Intra-heterogeneity in transcription and chemoresistant property of leukemia-initiating cells in murine Setd2(-/-) acute myeloid leukemia.
Intra-heterogeneity in transcription and chemoresistant property of leukemia-initiating cells in murine Setd2(-/-) acute myeloid leukemia.
复制标题
DOI：
10.1002/cac2.12189
复制
发表时间：
2021-09
期刊：
影响因子：
--
通讯作者：
Huang Q
中科院分区：
文献类型：
--
作者：
Song J;Du L;Liu P;Wang F;Zhang B;Xie Y;Lu J;Jin Y;Zhou Y;Lv G;Zhang J;Chen S;Chen Z;Sun X;Zhang Y;Huang Q
白血病起始细胞（LIC）的异质性是急性髓系白血病（AML）治疗的主要障碍。越来越多的证据表明，同一个体中多种不同致病性的LICs共存是AML的一个共同特征。然而，包括共存 LIC 的药物反应在内的功能异质性仍不清楚。因此，本研究旨在阐明 LIC 的内部异质性，以帮助预测白血病行为并开发更有效的治疗方法。将来自原代 Setd2−/− AML 小鼠的脾细胞移植到 C57BL/6 受体小鼠中以生成可移植模型。采用流式细胞术分析白血病小鼠的免疫表型。进行全基因组测序以检测导致白血病转化的二次命中。使用连续移植测定来确定 Setd2−/− AML 细胞的自我更新潜力。进行有限稀释测定以确定不同白血病细胞亚群中的 LIC 频率。通过批量和单细胞 RNA 测序来分析 LIC 的转录异质性。采用小分子抑制剂筛选和体内药物治疗来阐明不同 LIC 子集之间药物反应的差异。在这项研究中，我们观察到一只老年 Setd2 −/− 小鼠出现 AML，并发生 Nras G12S 和 Braf K520E 突变。进一步的研究确定了两种类型的 LIC 分别存在于 c-Kit+B220+Mac-1− 和 c-Kit+B220+Mac-1+ 子集中。体内移植测定揭示了共存 LIC 之间分化的异质性。此外，在 c-Kit+B220+Mac-1+ 细胞中发现了内在的阿霉素抗性转录特征。事实上，阿霉素加阿糖胞苷（DA）作为AML治疗中使用的标准化疗方案，可以特异性杀死c-Kit+B220+Mac-1−细胞，但几乎不影响c-Kit+B220+Mac-1+细胞。转录组分析显示，c-Kit+B220+Mac-1+ 细胞中 RAS 下游信号通路的激活程度高于 c-Kit+B220+Mac-1− 细胞。 DA 和 RAS 途径抑制剂联合治疗可杀死 c-Kit+B220+Mac-1− 和 c-Kit+B220+Mac-1+ 细胞并减弱疾病进展。这项研究鉴定了小鼠 Setd2−/− AML 中 LIC 富集的两个细胞亚群，并揭示了 LIC 的转录和功能异质性，揭示了该模型中不同类型 LIC 的共存带来了不同的药物反应。探索了小鼠 Setd2−/−−AML 中 LIC 的转录和功能异质性。该模型中两种不同类型 LIC 的共存会带来不同的化学反应。与 DA 和 RAS 通路抑制剂联合治疗可以杀死这两种 LIC 并减缓疾病进展。这些发现可能会增进我们对 LIC 内部异质性的理解，并帮助我们开发更有效的治疗方法。
Heterogeneity of leukemia‐initiating cells (LICs) is a major obstacle in acute myeloid leukemia (AML) therapy. Accumulated evidence indicates that the coexistence of multiple types of LICs with different pathogenicity in the same individual is a common feature in AML. However, the functional heterogeneity including the drug response of coexistent LICs remains unclear. Therefore, this study aimed to clarify the intra‐heterogeneity in LICs that can help predict leukemia behavior and develop more effective treatments. Spleen cells from the primary Setd2−/− ‐AML mouse were transplanted into C57BL/6 recipient mice to generate a transplantable model. Flow cytometry was used to analyze the immunophenotype of the leukemic mice. Whole‐genome sequencing was conducted to detect secondary hits responsible for leukemia transformation. A serial transplantation assay was used to determine the self‐renewal potential of Setd2−/− ‐AML cells. A limiting‐dilution assay was performed to identify the LIC frequency in different subsets of leukemia cells. Bulk and single‐cell RNA sequencing were performed to analyze the transcriptional heterogeneity of LICs. Small molecular inhibitor screening and in vivo drug treatment were employed to clarify the difference in drug response between the different subsets of LICs. In this study, we observed an aged Setd2 −/− mouse developing AML with co‐mutation of Nras G12S and Braf K520E. Further investigation identified two types of LICs residing in the c‐Kit+B220+Mac‐1− and c‐Kit+B220+Mac‐1+ subsets, respectively. In vivo transplantation assay disclosed the heterogeneity in differentiation between the coexistent LICs. Besides, an intrinsic doxorubicin‐resistant transcriptional signature was uncovered in c‐Kit+B220+Mac‐1+ cells. Indeed, doxorubicin plus cytarabine (DA), the standard chemotherapeutic regimen used in AML treatment, could specifically kill c‐Kit+B220+Mac‐1− cells, but it hardly affected c‐Kit+B220+Mac‐1+ cells. Transcriptome analysis unveiled a higher activation of RAS downstream signaling pathways in c‐Kit+B220+Mac‐1+ cells than in c‐Kit+B220+Mac‐1− cells. Combined treatment with DA and RAS pathway inhibitors killed both c‐Kit+B220+Mac‐1− and c‐Kit+B220+Mac‐1+ cells and attenuated disease progression. This study identified two cell subsets enriched for LICs in murine Setd2−/− ‐AML and disclosed the transcriptional and functional heterogeneity of LICs, revealing that the coexistence of different types of LICs in this model brings about diverse drug response. Transcriptional and functional heterogeneity of LICs in murine Setd2−/−‐AML was explored. Coexistence of two different types of LICs in this model brings about diverse chemo‐response. Combined treatment with DA and RAS pathway inhibitors could kill both kinds of LICs and attenuate the disease progression. These findings may advance our understanding of the intra‐heterogeneity of LICs and help us to develop more efficient treatments.
登录
查看更多内容
影响因子：
64.5
作者：
Heidorn SJ;Milagre C;Whittaker S;Nourry A;Niculescu-Duvas I;Dhomen N;Hussain J;Reis-Filho JS;Springer CJ;Pritchard C;Marais R
通讯作者：
Marais R
影响因子：
64.5
作者：
Baryawno, Ninib;Przybylski, Dariusz;Scadden, David T.
通讯作者：
Scadden, David T.
影响因子：
64.5
作者：
Burgess MR;Hwang E;Mroue R;Bielski CM;Wandler AM;Huang BJ;Firestone AJ;Young A;Lacap JA;Crocker L;Asthana S;Davis EM;Xu J;Akagi K;Le Beau MM;Li Q;Haley B;Stokoe D;Sampath D;Taylor BS;Evangelista M;Shannon K
通讯作者：
Shannon K
影响因子：
64.8
作者：
Busch, Katrin;Klapproth, Kay;Rodewald, Hans-Reimer
通讯作者：
Rodewald, Hans-Reimer
影响因子：
82.9
作者：
Eppert, Kolja;Takenaka, Katsuto;Dick, John E.
通讯作者：
Dick, John E.