PDF(Pubmed)
Abstract：
OBJECTIVE: With the development of immunotherapy research, the role of immune checkpoint blockade (ICB) in the treatment of cervical cancer has been emphasized, but many patients still can\'t receive long-term benefits from ICB. Poly ADP ribose polymerase inhibitor (PARPi) has been proved to exert significant antitumor effects in multiple solid tumors. Whether cervical cancer patients obtain better benefits from the treatment regimen of PARPi combined with ICB remains unclear.
METHODS: The alteration of PD-L1 expression induced by niraparib in cervical cancer cells and its underlying mechanism were assessed by western blot and immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR).The regulation of PTEN by KDM5A was confirmed using Chromatin immunoprecipitation (ChIP) assay and RNA interference. Analyzing the relationship between PD-L1 and immune effector molecules through searching online databases. Therapeutic efficacy of niraparib, PD-L1 blockade or combination was assessed in syngeneic tumor model. The changes of immune cells and cytokines in vivo was detected by immunohistochemistry (IHC) and qRT-PCR.
RESULTS: We found that niraparib upregulated PD-L1 expression and potentiated the antitumor effects of PD-L1 blockade in a murine cervical cancer model. Niraparib inhibited the Pten expression by increasing the abundance of KDM5A, which expanded PD-L1 abundance through activating the PI3K-AKT-S6K1 pathway. PD-L1 was positively correlated with immune effector molecules including TNF-α, IFN-γ, granzyme A and granzyme B based on biological information analysis. Niraparib increased the infiltration of CD8+ T cells and the level of IFN-γ, granzyme B in vivo.
CONCLUSIONS: Our findings demonstrates the regulation of niraparib on local immune microenvironment of cervical cancer, and provides theoretical basis for supporting the combination of PARPi and PD-L1 blockade as a potential treatment for cervical cancer.
METHODS: The alteration of PD-L1 expression induced by niraparib in cervical cancer cells and its underlying mechanism were assessed by western blot and immunofluorescence and quantitative real-time polymerase chain reaction (qRT-PCR).The regulation of PTEN by KDM5A was confirmed using Chromatin immunoprecipitation (ChIP) assay and RNA interference. Analyzing the relationship between PD-L1 and immune effector molecules through searching online databases. Therapeutic efficacy of niraparib, PD-L1 blockade or combination was assessed in syngeneic tumor model. The changes of immune cells and cytokines in vivo was detected by immunohistochemistry (IHC) and qRT-PCR.
RESULTS: We found that niraparib upregulated PD-L1 expression and potentiated the antitumor effects of PD-L1 blockade in a murine cervical cancer model. Niraparib inhibited the Pten expression by increasing the abundance of KDM5A, which expanded PD-L1 abundance through activating the PI3K-AKT-S6K1 pathway. PD-L1 was positively correlated with immune effector molecules including TNF-α, IFN-γ, granzyme A and granzyme B based on biological information analysis. Niraparib increased the infiltration of CD8+ T cells and the level of IFN-γ, granzyme B in vivo.
CONCLUSIONS: Our findings demonstrates the regulation of niraparib on local immune microenvironment of cervical cancer, and provides theoretical basis for supporting the combination of PARPi and PD-L1 blockade as a potential treatment for cervical cancer.
摘要：
目的：随着免疫治疗研究的发展，强调了免疫检查点阻断（ICB）在宫颈癌治疗中的作用，但许多患者仍然无法从ICB中获得长期益处。聚ADP核糖聚合酶抑制剂（PARPi）已被证明在多种实体瘤中具有显着的抗肿瘤作用。宫颈癌患者是否从PARPi联合ICB的治疗方案中获得了更好的益处尚不清楚。
方法：采用westernblot、免疫荧光和实时定量聚合酶链反应(qRT-PCR)检测尼拉帕尼诱导宫颈癌细胞PD-L1表达的改变及其机制。使用染色质免疫沉淀（ChIP）测定和RNA干扰证实了KDM5A对PTEN的调节。通过搜索在线数据库分析PD-L1与免疫效应分子之间的关系。尼拉帕尼的疗效，在同基因肿瘤模型中评估PD-L1阻断或组合。免疫组织化学(IHC)和qRT-PCR检测体内免疫细胞和细胞因子的变化。
结果：我们发现尼拉帕尼在小鼠宫颈癌模型中上调PD-L1表达并增强PD-L1阻断的抗肿瘤作用。尼拉帕尼通过增加KDM5A的丰度抑制Pten的表达，通过激活PI3K-AKT-S6K1途径扩大PD-L1的丰度。PD-L1与包括TNF-α在内的免疫效应分子呈正相关，IFN-γ，基于生物信息分析的颗粒酶A和颗粒酶B。尼拉帕尼增加了CD8+T细胞的浸润和IFN-γ的水平，颗粒酶B在体内。
结论：我们的研究结果表明尼拉帕尼对宫颈癌局部免疫微环境的调节作用，为支持PARPi联合PD-L1阻断作为宫颈癌潜在治疗提供了理论依据。
方法：采用westernblot、免疫荧光和实时定量聚合酶链反应(qRT-PCR)检测尼拉帕尼诱导宫颈癌细胞PD-L1表达的改变及其机制。使用染色质免疫沉淀（ChIP）测定和RNA干扰证实了KDM5A对PTEN的调节。通过搜索在线数据库分析PD-L1与免疫效应分子之间的关系。尼拉帕尼的疗效，在同基因肿瘤模型中评估PD-L1阻断或组合。免疫组织化学(IHC)和qRT-PCR检测体内免疫细胞和细胞因子的变化。
结果：我们发现尼拉帕尼在小鼠宫颈癌模型中上调PD-L1表达并增强PD-L1阻断的抗肿瘤作用。尼拉帕尼通过增加KDM5A的丰度抑制Pten的表达，通过激活PI3K-AKT-S6K1途径扩大PD-L1的丰度。PD-L1与包括TNF-α在内的免疫效应分子呈正相关，IFN-γ，基于生物信息分析的颗粒酶A和颗粒酶B。尼拉帕尼增加了CD8+T细胞的浸润和IFN-γ的水平，颗粒酶B在体内。
结论：我们的研究结果表明尼拉帕尼对宫颈癌局部免疫微环境的调节作用，为支持PARPi联合PD-L1阻断作为宫颈癌潜在治疗提供了理论依据。
