biostudies-literatureUnknown9(6)He Z<h4>Background</h4>Gallbladder cancer (GBC) is a rare but aggressive malignancy, often detected late due to early asymptomatic stages. Understanding cellular and molecular changes from normal tissue to high-grade intraepithelial neoplasia (HGIN) and invasive GBC is vital for identifying early biomarkers and therapeutic targets.<h4>Methods</h4>We performed single-cell RNA sequencing on 98,113 cells derived from 2 normal adjacent tissues (NAT), 2 HGIN, and 6 GBC samples. The cellular diversity and heterogeneity, particularly within epithelial and immune cell populations in NAT-HGIN-GBC, were investigated utilizing single-cell RNA sequencing, bulk RNA sequencing (bulk RNA-seq), and 10 machine learning methodologies. Furthermore, the intercellular crosstalk between epithelial cells and tumor iHepatology communicationse0697https://www.ebi.ac.uk/biostudies/studies/S-EPMC12088638biostudies-literatureSingle-cell analyses unravel ecosystem dynamics and intercellular crosstalk during gallbladder cancer malignant transformation.PMC12088638He ZCao JPeng CWang XGao HYang SYu YDi ZfalseSingle-cell analyses unravel ecosystem dynamics and intercellular crosstalk during gallbladder cancer malignant transformation.<h4>Background</h4>Gallbladder cancer (GBC) is a rare but aggressive malignancy, often detected late due to early asymptomatic stages. Understanding cellular and molecular changes from normal tissue to high-grade intraepithelial neoplasia (HGIN) and invasive GBC is vital for identifying early biomarkers and therapeutic targets.<h4>Methods</h4>We performed single-cell RNA sequencing on 98,113 cells derived from 2 normal adjacent tissues (NAT), 2 HGIN, and 6 GBC samples. The cellular diversity and heterogeneity, particularly within epithelial and immune cell populations in NAT-HGIN-GBC, were investigated utilizing single-cell RNA sequencing, bulk RNA sequencing (bulk RNA-seq), and 10 machine learning methodologies. Furthermore, the intercellular crosstalk between epithelial cells and tumor i2025-01-01T00:00:00Z2025 Jun2026-06-07T05:29:22.328Z2026-06-07T03:07:23.597ZS-EPMC120886384037748410.1097/HC9.0000000000000697