Variation in RARG increases susceptibility to doxorubicin-induced cardiotoxicity in patient specific induced pluripotent stem cell-derived cardiomyocytes
Variation in RARG increases susceptibility to doxorubicin-induced cardiotoxicity in patient specific induced pluripotent stem cell-derived cardiomyocytes
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DOI：
10.1038/s41598-020-65979-x
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发表时间：
2020-06-25
影响因子：
4.6
通讯作者：
Brunham, Liam R.
中科院分区：
文献类型：
--
作者：
Christidi, Effimia;Huang, Haojun;Brunham, Liam R.
阿霉素是一种有效的抗癌药物，用于治疗各种癌症类型。然而，它的使用受到阿霉素诱导的心脏毒性（DIC）的限制。在一项全基因组关联研究中，RARG基因（S427 L; rs 2229774）中的错义变体与DIC的易感性有关。本研究的目的是调查这种RARG变异体在DIC中的功能作用。我们使用来自用阿霉素治疗的患者的诱导多能干细胞衍生的心肌细胞（iPSC-CM）。在细胞活力和光学图谱实验中，与来自多柔比星治疗的未发生DIC的个体（对照）的iPSC-CM相比，来自经历DIC的个体（病例）的iPSC-CM显示出对多柔比星显著更高的敏感性。使用CRISPR/Cas9，我们产生了仅在RARG基因座上不同的等基因细胞系。RARG-S427 L对野生型的遗传校正导致减少阿霉素诱导的双链DNA断裂、活性氧产生和细胞死亡。相反，RARG-S427 L的引入增加了对多柔比星的易感性。最后，RARG基因的遗传破坏导致保护免受多柔比星治疗引起的细胞死亡。我们的研究结果表明，RARG-S427 L的存在增加了对DIC的敏感性，建立了该变体在DIC中的直接因果作用。
Doxorubicin is a potent anticancer drug used to treat a variety of cancer types. However, its use is limited by doxorubicin-induced cardiotoxicity (DIC). A missense variant in the RARG gene (S427L; rs2229774) has been implicated in susceptibility to DIC in a genome wide association study. The goal of this study was to investigate the functional role of this RARG variant in DIC. We used induced pluripotent stem cell derived cardiomyocytes (iPSC-CMs) from patients treated with doxorubicin. iPSC-CMs from individuals who experienced DIC (cases) showed significantly greater sensitivity to doxorubicin compared to iPSC-CMs from doxorubicin-treated individuals who did not develop DIC (controls) in cell viability and optical mapping experiments. Using CRISPR/Cas9, we generated isogenic cell lines that differed only at the RARG locus. Genetic correction of RARG-S427L to wild type resulted in reduced doxorubicin-induced double stranded DNA breaks, reactive oxygen species production, and cell death. Conversely, introduction of RARG-S427L increased susceptibility to doxorubicin. Finally, genetic disruption of the RARG gene resulted in protection from cell death due to doxorubicin treatment. Our findings suggest that the presence of RARG-S427L increases sensitivity to DIC, establishing a direct, causal role for this variant in DIC.