Single-Cell and Spatial Analysis Reveal the Differences Between Left-Sided and Right-Sided Colorectal Cancer

Colorectal cancer (CRC) exhibits heterogeneity based on tumour laterality, with right-sided tumours demonstrating distinct genetic, immunogenic and clinical behaviours compared to left-sided counterparts. In this study, we conducted single-cell RNA sequencing and spatial transcriptomics to dissect the distinct molecular landscapes of left- and right-sided CRC. We identified enhanced germinal centre B cell infiltration and CXCL13⁺ T cell enrichment, features linked to adaptive immune priming and better immunotherapy responsiveness. Right-sided CRC epithelial cells exhibited proliferative and immunogenic phenotypes, marked by upregulated immune-related pathways and chemokine-driven interactions with lymphocytes. Spatial analysis revealed organized tertiary lymphoid structure-immune microenvironment crosstalk in right-sided tumours, mediated by CCL19/21-CCR7 signalling. In contrast, left-sided CRC tumours displayed stromal-epithelial interactions favouring angiogenesis and metabolic reprogramming. Our findings established a laterality-specific immune microenvironment in CRC, providing insights for precise therapeutic strategies of left- and right-sided CRC.