Site-Specific HPV18 Integration Facilitates Cervical Carcinogenesis through metabolic reprogramming-Induced dysfunction of the SpHK1/S1P/S1PR1 pathway

The integration of high-risk human papillomavirus (HR-HPV) into specific loci of the human genome is a pivotal event in cervical carcinogenesis. However, how HPV integration contributes to malignant transformation remains unclear. Here, we utilized the CRISPR/Cas9 system established an 8q24 site-specific HPV18 gene knock-in cell model. We discovered that HPV18 knock-in resulted in a global alteration of the genome's topologically associating domain (TAD) structure and up-regulation of cancer-related genes in HaCaT cells, leading to malignant transformation. Furthermore, the IL-17 signaling pathway and S100A8/A9 genes were found to be significantly up-regulated in the HPV18 knock-in cell line, as well as K14-HPV transgenic mice and cervical cancer clinical database. Metabolic reprogramming of the HPV18 knock-in cell line, particularly the up-regulation of glycolysis, facilitated glycerolipid synthesis and Sphingosine-1-phospate(S1P) secretion. The increased secretion of S1P and activation of the S1P receptor 1(S1PR1) signaling pathway induced the expression of S100A8/A9 and cytokines. Inhibition of the S1P/S1PR1 signaling pathway down-regulated the expression of S100A8/A9 and suppressed the growth of HPV18 knock-in cell line and cervical cancer patient derived Xenograft. These findings provide novel insights into the molecular mechanisms underlying cervical carcinogenesis and identify potential therapeutic targets for its treatment.