Tomasson M H, Sternberg D W, Williams I R, Carroll M, Cain D, Aster J C, Ilaria R L, Van Etten R A, Gilliland D G
Department of Medicine, Brigham and Women's Hospital, Harvard Institutes of Medicine, Boston, Massachusetts 02115, USA.
J Clin Invest. 2000 Feb;105(4):423-32. doi: 10.1172/JCI8902.
The t(5;12)(q33;p13) translocation associated with chronic myelomonocytic leukemia (CMML) generates a TEL/PDGFbetaR fusion gene. Here, we used a murine bone marrow transplant (BMT) assay to test the transforming properties of TEL/PDGFbetaR in vivo. TEL/PDGFbetaR, introduced into whole bone marrow by retroviral transduction, caused a rapidly fatal myeloproliferative disease that closely recapitulated human CMML. TEL/PDGFbetaR transplanted mice developed leukocytosis with Gr-1(+) granulocytes, splenomegaly, evidence of extramedullary hematopoiesis, and bone marrow fibrosis, but no lymphoproliferative disease. We assayed mutant forms of the TEL/PDGFbetaR fusion protein - including 8 tyrosine to phenylalanine substitutions at phosphorylated PDGFbetaR sites to which various SH2 domain-containing signaling intermediates bind - for ability to transform hematopoietic cells. All of the phenylalanine (F-) mutants tested conferred IL-3-independence to a cultured murine hematopoietic cell line, but, in the BMT assay, different F-mutants displayed distinct transforming properties. In transplanted animals, tyrosines 579/581 proved critical for the development of myeloproliferative phenotype. F-mutants with these residues mutated showed no sign of myeloproliferation but instead developed T-cell lymphomas. In summary, TEL/PDGFbetaR is necessary and sufficient to induce a myeloproliferative disease in a murine BMT model, and PDGFbetaR residues Y579/581 are required for this phenotype.
与慢性粒单核细胞白血病（CMML）相关的t(5;12)(q33;p13)易位产生了一个TEL/PDGFβR融合基因。在此，我们使用小鼠骨髓移植（BMT）试验来测试TEL/PDGFβR在体内的转化特性。通过逆转录病毒转导将TEL/PDGFβR导入全骨髓，引发了一种迅速致命的骨髓增殖性疾病，该疾病与人类CMML极为相似。移植了TEL/PDGFβR的小鼠出现白细胞增多，伴有Gr-1(+)粒细胞、脾肿大、髓外造血证据和骨髓纤维化，但无淋巴细胞增殖性疾病。我们检测了TEL/PDGFβR融合蛋白的突变形式——包括在磷酸化PDGFβR位点上的8个酪氨酸到苯丙氨酸的替换，各种含SH2结构域的信号中间体可结合到这些位点——以评估其转化造血细胞的能力。所有测试的苯丙氨酸（F-）突变体都赋予了一种培养的小鼠造血细胞系IL-3非依赖性，但在BMT试验中，不同的F-突变体表现出不同的转化特性。在移植的动物中，酪氨酸579/581被证明对骨髓增殖表型的发展至关重要。这些残基发生突变的F-突变体没有骨髓增殖的迹象，而是发展为T细胞淋巴瘤。总之，在小鼠BMT模型中，TEL/PDGFβR对于诱导骨髓增殖性疾病是必要且充分的，并且该表型需要PDGFβR残基Y579/581。
