Verapamil modulates NFAT2 to inhibit tumor growth and potentiates PD1ab immune checkpoint inhibitor therapy in cervical cancer treatment

AbstractPurpose Current evidence suggests a high co-prevalence of hypertension and cervical cancer. Accordingly, blood pressure control is indicated during anti-tumor drug therapy in this patient population. Over the past few years, immunotherapy has made great strides in treating different cancers. However, the role and clinical significance of verapamil as a first-line anti-hypertensive drug during immunotherapy remain poorly understood, emphasizing the need for further studies.Methods Murine cervical cancer models were employed to assess the effect of verapamil monotherapy and combination with PD1ab. Immunohistochemistry was conducted to quantify the abundance of CD8+ T cell and Ki67+ cells. Several in-vitro and in-vivo assays were used to study the effects of verapamil and explore the preliminary mechanism.Results Monotherapy with verapamil or PD1ab immune checkpoint inhibitor significantly suppressed the growth of subcutaneously grafted U14 cells in WT BABL/c mice, respectively, with increased survival time of mice. Consistent results were observed in the melanoma model. Furthermore, we substantiated that verapamil significantly impaired tumor proliferation and migration of SiHa human cervical cancer cells in vitro and in vivo. In silico analysis using TCGA data revealed that NFAT2 expression negatively correlated with patient survival. The CCK8 assay revealed that verapamil abrogated the stimulatory effect of NFAT2 after knockdown of NFAT2.Conclusions Our results suggest that verapamil inhibits tumor growth by modulating NFAT2 expression and enhancing tumor immune responses to PD1ab, which can be harnessed for cervical cancer therapy, especially for patients with comorbid hypertension. Indeed, further clinical trials are warranted to increase the robustness of our findings.Keywords: VerapamilPD1abNFAT2cervical cancer AcknowledgmentsWe thank Department of Medicine, Taizhou University for providing facilities and technical support.Author contributionsConceptualization: YQ. Methodology: YQ, BB, QX. Investigation: YQ, BB, QX. Visualization: YQ, BB, QX. Funding acquisition: YQ, BB. Project administration: YQ. Supervision: YQ, WY. Writing – original draft: YQ, WY. Writing – review & editing: YQ, LL, GM.Disclosure statementNo potential conflict of interest was reported by the author(s).Data availability statementAll data are available in the main text or the supplementary materials.Additional informationFundingThe Taizhou Municipal Science and Technology Bureau [No.21ywb112]; The Taizhou Municipal Science and Technology Bureau [No.22ywb86]; The Wenling Social Development Science and Technology Project [No.2021S00003].