Site-Specific HPV18 Integration Facilitates Cervical Carcinogenesis through metabolic reprogramming-Induced dysfunction of the SpHK1/S1P/S1PR1 pathway-part2

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. We employed CRISPR/Cas9 gene editing combined with homologous end repair for the establishment of stable 8q24 site-specific HPV18 gene knock-in cell lines using HaCaT as the parent. The HPV-KI cells were treated with 2-DG, FSG67, W146 and JTE013 to explore the metabolic proramming in HPV-KI/HaCaT