同源重组 (HR) 介导的 DNA 修复是维持基因组稳定性的先决条件。聚（ADP-核糖）聚合酶抑制剂（PARPis）选择性消除 HR 缺陷（HRD）的癌细胞。迄今为止，HR 相关基因测序和分析基因组不稳定性已被用作 PARPi 治疗的临床预测。然而，这些基因检测不能反映HR状态的动态变化。在这里，我们开发了一种基于病毒和活动的功能测定，可以直接量化实时 HR 活动。我们的功能检测不是关注一些 HR 相关基因，而是检测终点 HR 活性，并建立一个活性阈值来识别跨癌症类型的 HRD，并通过 PARPi 敏感性和 BRCA 状态进行验证。值得注意的是，这种基于荧光的检测可应用于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癌细胞对 PARPis 的反应。版权所有 © 2023 作者。由爱思唯尔公司出版。保留所有权利。

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.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.