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Esophageal squamous cell carcinoma (ESCC) is a highly aggressive cancer with poor survival, yet the key kinases driving ESCC metastasis and their biological function have not been fully discovered. Here, a kinase\u2010substrate map of metastatic ESCC is presented for the first time by conducting a phosphoproteomics analysis of 60 clinical specimens. By further consolidating data with CRISPR\/Cas9 functional screening, LIM domain kinase 1 (LIMK1) is identified as a novel kinase of \u03b2\u2010catenin. The in vitro and in vivo experiments demonstrated that LIMK1 cooperates with Cyclin\u2010dependent kinase 5 (CDK5) to promote cancer metastasis in a phosphorylation\u2010dependent manner. Mechanistically, LIMK1 and CDK5 synergistically phosphorylate \u03b2\u2010catenin at S191, enhancing its phosphorylation and interaction with Nucleoporin 93, resulting in \u03b2\u2010catenin nuclear translocation and activation of key pathways in cancer metastasis. High expression of LIMK1 and CDK5 is associated with poor prognosis of ESCC patients, and the clinical and functional significance of LIMK1\/CDK5\u2010Wnt\/\u03b2\u2010catenin axis is also verified in esophageal adenocarcinoma, gastric cancer, and lung cancer. Furthermore, the combination of LIMK1 and CDK5 inhibitors significantly suppresses metastasis in multiple models. This work highlights LIMK1 as a novel regulatory and targetable kinase of \u03b2\u2010catenin, informing the treatment of advanced cancer.<\/jats:p>","DOI":"10.1002\/advs.202503223","type":"journal-article","created":{"date-parts":[[2025,6,6]],"date-time":"2025-06-06T06:29:32Z","timestamp":1749191372000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["LIMK1 as a Novel Kinase of \u03b2\u2010Catenin Promotes Esophageal Cancer Metastasis by Cooperating With CDK5"],"prefix":"10.1002","volume":"12","author":[{"given":"Shu\u2010Jun","family":"Li","sequence":"first","affiliation":[{"name":"State Key Laboratory of Respiratory Disease Guangdong Provincial Key Laboratory of Protein Modification and Degradation School of Basic Medical Sciences Guangzhou Medical University Guangzhou 511495 China"},{"name":"State Key Laboratory of Respiratory Disease Key Laboratory of Biological Targeting Diagnosis Therapy and Rehabilitation of 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