IADR Abstract Archives
Spatial Transcriptomics for Identifying Biomarkers of Malignant Transformation in Oral Premalignant Lesions
Objectives: Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers worldwide, with high mortality rates due to late-stage diagnosis. A critical challenge in early detection is the incomplete understanding of molecular alterations driving malignant transformation and the lack of reliable biomarkers.
Methods: In this study, we employed spatial transcriptomics and multiplex immunofluorescence (mIF) to characterize molecular and cellular changes associated with OSCC development in a well-characterized cohort of 148 patients with progressive and non-progressive oral premalignant lesions (OPLs). Spatial transcriptomic analysis was performed using Xenium in situ with the Human Immuno-Oncology Profiling Panel, enabling high-resolution mapping of gene expression and cellular interactions within the tumor microenvironment. mIF was utilized to assess immune cell composition, protein expression dynamics, and spatial localization of key tumor-associated markers. Additionally, we analyzed The Cancer Genome Atlas (TCGA) cohort of over 500 head and neck cancer patients and conducted deconvolution analysis for external validation
Results: A total of 15 unique cell clusters were identified, including macrophages, epithelial cells, plasma cells, mast cells, fibroblasts, T cells, endothelial cells, B cells, dendritic cells, and Treg cells. Notably, cancer cases exhibited lower macrophage 1 and epithelial 1 frequencies but an increased plasma cell frequency. Subcluster analysis revealed that CXCL14 high subclusters were significantly elevated in progressive and cancer cases, while ANXA1 high subclusters were reduced in cancer. CCL21 high epithelial subclusters decreased across disease stages, whereas FEZ1 high subclusters increased. Spatial analysis demonstrated a significantly greater distance between T cells and epithelial cells in progressive OPL and OSCC, suggesting immune exclusion. In an independent cohort, reductions in macrophage 1 and epithelial cell 1 frequencies were validated, reinforcing their potential as biomarkers for malignant transformation.
Conclusions: These findings provide novel insights into immune and epithelial alterations in OSCC progression and highlight potential diagnostic biomarkers for early detection.
Methods: In this study, we employed spatial transcriptomics and multiplex immunofluorescence (mIF) to characterize molecular and cellular changes associated with OSCC development in a well-characterized cohort of 148 patients with progressive and non-progressive oral premalignant lesions (OPLs). Spatial transcriptomic analysis was performed using Xenium in situ with the Human Immuno-Oncology Profiling Panel, enabling high-resolution mapping of gene expression and cellular interactions within the tumor microenvironment. mIF was utilized to assess immune cell composition, protein expression dynamics, and spatial localization of key tumor-associated markers. Additionally, we analyzed The Cancer Genome Atlas (TCGA) cohort of over 500 head and neck cancer patients and conducted deconvolution analysis for external validation
Results: A total of 15 unique cell clusters were identified, including macrophages, epithelial cells, plasma cells, mast cells, fibroblasts, T cells, endothelial cells, B cells, dendritic cells, and Treg cells. Notably, cancer cases exhibited lower macrophage 1 and epithelial 1 frequencies but an increased plasma cell frequency. Subcluster analysis revealed that CXCL14 high subclusters were significantly elevated in progressive and cancer cases, while ANXA1 high subclusters were reduced in cancer. CCL21 high epithelial subclusters decreased across disease stages, whereas FEZ1 high subclusters increased. Spatial analysis demonstrated a significantly greater distance between T cells and epithelial cells in progressive OPL and OSCC, suggesting immune exclusion. In an independent cohort, reductions in macrophage 1 and epithelial cell 1 frequencies were validated, reinforcing their potential as biomarkers for malignant transformation.
Conclusions: These findings provide novel insights into immune and epithelial alterations in OSCC progression and highlight potential diagnostic biomarkers for early detection.