Targeting novel m6A reader KHSRP impairs pancreatic ductal adenocarcinoma progression via attenuating FAK signaling

We report KH-type splicing regulatory protein (KHSRP) as a novel m6A reader with oncogenic functions in pancreatic ductal adenocarcinoma (PDAC). KHSRP recognizes and stabilizes its target mRNAs (e.g., MET, ITGAV and ITGB1) in an m6A-dependent manner, therefore activating downstream FAK signaling and promoting PDAC progression. Targeting KHSRP shows promising anti-tumoral effects in vitro and in vivo, indicating that KHSRP may serve as a therapeutic target for PDAC. We performed m6A individual-nucleotide resolution cross-linking and immunoprecipitation sequencing (miCLIP-seq) and photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation sequencing (PAR-CLIP-seq) for KHSRP to investigate whether KHSRP possesses the characteristics of m6A-binding protein. To explore the underlying mechanism of KHSRP-mediated tumor progression, we conducted RNA sequencing (RNA-seq). We further performed mRNA stability analysis to examine the effect of KHSRP on mRNA lifetime.

本研究报道KH型剪接调控蛋白（KH-type splicing regulatory protein, KHSRP）是一种新型的m6A读取蛋白（m6A reader），在胰腺导管腺癌（pancreatic ductal adenocarcinoma, PDAC）中发挥致癌功能。KHSRP以m6A依赖的方式识别并稳定其靶mRNA（如MET、ITGAV及ITGB1），进而激活下游FAK信号通路，促进PDAC进展。靶向KHSRP在体外与体内均展现出良好的抗肿瘤效应，提示KHSRP可作为PDAC的潜在治疗靶点。为探究KHSRP是否具备m6A结合蛋白的特性，我们针对KHSRP开展了m6A单核苷酸分辨率交联免疫沉淀测序（m6A individual-nucleotide resolution cross-linking and immunoprecipitation sequencing, miCLIP-seq）与光活化核糖核苷增强交联免疫沉淀测序（photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation sequencing, PAR-CLIP-seq）。为解析KHSRP介导的肿瘤进展的潜在分子机制，我们开展了RNA测序（RNA-seq）。我们进一步开展了mRNA稳定性分析，以验证KHSRP对mRNA半衰期的调控作用。