Single cell RNA-seq analysis of the effects of idasanutlin and/or nilotinib on primary human CML cells engrafted into the bone marrow of immunocompromised mice

Single cell RNA-seq was performed on primary human CML cells engrafted into the bone marrow of immunocompromised mice to understand the molecular consequences of treatment with the tyrosine kinase inhibitor nilotinib alone and in combination with the MDM2 inhibitor idasanutlin. CML CD34+ cells were engrafted into the BM of irradiated immuno-compromised mice (cohort sizes 3 mice per arm per experiment). Eight to twelve weeks post-transplantation, mice were treated with nilotinib (NIL) (50 mg/kg QD) for 7-14 days after which the cohorts received NIL for a further 14 days +/- IDASA (150 mg/kg BD) for the first 5 days. At the end of 28 day treatment or after a further 28 days treatment-free period mice were analyzed for human cell engraftment in the murine bone marrow and scRNA-seq was performed on huCD45+ or huCD34+ cells.

为探究酪氨酸激酶抑制剂（tyrosine kinase inhibitor）单药及与MDM2抑制剂（MDM2 inhibitor）艾达诺林（idasanutlin）联合治疗的分子效应，本研究将原发性人类慢性髓系白血病（Chronic Myeloid Leukemia, CML）细胞移植至免疫缺陷小鼠骨髓中，开展单细胞RNA测序（single cell RNA-seq）。
具体实验设置为：将CML来源的CD34阳性细胞移植至经辐照处理的免疫缺陷小鼠骨髓（bone marrow, BM）中；每轮实验中，每个实验臂设置3只小鼠作为实验队列。移植后8至12周，小鼠先以尼洛替尼（nilotinib, NIL）50 mg/kg 每日一次（QD）给药7至14天；随后部分实验组继续予以尼洛替尼治疗14天，其中前5天联合给予艾达诺林150 mg/kg 每日两次（BD）。在28天治疗周期结束时，或再经过28天无治疗恢复期后，对小鼠进行检测以分析其骨髓中人源细胞的嵌合情况，并对人源CD45阳性（huCD45+）或人源CD34阳性（huCD34+）细胞开展单细胞RNA测序。