Characterization of Intrinsic Radiation Sensitivity in a Diverse Panel of Normal, Cancerous and CRISPR-Modified Cell Lines.
Characterization of Intrinsic Radiation Sensitivity in a Diverse Panel of Normal, Cancerous and CRISPR-Modified Cell Lines.
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DOI：
10.3390/ijms24097861
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发表时间：
2023-04-26
影响因子：
5.6
通讯作者：
McMahon, Stephen J. J.
中科院分区：
文献类型：
--
作者：
Liberal, Francisco D. C. Guerra;McMahon, Stephen J. J.
固有的放射敏感性是辐射反应的一个主要决定因素。尽管有大量的放射生物学数据可用，但不同研究之间的差异使生产高质量的辐射敏感性生物标志物或预测模型变得非常困难。在这里，我们描述了一组27个人类细胞系的特征，包括来自肺癌、前列腺癌和正常组织的细胞系。此外，我们使用CRISPR-Cas9生成了一组具有已知DNA修复缺陷的线。通过测量一系列生物学特征来表征这些细胞，包括X射线照射后DNA双链断裂(DSB)的诱导和修复、细胞周期分布、倍体和克隆存活。这些结果为模型开发提供了一个稳健的数据集，而不存在实验间的变异性。此外，我们使用这些结果来探索辐射敏感性的潜在决定因素之间的相关性。细胞系的本征放射敏感性差异很大，细胞系的平均失活剂量(MID)为1.3GY~3.4GY，LIG4−/−细胞的MID低至0.65GY。其他参数也有类似的显著差异，包括基线DNA损伤、细胞培养效率和倍性。在CRISPR修饰的细胞系中，残留的双链断裂能很好地预测细胞的存活(R2=0.78，p=0.009)，诱导的双链断裂水平也是很好的预测指标(R2=0.61，p=0.01)。然而，在正常细胞和癌细胞中，没有一个测量的参数与MID(R2=0.45)有很强的相关性，唯一具有统计学意义的相关指标是细胞培养效率(R2=0.31，P=0.005)和S期细胞百分比(R2=0.37，P=0.005)。虽然这些数据为放射生物学反应的建模提供了有价值的数据集，但CRISPR修饰的亚组和其他亚组之间残留DSB的预测能力的差异表明，其他途径的遗传变化，如增殖和代谢，可能对细胞辐射反应产生更大的影响。这些途径在反应模型中经常被忽略，应该在未来加以考虑。
Intrinsic radiosensitivity is a major determinant of radiation response. Despite the extensive amount of radiobiological data available, variability among different studies makes it very difficult to produce high-quality radiosensitivity biomarkers or predictive models. Here, we characterize a panel of 27 human cell lines, including those derived from lung cancer, prostate cancer, and normal tissues. In addition, we used CRISPR-Cas9 to generate a panel of lines with known DNA repair defects. These cells were characterised by measuring a range of biological features, including the induction and repair of DNA double-strand breaks (DSBs), cell cycle distribution, ploidy, and clonogenic survival following X-ray irradiation. These results offer a robust dataset without inter-experimental variabilities for model development. In addition, we used these results to explore correlations between potential determinants of radiosensitivity. There was a wide variation in the intrinsic radiosensitivity of cell lines, with cell line Mean Inactivation Doses (MID) ranging from 1.3 to 3.4 Gy for cell lines, and as low as 0.65 Gy in Lig4−/− cells. Similar substantial variability was seen in the other parameters, including baseline DNA damage, plating efficiency, and ploidy. In the CRISPR-modified cell lines, residual DSBs were good predictors of cell survival (R2 = 0.78, p = 0.009), as were induced levels of DSBs (R2 = 0.61, p = 0.01). However, amongst the normal and cancerous cells, none of the measured parameters correlated strongly with MID (R2 < 0.45), and the only metrics with statistically significant associations are plating efficiency (R2 = 0.31, p = 0.01) and percentage of cell in S phase (R2 = 0.37, p = 0.005). While these data provide a valuable dataset for the modelling of radiobiological responses, the differences in the predictive power of residual DSBs between CRISPR-modified and other subgroups suggest that genetic alterations in other pathways, such as proliferation and metabolism, may have a greater impact on cellular radiation response. These pathways are often neglected in response modelling and should be considered in the future.
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影响因子：
4.7
作者：
McMahon SJ;Prise KM
通讯作者：
Prise KM
影响因子：
3.8
作者：
Banáth JP;Klokov D;MacPhail SH;Banuelos CA;Olive PL
通讯作者：
Olive PL
内在放射敏性作为对子宫颈癌放射疗法的患者反应的预后因素的独立性。
影响因子：
8.8
作者：
West, C M;Davidson, S E;Roberts, S A;Hunter, R D
通讯作者：
Hunter, R D
影响因子：
3.7
作者：
Hall JS;Iype R;Senra J;Taylor J;Armenoult L;Oguejiofor K;Li Y;Stratford I;Stern PL;O'Connor MJ;Miller CJ;West CM
通讯作者：
West CM
影响因子：
2.7
作者：
Heaven CJ;Wanstall HC;Henthorn NT;Warmenhoven JW;Ingram SP;Chadwick AL;Santina E;Honeychurch J;Schmidt CK;Kirkby KJ;Kirkby NF;Burnet NG;Merchant MJ
通讯作者：
Merchant MJ