Roering Pia, Siddiqui Arafat, Heuser Vanina D, Potdar Swapnil, Mikkonen Piia, Oikkonen Jaana, Li Yilin, Pikkusaari Sanna, Wennerberg Krister, Hynninen Johanna, Grenman Seija, Huhtinen Kaisa, Auranen Annika, Carpén Olli, Kaipio Katja
Institute of Biomedicine and Finnish Cancer Center (FICAN) West Cancer Centre, University of Turku and Turku University Hospital, Turku, Finland.
High Throughput Biomedicine Unit, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Helsinki, Finland.
Front Oncol. 2022 Aug 23;12:954430. doi: 10.3389/fonc.2022.954430. eCollection 2022.
A major challenge in the treatment of platinum-resistant high-grade serous ovarian cancer (HGSOC) is lack of effective therapies. Much of ongoing research on drug candidates relies on HGSOC cell lines that are poorly documented. The goal of this study was to screen for effective, state-of-the-art drug candidates using primary HGSOC cells. In addition, our aim was to dissect the inhibitory activities of Wee1 inhibitor adavosertib on primary and conventional HGSOC cell lines.
A comprehensive drug sensitivity and resistance testing (DSRT) on 306 drug compounds was performed on three patient-derived genetically unique HGSOC cell lines and two commonly used ovarian cancer cell lines. The effect of adavosertib on the cell lines was tested in several assays, including cell-cycle analysis, apoptosis induction, proliferation, wound healing, DNA damage, and effect on nuclear integrity.
Several compounds exerted cytotoxic activity toward all cell lines, when tested in both adherent and spheroid conditions. In further cytotoxicity tests, adavosertib exerted the most consistent cytotoxic activity. Adavosertib affected cell-cycle control in patient-derived and conventional HGSOC cells, inducing G2/M accumulation and reducing cyclin B1 levels. It induced apoptosis and inhibited proliferation and migration in all cell lines. Furthermore, the DNA damage marker γH2AX and the number of abnormal cell nuclei were clearly increased following adavosertib treatment. Based on the homologous recombination (HR) signature and functional HR assays of the cell lines, the effects of adavosertib were independent of the cells' HR status.
Our study indicates that Wee1 inhibitor adavosertib affects several critical functions related to proliferation, cell cycle and division, apoptosis, and invasion. Importantly, the effects are consistent in all tested cell lines, including primary HGSOC cells, and independent of the HR status of the cells. Wee1 inhibition may thus provide treatment opportunities especially for patients, whose cancer has acquired resistance to platinum-based chemotherapy or PARP inhibitors.
铂耐药高级别浆液性卵巢癌（HGSOC）治疗中的一个主要挑战是缺乏有效的治疗方法。目前许多关于候选药物的研究都依赖于记录不完善的HGSOC细胞系。本研究的目的是使用原发性HGSOC细胞筛选有效的、先进的候选药物。此外，我们的目的是剖析Wee1抑制剂阿伐替尼对原发性和传统HGSOC细胞系的抑制活性。
对三种源自患者的基因独特的HGSOC细胞系和两种常用的卵巢癌细胞系进行了306种药物化合物的全面药物敏感性和耐药性测试（DSRT）。通过几种试验测试了阿伐替尼对细胞系的作用，包括细胞周期分析、凋亡诱导、增殖、伤口愈合、DNA损伤以及对核完整性的影响。
在贴壁和球体条件下进行测试时，几种化合物对所有细胞系均表现出细胞毒性活性。在进一步的细胞毒性测试中，阿伐替尼表现出最一致的细胞毒性活性。阿伐替尼影响源自患者的和传统的HGSOC细胞中的细胞周期控制，诱导G2/M期积累并降低细胞周期蛋白B1水平。它在所有细胞系中诱导凋亡并抑制增殖和迁移。此外，阿伐替尼治疗后，DNA损伤标志物γH2AX和异常细胞核数量明显增加。基于细胞系的同源重组（HR）特征和功能性HR分析，阿伐替尼的作用与细胞的HR状态无关。
我们的研究表明，Wee1抑制剂阿伐替尼影响与增殖、细胞周期和分裂、凋亡及侵袭相关的几个关键功能。重要的是，这些作用在所有测试的细胞系中都是一致的，包括原发性HGSOC细胞，并且与细胞的HR状态无关。因此，抑制Wee1可能为尤其是那些癌症已对铂类化疗或PARP抑制剂产生耐药性的患者提供治疗机会。
