DataSheet_1_Anti-HPV16 oncoproteins siRNA therapy for cervical cancer using a novel transdermal peptide PKU12.docx

In this study, an innovative transdermal peptide, #PKU12, was developed based on transdermal peptide TD-1, and the anti-tumor effect of PKU12-based siRNA against HPV was investigated in vivo. Furthermore, transcriptome differences between PKU12 + siRNA treatment and control groups were compared to assess treatment effects. The top five upregulated and downregulated genes identified by RNA sequencing were further subjected to survival analysis. The present study, for the first time, showed that this novel peptide could enhance the transdermal delivery of the siRNA targeting HPV16 L1, E6, and E7. PKU12-based siRNA delivery significantly repressed the mRNA expression levels of HPV16 L1, E6, and E7 in the SiHa xenograft tumors and attenuated tumor growth as well. The RNA-sequencing results showed that a total of 586 DEGs were detected in the PKU12 + siRNA-treated tumor tissues compared to the control tumor tissues. The GSEA analysis revealed that DEGs were inversely associated with the HIF-1 signaling pathway, the TNF signaling pathway, the AGE-RAGE signaling pathway, the NF-kappa B signaling pathway, ferroptosis, the IL-17 signaling pathway, ovarian steroidogenesis, and rheumatoid arthritis. Further functional enrichment analysis revealed that DEGs were significantly enriched in several key pathways, including cytokine–cytokine receptor interaction, the TNF signaling pathway, and the IL-17 signaling pathway. High expression of MYH1, MYH4, FGG, DEPP1, and ZBTB16 was associated with shorter overall survival of patients with cervical cancer; high expression of SULT1E1, RAB3C, CXCR3, and PROX2 was associated with longer overall survival of patients with cervical cancer. In conclusion, the transdermal peptide PKU12 is potentially a good candidate for a siRNA delivery vehicle for the treatment of cervical cancer.

本研究以透皮肽TD-1为基础，开发了一种新型透皮肽#PKU12，并在体内探究了基于PKU12的小干扰RNA（siRNA）靶向人乳头瘤病毒（HPV）的抗肿瘤效果。此外，本研究比较了PKU12+siRNA处理组与对照组的转录组差异，以评估治疗效应。通过RNA测序（RNA-seq）筛选得到的上调与下调排名前五的基因，进一步开展了生存分析。本研究首次证实，该新型肽可增强靶向HPV16 L1、E6及E7基因的siRNA的透皮递送效率。基于PKU12的siRNA递送系统可显著抑制SiHa异种移植瘤中HPV16 L1、E6及E7的mRNA表达水平，同时延缓肿瘤生长。RNA测序结果显示，与对照组肿瘤组织相比，PKU12+siRNA处理的肿瘤组织中共检测到586个差异表达基因（DEGs）。基因集富集分析（GSEA）结果表明，DEGs与缺氧诱导因子-1（HIF-1）信号通路、肿瘤坏死因子（TNF）信号通路、晚期糖基化终末产物-晚期糖基化终末产物受体（AGE-RAGE）信号通路、核因子-κB（NF-κB）信号通路、铁死亡、白细胞介素-17（IL-17）信号通路、卵巢类固醇生成以及类风湿关节炎呈负相关。进一步的功能富集分析显示，DEGs显著富集于多个关键通路，包括细胞因子-细胞因子受体相互作用、TNF信号通路以及IL-17信号通路。MYH1、MYH4、FGG、DEPP1及ZBTB16的高表达与宫颈癌患者更短的总生存期相关；而SULT1E1、RAB3C、CXCR3及PROX2的高表达则与宫颈癌患者更长的总生存期相关。综上，透皮肽PKU12有望成为治疗宫颈癌的siRNA递送载体的优质候选者。