Isolation and killing of candidate chronic myeloid leukemia stem cells by antibody targeting of IL-1 receptor accessory protein
Isolation and killing of candidate chronic myeloid leukemia stem cells by antibody targeting of IL-1 receptor accessory protein
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DOI：
10.1073/pnas.1004408107
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发表时间：
2010-09-14
影响因子：
11.1
通讯作者：
Fioretos, Thoas
中科院分区：
文献类型：
--
作者：
Jaras, Marcus;Johnels, Petra;Fioretos, Thoas
慢性粒细胞白血病 (CML) 的遗传特征是费城 (Ph) 染色体，该染色体是通过 9 号和 22 号染色体之间的相互易位形成的，并产生持续活跃的酪氨酸激酶 P210 BCR/ABL1。旨在治愈 CML 的治疗策略将需要完全根除 Ph 染色体阳性 (Ph+) CML 干细胞。在这里，我们使用基因表达谱来鉴定 IL-1 受体辅助蛋白 (IL1RAP) 在 CML CD34(+) 细胞和脐带血 CD34(+) 细胞中由于逆转录病毒 BCR/ABL1 表达而上调。为了测试 IL1RAP 表达是否区分 CML CD34(+)CD38(-) 细胞室内的正常 (Ph-) 和白血病 (Ph+) 细胞，我们建立了一种独特的方案，用于对少量分选细胞进行 FISH。通过使用这种方法，我们将细胞直接分选到载玻片上的液滴中以研究它们的 Ph-染色体状态。有趣的是，我们发现CML CD34(+)CD38(-)IL1RAP(+)细胞为Ph+，而CML CD34(+)CD38(-)IL1RAP(-)细胞几乎全部为Ph-。通过对两个细胞群进行长期培养起始细胞测定，我们发现可以前瞻性地分离 Ph+ 和 Ph- 候选 CML 干细胞。此外，通过产生抗IL1RAP抗体，我们提供了IL1RAP可以用作CML CD34(+)CD38(-)细胞的靶标以诱导抗体依赖性细胞介导的细胞毒性的概念证明。因此，这项研究将 IL1RAP 确定为区分 Ph+ 和 Ph- 候选 CML 干细胞的独特细胞表面生物标志物，并开辟了一条先前未探索的 CML 治疗途径。
Chronic myeloid leukemia (CML) is genetically characterized by the Philadelphia (Ph) chromosome, formed through a reciprocal translocation between chromosomes 9 and 22 and giving rise to the constitutively active tyrosine kinase P210 BCR/ABL1. Therapeutic strategies aiming for a cure of CML will require full eradication of Ph chromosome-positive (Ph+) CML stem cells. Here we used gene-expression profiling to identify IL-1 receptor accessory protein (IL1RAP) as up-regulated in CML CD34(+) cells and also in cord blood CD34(+) cells as a consequence of retroviral BCR/ABL1 expression. To test whether IL1RAP expression distinguishes normal (Ph-) and leukemic (Ph+) cells within the CML CD34(+)CD38(-) cell compartment, we established a unique protocol for conducting FISH on small numbers of sorted cells. By using this method, we sorted cells directly into drops on slides to investigate their Ph- chromosome status. Interestingly, we found that the CML CD34(+)CD38(-)IL1RAP(+) cells were Ph+, whereas CML CD34(+)CD38(-)IL1RAP(-) cells were almost exclusively Ph-. By performing long-term culture-initiating cell assays on the two cell populations, we found that Ph+ and Ph- candidate CML stem cells could be prospectively separated. In addition, by generating an anti-IL1RAP antibody, we provide proof of concept that IL1RAP can be used as a target on CML CD34(+)CD38(-) cells to induce antibody-dependent cell-mediated cytotoxicity. This study thus identifies IL1RAP as a unique cell surface biomarker distinguishing Ph+ from Ph- candidate CML stem cells and opens up a previously unexplored avenue for therapy of CML.