappa轻链。因此，在这种逆转录病毒基因转移模型中，NPM-ALK在小鼠中的表达导致b系大细胞淋巴瘤，这表明这种激活的融合酪氨酸激酶在人淋巴瘤中起直接的致病作用。(C) 1997年由美国血液病学会出版。
Approximately 5% to 10% of all non-Hodgkin's lymphomas contain a t(2;5)(p23;q35) chromosomal rearrangement, which we have previously shown results in the generation of the fusion protein nucleophosmin-anaplastic lymphoma kinase (NPM-ALK). To assess the transforming potential of NPM-ALK in an animal model, we infected 5-fluorouracil-treated murine bone marrow using retroviral stocks and transplanted this infected marrow into lethally irradiated BALB/cByJ mice. Male mice were transplanted with bone marrow from female donors at 10 weeks of age, with 7 of the animals receiving marrow infected with a retroviral construct, pSR alpha MSVtkneo-NPM-ALK, that contains the human NPM-ALK cDNA, and 4 serving as a control group, receiving ''empty'' pSR alpha MSVtkneo-infected marrow. Whereas all mice in the control group were alive and well up to 11 months after transplantation, 4 of the 7 mice transplanted with marrow containing the NPM-ALK construct developed lymphoma within 4 to 6 months. Tumors arose in the mesenteric lymph nodes, with metastases to the lungs, kidneys, liver, spleen, and the paraspinal area. When cells from the tumors and bone marrow were transplanted into sublethally irradiated secondary recipients, 10 of these 13 mice developed tumors within 9 months. Immunoblot analysis of cell lysates using an ALK polyclonal antibody showed NPM-ALK expression in all tumors examined. Histologically, the tumors were composed of a uniform population of large immunoblastic cells with basophilic cytoplasm, centrally placed nuclei, and distinct nucleoli. Genotypic analysis showed that the tumors were B-lineage and clonal, with rearrangements of the Ig heavy-and kappa light-chain loci and no rearrangements of the T-cell receptor beta locus. Immunocytochemical studies confirmed the presence of IgM heavy chains and kappa light chains within the tumor cells. Thus, in this retroviral gene transfer model, NPM-ALK expression in mice causes B-lineage large-cell lymphoma, suggesting a direct causative role for this activated fusion tyrosine kinase in human lymphoma. (C) 1997 by The American Society of Hematology.