Effect of the evolutionary selected Andean Aymara NFKB1 polymorphisms on inflammatory and HIF-dependent gene expression in endometrial and ovarian cancer

JS,SS-corresponding authors; SJK, AP-equal contributionGynecological cancers cause ~132/1 million deaths in women /year worldwide (PMID: 39148469). Endometrial cancer (EC) is the most common gynecological cancer, while ovarian cancer (OC) is the most lethal, with a 5-year US mortality ~51% overall and less than 30% in advanced stage of disease. NF-κB signaling contributes to progression, chemoresistance, and relapse in both EC and OC. The evolutionary selected Aymara NFKB1 variant (rs230511; CC, CT, TT genotype), existing in ~30% Europeans, Asians, and Hispanics, correlates with expression of inflammatory and hypoxia-inducible factor (HIF)-regulated genes (PMID:39971917). Because inflammation and hypoxia promote tumor growth, invasion, metastasis, and resistance to treatment, this variant may influence EC/OC biology and clinical outcomes. We investigated role rs230511 genotypes in inflammatory and HIF-dependent gene expression in EC and OC.Genomic DNA from blood mononuclear cells from EC (n=103), OC (n=108) and benign controls (n=86) was genotyped for rs230511. Tumors samples from treatment naïve patients (25 OC, and 20 EC) and normal tissues from (ovary n=6; endometrium n=9) were collected. RNA was extracted and gene expression analysis of HIF-targeted genes ( VEGFA, SLC2A1, LDHA) and inflammatory genes (IL6, P2RY2, CCR7, CXCL8, IL1B, NFKB1 , TNF ), with GAPDH as the housekeeping gene evaluated . No differences in allele frequencies or genotype distributions of rs230511 were observed among EC, OC, and benign groups, indicating that this NFKB1 variant does not predispose to OC/EC. In EC tumors, NFKB1 mRNA were significantly lower than in normal endometrium (p=0.004) NFKB1 expression in EC positively correlated with the expression of HIF-targeted genes ( VEGFA , SLC2A1 ) and inflammatory genes ( IL6, P2RY2, TNF ). Patients carrying the CT genotype had lower expression of IL6 and P2RY2 than the CC , suggesting less aggressive tumors.; the expression of other inflammatory genes ( CCR7, CXCL8, IL1B, TNF) also trended lower but were not significant. The CT genotype was associated with lower white blood cell counts than CC , consistent with reduced inflammation. In OC, NFKB1 mRNA was lower than in normal ovary. OC patients with the CT and TT genotype had lower NFKB1 mRNA than CC carriers. In OC inflammatory genes ( CCR7, CXCL8, IL1B , TNF ) were highly expressed but did not correlate with NFKB1 mRNA levels and genotypes. HIF-target genes positively correlated with inflammatory gene levels, suggesting HIF, not NFKB1, drives inflammation in OC. NFKB1 genotype-dependent variation and its downstream pathways appear to influence EC but not OC, potentially contributing to differences in tumor aggressiveness and prognosis. The effect of the NFKB1 genotype is undergoing analysis of overall survival, tumor aggressiveness and treatment response.