Institute of Biochemical Sciences, National Taiwan University, Taipei, Taiwan.
Department of Obstetrics & Gynecology, National Taiwan University Hospital, Taipei, Taiwan.
Cell Rep Med. 2023 Nov 21;4(11):101247. doi: 10.1016/j.xcrm.2023.101247. Epub 2023 Oct 19.
Homologous recombination (HR)-mediated DNA repair is a prerequisite for maintaining genome stability. Cancer cells displaying HR deficiency (HRD) are selectively eliminated by poly(ADP-ribose) polymerase inhibitors (PARPis). To date, sequencing of HR-associated genes and analyzing genome instability have been used as clinical predictions for PARPi therapy. However, these genetic tests cannot reflect dynamic changes in the HR status. Here, we have developed a virus- and activity-based functional assay to quantify real-time HR activity directly. Instead of focusing on a few HR-associated genes, our functional assay detects endpoint HR activity and establishes an activity threshold for identifying HRD across cancer types, validated by PARPi sensitivity and BRCA status. Notably, this fluorescence-based assay can be applied to primary ovarian cancer cells from patients to reflect their level of HRD, which is associated with survival benefits. Thus, our work provides a functional test to predict the response of primary cancer cells to PARPis.
同源重组 (HR)-介导的 DNA 修复是维持基因组稳定性的前提。显示 HR 缺陷 (HRD) 的癌细胞被聚 (ADP-核糖) 聚合酶抑制剂 (PARPi) 选择性消除。迄今为止，HR 相关基因的测序和基因组不稳定性分析已被用作 PARPi 治疗的临床预测。然而，这些遗传测试不能反映 HR 状态的动态变化。在这里，我们开发了一种基于病毒和活性的功能测定法来直接定量实时 HR 活性。与专注于少数 HR 相关基因的方法不同，我们的功能测定法检测终点 HR 活性，并建立了一种活性阈值，用于在癌症类型中识别 HRD，该阈值通过 PARPi 敏感性和 BRCA 状态得到验证。值得注意的是，这种基于荧光的测定法可应用于来自患者的原发性卵巢癌细胞，以反映其 HRD 水平，这与生存获益相关。因此，我们的工作提供了一种功能测试来预测原发性癌细胞对 PARPis 的反应。
