Xiao Yinan, Wu Yu, Wang Qilong, Li Mo, Deng Chaolin, Gu Xiaoyang
Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing, 100191, China.
National Clinical Research Center for Obstetrics and Gynecology, Peking University Third Hospital), Beijing, 100191, China.
Cell Commun Signal. 2025 Jan 25;23(1):48. doi: 10.1186/s12964-025-02056-8.
Ovarian cancer (OC), particularly high-grade serous ovarian carcinoma (HGSOC), is the leading cause of mortality from gynecological malignancies worldwide. Despite the initial effectiveness of treatment, acquired resistance to poly(ADP-ribose) polymerase inhibitors (PARPis) represents a major challenge for the clinical management of HGSOC, highlighting the necessity for the development of novel therapeutic strategies. This study investigated the role of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), a pivotal regulator of glycolysis, in PARPi resistance and explored its potential as a therapeutic target to overcome PARPi resistance.
We conducted in vitro and in vivo experiments to assess the role of PFKFB3 in OC and its impact on PARPi resistance. We analyzed PFKFB3 expression and activity in primary OC tissues and cell lines using western blotting and immunohistochemistry. CRISPR-Cas9 and pharmacological inhibitors were employed to inhibit PFKFB3, and the effects on PARPi resistance, homologous recombination (HR) repair efficiency, and DNA damage were evaluated. RNA sequencing and proximity labeling were employed to identify the molecular mechanisms underlying PFKFB3-mediated resistance. The in vivo efficacy of PARPi and PFK158 combination therapy was evaluated in OC xenograft models.
PFKFB3 activity was significantly elevated in OC tissues and associated with PARPi resistance. Inhibition of PFKFB3, both genetically and pharmacologically, sensitized OC cells to PARPis, impaired HR repair and increased DNA damage. Proximity labeling revealed replication protein A3 (RPA3) as a novel PFKFB3-binding protein involved in HR repair. In vivo, the combination of PFK158 and olaparib significantly inhibited tumor growth, increased DNA damage, and induced apoptosis in OC xenografts without exacerbating adverse effects.
Our findings demonstrate that PFKFB3 is crucial for PARPi resistance in OC. Inhibiting PFKFB3 sensitizes HR-proficient OC cells to PARPis by impairing HR repair, leading to increased DNA damage and apoptosis. PFKFB3 represents a promising therapeutic target for overcoming PARPi resistance and improving outcomes in OC patients.
卵巢癌（OC），尤其是高级别浆液性卵巢癌（HGSOC），是全球妇科恶性肿瘤死亡的主要原因。尽管初始治疗有效，但对聚（ADP-核糖）聚合酶抑制剂（PARPis）产生获得性耐药是HGSOC临床管理的一大挑战，凸显了开发新型治疗策略的必要性。本研究调查了糖酵解关键调节因子6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶3（PFKFB3）在PARPi耐药中的作用，并探索其作为克服PARPi耐药的治疗靶点的潜力。
我们进行了体外和体内实验，以评估PFKFB3在OC中的作用及其对PARPi耐药的影响。我们使用蛋白质免疫印迹和免疫组织化学分析原发性OC组织和细胞系中PFKFB3的表达和活性。采用CRISPR-Cas9和药物抑制剂抑制PFKFB3，并评估其对PARPi耐药、同源重组（HR）修复效率和DNA损伤的影响。采用RNA测序和邻近标记法确定PFKFB3介导耐药的分子机制。在OC异种移植模型中评估PARPi与PFK158联合治疗的体内疗效。
PFKFB3活性在OC组织中显著升高，并与PARPi耐药相关。通过基因和药物抑制PFKFB3，使OC细胞对PARPis敏感，损害HR修复并增加DNA损伤。邻近标记显示复制蛋白A3（RPA3）是一种参与HR修复的新型PFKFB3结合蛋白。在体内，PFK158与奥拉帕利联合使用可显著抑制OC异种移植瘤的生长，增加DNA损伤并诱导凋亡，且不会加剧不良反应。
我们的研究结果表明，PFKFB3对OC中的PARPi耐药至关重要。抑制PFKFB通过损害HR修复使HR功能正常的OC细胞对PARPis敏感，导致DNA损伤增加和细胞凋亡。PFKFB3是克服PARPi耐药和改善OC患者预后的一个有前景的治疗靶点。
