Keratin 17 Defines an Epithelial-Mesenchymal Transition-Associated Migratory Cell State in Pancreatic Ductal Adenocarcinoma

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is characterized by early invasion, metastasis, and therapeutic resistance that contribute substantially to its poor clinical outcome. Keratin 17 (K17) is a recognized marker of the aggressive basal-like subtype of PDAC and an independent predictor of adverse prognosis; however, the biological basis for this association remains incompletely understood. To define K17-associated cellular programs, we integrated single-cell RNA sequencing (scRNA-seq) data from 42 patients, Clinical Proteomic Tumor Analysis Consortium (CPTAC) proteomic data from 140 tumors, MACSima multiplex spatial imaging of 63 patients, dual-plex immunohistochemistry of 60 cases, and three-dimensional spheroid migration assays. Single-cell transcriptomic analysis demonstrated significant enrichment of epithelial-mesenchymal transition (EMT)-associated gene programs among K17-positive malignant epithelial cells. Consistent with this finding, K17-positive tumors in the CPTAC cohort exhibited enrichment of the Hallmark EMT pathway at the protein level. Among six candidate markers identified through transcriptomic analyses, laminin subunit gamma-2 (LAMC2) was the only candidate associated with K17 at every validation step. Tumors positive for both K17 and LAMC2 had significantly worse overall survival than double-negative tumors (adjusted hazard ratio = 2.53). Multiplex spatial imaging localized K17-positive cells predominantly to poorly differentiated tumor regions and demonstrated frequent single-cell co-expression with LAMC2. Whole-slide dual-plex immunohistochemistry further confirmed the association between K17 and LAMC2 and showed progressive enrichment of K17/LAMC2-positive tumor cells with increasing intratumoral histologic grade. Functionally, gain- and loss-of-function studies in three-dimensional PDAC spheroids demonstrated that K17 promotes migratory outgrowth, and differences in proliferation did not account for this effect. Collectively, these findings identify K17 as a marker of a poorly differentiated, EMT-associated migratory cell state in PDAC and provide functional evidence that K17 contributes to tumor-cell migration.