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

<p><span>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&mdash;PD-1 monoclonal antibody (PD-1) therapy and whole cancer cell vaccine (WCV) therapy&mdash;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&rarr;IL18R1&rarr;p-JNK&rarr;COL1A1, promoting primordial follicle overactivation through CXCL10&rarr;IL18R1&rarr;p-AKT, and increasing ovarian inflammation through CXCL10&rarr;IL18R1&rarr;p-P65. Finally, we rescued the decreased ovarian function in the M group by blocking the CXCL10&rarr;IL18R1 pathway with CXCL10 antibody or a CXCL10&ndash;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.</span></p> <p><strong><span>Supplementary dataset 1</span></strong></p> <p><span>Related to Fig. 3A and 3B. This file includes two sheets. &ldquo;All_samples-fpkm&rdquo; includes FPKM values of all identified genes in ovaries from CTR, M, VM, and PM group (three repeats per group); &ldquo;heat map&rdquo; includes all average log2 values (from three repeats per group) of 435 DEGs (</span><span>differentially expressed genes</span><span>) </span><span>at the threshold of |log2(M/CTR)| &ge; 1.</span></p> <p><strong><span>Supplementary dataset 2</span></strong></p> <p><span>Related to Fig. 3C. This file includes 33 sheets. &ldquo;summary&rdquo; includes all conc. values (pg/ml) of 31 plasma DECs (</span><span>differentially expressed cytokines</span><span>); &ldquo;Curves&rdquo; 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).</span></p> <p><strong><span>Supplementary dataset 3</span></strong></p> <p><span>Related to Fig. 4A. This file includes two sheets. &ldquo;peptides&rdquo; includes all related values of all identified peptides; &ldquo;Protein Groups&rdquo; includes the related info. Of all proteins corresponding to the identified peptides</span></p>