Cooperative adaptation to therapy (CAT) confers resistance in heterogeneous non-small cell lung cancer
Cooperative adaptation to therapy (CAT) confers resistance in heterogeneous non-small cell lung cancer
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DOI：
10.1371/journal.pcbi.1007278
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发表时间：
2019-08-01
影响因子：
4.3
通讯作者：
Goldman, Aaron
中科院分区：
文献类型：
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作者：
Craig, Morgan;Kaveh, Kamran;Goldman, Aaron
了解内在和获得性耐药对于克服癌症化疗失败至关重要。虽然众所周知，肿瘤内、亚克隆遗传和表型的异质性对耐药性有显著贡献，但肿瘤亚克隆如何相互作用以承受治疗压力尚不完全清楚。在这里，我们报告了在异质肿瘤中一种以前未被认识的行为：合作适应治疗(CAT)，即当癌细胞暴露在癌症治疗中时，在邻近的癌细胞中诱导出共同耐药的表型。使用CRISPR/Cas9工具包，我们通过诱导Dicer1的突变来设计表型多样化的非小细胞肺癌(NSCLC)细胞。Dicer1是一种参与小分子非编码RNA发生的III型细胞质内切核酸酶。我们使用无底物纳米培养系统，在无刺激或药物压力条件下，单独或共培养时监测了荧光标记突变细胞和/或野生型细胞的三维生长动力学。通过将数学建模与流式细胞术相结合，我们使用了种内和种间竞争的数学模型来表征单一和共培养的生长模式。利用流式细胞仪数据，我们估计了模型的参数，以揭示WT和突变体在共培养中的组合允许在以前对药物敏感的细胞中有益的生长，尽管药物压力通过诱导细胞状态转换来描述，该变化由合作博弈论的适应值变化描述。最后，我们使用了一个体外人类肿瘤模型，该模型通过药物敏感性分析来预测临床反应，并在个体NSCLC患者样本中确定细胞和形态的异质性与多种临床批准和标签外药物的预后失败相关。总之，这些发现提出了一个新的抗药性悖论，涉及肿瘤细胞之间的非遗传合作来阻止药物压力，这表明仅对可药物靶点(即突变)的描述可能不足以分配有效的治疗。
Understanding intrinsic and acquired resistance is crucial to overcoming cancer chemotherapy failure. While it is well-established that intratumor, subclonal genetic and phenotypic heterogeneity significantly contribute to resistance, it is not fully understood how tumor sub-clones interact with each other to withstand therapy pressure. Here, we report a previously unrecognized behavior in heterogeneous tumors: cooperative adaptation to therapy (CAT), in which cancer cells induce co-resistant phenotypes in neighboring cancer cells when exposed to cancer therapy. Using a CRISPR/Cas9 toolkit we engineered phenotypically diverse non-small cell lung cancer (NSCLC) cells by conferring mutations in Dicer1, a type III cytoplasmic endoribonuclease involved in small non-coding RNA genesis. We monitored three-dimensional growth dynamics of fluorescently-labeled mutant and/or wild-type cells individually or in co-culture using a substrate-free NanoCulture system under unstimulated or drug pressure conditions. By integrating mathematical modeling with flow cytometry, we characterized the growth patterns of mono- and co-cultures using a mathematical model of intra- and interspecies competition. Leveraging the flow cytometry data, we estimated the model's parameters to reveal that the combination of WT and mutants in co-cultures allowed for beneficial growth in previously drug sensitive cells despite drug pressure via induction of cell state transitions described by a cooperative game theoretic change in the fitness values. Finally, we used an ex vivo human tumor model that predicts clinical response through drug sensitivity analyses and determined that cellular and morphologic heterogeneity correlates to prognostic failure of multiple clinically-approved and off-label drugs in individual NSCLC patient samples. Together, these findings present a new paradox in drug resistance implicating non-genetic cooperation among tumor cells to thwart drug pressure, suggesting that profiling for druggable targets (i.e. mutations) alone may be insufficient to assign effective therapy.