Molecular subtypes associated with lymph node metastasis in colorectal cancer reveal the tumor-promoting role of GDI1 and the preclinical activity of deslanoside

Background Lymph node metastasis (LNM) is central to colorectal cancer staging and treatment decisions and marks a critical transition from local growth to regional dissemination. The molecular heterogeneity associated with lymph node metastasis and the corresponding intervention leads require systematic investigation. Methods This study established the C1 to C4 molecular subtypes from lymph node metastasis-related expression features. Multiple transcriptomic cohorts, clinical composition, pathway features, and TIDE, IPS, and ESTIMATE scores were integrated to characterize subtype differences. Analyses across multiple cohorts were used to screen candidate genes associated with progression. Cellular and subcutaneous xenograft experiments were performed to evaluate GDI1 function. The GDI1 structure was used for virtual screening and molecular dynamics analysis, followed by phenotypic evaluation of Deslanoside in cellular and xenograft models. Results Four molecular subtypes displayed distinct expression profiles, clinical composition, and tumor microenvironment features. C4 showed coordinated enrichment of invasive, mesenchymal, hypoxia-related, and stromal programs together with algorithmic features associated with restricted immune effector activity. Analyses across multiple cohorts identified GDI1 as a candidate gene associated with adverse outcomes and progression. Two independent shRNAs reduced GDI1 expression, cell growth, and migration-related phenotypes in two colorectal cancer cell lines. GDI1 knockdown using shGDI1 #1 also reduced subcutaneous xenograft burden. Deslanoside ranked among the leading candidates in the computational screen. Deslanoside treatment produced antitumor phenotypes in cellular and xenograft models that were directionally concordant with those produced by GDI1 depletion. Conclusion This study delineates the molecular heterogeneity associated with lymph node metastasis in colorectal cancer, supports a tumor-promoting role for GDI1, and provides a basis for further investigation of Deslanoside as a candidate compound.