Whole Cancer Cell Vaccine Protects Ovarian Functions in Tumor-bearing Mice through Downregulating CXCL10

Young female patients with cancer are likely to become sub-fertile or infertile even if they ultimately overcome cancer through various therapies. Cancer immunotherapy has recently emerged as a promising novel therapy against cancers with high malignancy and lethality, but it is unclear whether cancer immunotherapy affects female fertility. This study employed MCA205 cell-allotransplanted B6 mice as a model to investigate whether two popular immunotherapies—PD-1 monoclonal antibody (PD-1) therapy and whole cancer cell vaccine (WCV) therapy—affect ovarian function. MCA205 allotransplanted (M) mice exhibited decreased follicle numbers at each stage, decreased proliferation, increased apoptosis, and a decreased oocyte maturation rate. WCV treatment significantly reversed these abnormalities, whereas PD-1 did not. RNA sequencing of the ovaries revealed that multiple differentially expressed genes (DEGs) were involved in inflammation pathways. Furthermore, cytokine microarray characterized CXCL10 with both biggest increment in M group and best rescue in WCV group. Next, CXCL10 antibody Immunoprecipitation in ovarian lysate and LC-MS baited the only receptor IL18R1. Furthermore, we found that CXCL10 impaired ovarian function through three pathways: inducing ovarian fibrosis through CXCL10→IL18R1→p-JNK→COL1A1, promoting primordial follicle overactivation through CXCL10→IL18R1→p-AKT, and increasing ovarian inflammation through CXCL10→IL18R1→p-P65. Finally, we rescued the decreased ovarian function in the M group by blocking the CXCL10→IL18R1 pathway with CXCL10 antibody or a CXCL10–IL18R1 interface peptide, CIBB. This study provides mechanical evidence and translational strategies for WCVs to achieve the dual functions of suppressing tumor progression while protecting ovarian function.

Supplementary dataset 1

Related to Fig. 3A and 3B. This file includes two sheets. “All_samples-fpkm” includes FPKM values of all identified genes in ovaries from CTR, M, VM, and PM group (three repeats per group); “heat map” includes all average log2 values (from three repeats per group) of 435 DEGs (differentially expressed genes) at the threshold of |log2(M/CTR)| ≥ 1.

Supplementary dataset 2

Related to Fig. 3C. This file includes 33 sheets. “summary” includes all conc. values (pg/ml) of 31 plasma DECs (differentially expressed cytokines); “Curves” includes standard curves for all 31 cytokines; each of the other 31 sheets include the detailed assays for each plasma cytokine for CTR, M, VM, and PM group (five repeats per group).

Supplementary dataset 3

Related to Fig. 4A. This file includes two sheets. “peptides” includes all related values of all identified peptides; “Protein Groups” includes the related info. Of all proteins corresponding to the identified peptides