Genome-wide CRISPR/Cas9 screening identifies a targetable MEST-PURA interaction in cancer metastasis

作者：Wen Xu, Long Liao, Wei Dai, Can‐Can Zheng, Xiangpeng Tan, Yan He, Qihua Zhang, Zhi‐Hao Huang, Wen‐You Chen, Yanru Qin, Kui-Sheng Chen, Ming‐Liang He, Simon Law, Maria Li Lung, Qing‐Yu He, Bin Li · 发表于：EBioMedicine · 年份：2023 · DOI：10.1016/j.ebiom.2023.104587 · 被引用次数：26 · 研究领域：Epigenetics and DNA Methylation、Cancer Cells and Metastasis、MicroRNA in disease regulation

BACKGROUND: Metastasis is one of the most lethal hallmarks of esophageal squamous cell carcinoma (ESCC), yet the mechanisms remain unclear due to a lack of reliable experimental models and systematic identification of key drivers. There is urgent need to develop useful therapies for this lethal disease. METHODS: A genome-wide CRISPR/Cas9 screening, in combination with gene profiling of highly invasive and metastatic ESCC sublines, as well as PDX models, was performed to identify key regulators of cancer metastasis. The Gain- and loss-of-function experiments were taken to examine gene function. Protein interactome, RNA-seq, and whole genome methylation sequencing were used to investigate gene regulation and molecular mechanisms. Clinical significance was analyzed in tumor tissue microarray and TCGA databases. Homology modeling, modified ELISA, surface plasmon resonance and functional assays were performed to identify lead compound which targets MEST to suppress cancer metastasis. FINDINGS: High MEST expression was associated with poor patient survival and promoted cancer invasion and metastasis in ESCC. Mechanistically, MEST activates SRCIN1/RASAL1-ERK-snail signaling by interacting with PURA. miR-449a was identified as a direct regulator of MEST, and hypermethylation of its promoter led to MEST upregulation, whereas systemically delivered miR-449a mimic could suppress tumor metastasis without overt toxicity. Furthermore, molecular docking and computational screening in a smal...