Effective drug treatment identified by in vivo screening in a transplantable patient-derived xenograft model of chronic myelomonocytic leukemia
Effective drug treatment identified by in vivo screening in a transplantable patient-derived xenograft model of chronic myelomonocytic leukemia
复制标题
DOI：
10.1038/s41375-020-0929-3
复制
发表时间：
2020-06-24
期刊：
影响因子：
11.4
通讯作者：
Heuser, Michael
中科院分区：
文献类型：
--
作者：
Kloos, Arnold;Mintzas, Konstantinos;Heuser, Michael
为了建立新的和有效的治疗慢性粒单核细胞白血病(CMML)的临床前治疗组合，我们假设CMML细胞与人致癌脑膜瘤1(MN1)的补充促进了小鼠的扩增和连续移植，同时保持了这些细胞对其原始遗传特征的功能依赖。利用慢病毒表达的MN1作为致癌基因的补充，并将转导的原代单个核CMML细胞移植到免疫低下的小鼠体内，我们建立了三种可连续移植的CMML-PDX模型，其疾病相关基因突变在体内重现了疾病。异位MN1的表达证实可以促进CMML细胞的增殖，否则在二次移植时不会植入。此外，添加了MN1的CMML细胞可以连续移植到受体小鼠体内，最多可以移植到5代。这种强大的植入使针对CMML相关突变基因(包括NRAS)的体内RNA干扰筛选成为可能，证实它们的功能相关性在MN1存在的情况下保持不变。与氮胞苷和MEK抑制剂曲美替尼的新联合治疗相加地抑制ERK的磷酸化，从而耗尽突变的NRAS的信号。与单一药物治疗相比，联合治疗显著延长了CMML小鼠的存活时间。因此，我们确定阿扎替丁和曲美替尼联合治疗NRAS突变的CMML是有效的，并提出了其临床发展方向。
To establish novel and effective treatment combinations for chronic myelomonocytic leukemia (CMML) preclinically, we hypothesized that supplementation of CMML cells with the human oncogeneMeningioma 1 (MN1)promotes expansion and serial transplantability in mice, while maintaining the functional dependencies of these cells on their original genetic profile. Using lentiviral expression of MN1 for oncogenic supplementation and transplanting transduced primary mononuclear CMML cells into immunocompromised mice, we established three serially transplantable CMML-PDX models with disease-related gene mutations that recapitulate the disease in vivo. Ectopic MN1 expression was confirmed to enhance the proliferation of CMML cells, which otherwise did not engraft upon secondary transplantation. Furthermore, MN1-supplemented CMML cells were serially transplantable into recipient mice up to 5 generations. This robust engraftment enabled an in vivo RNA interference screening targeting CMML-related mutated genes includingNRAS, confirming that their functional relevance is preserved in the presence of MN1. The novel combination treatment with azacitidine and the MEK-inhibitor trametinib additively inhibited ERK-phosphorylation and thus depleted the signal from mutated NRAS. The combination treatment significantly prolonged survival of CMML mice compared to single-agent treatment. Thus, we identified the combination of azacitidine and trametinib as an effective treatment in NRAS-mutated CMML and propose its clinical development.