Aggrephagy Reshapes Tumor Microenvironment in Laryngeal Squamous Cell Carcinoma: Insights From Single-Cell Transcriptomics

Abstract

Background

The tumor microenvironment (TME) plays a crucial role in the development and progression of laryngeal squamous cell carcinoma (LSCC). However, the mechanisms by which aggrephagy reshapes the TME remain unclear.

Materials and Methods

Single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing (bulk RNA-seq) data of LSCC were obtained from the TCGA and GEO databases. Seurat and Monocle2 packages were employed for cell clustering and pseudotime trajectory analysis. SCENIC package was used to predict key transcription factors. The CellChat package was used to evaluate intercellular communication, and the GSVA method was applied to calculate signature scores across bulk RNA-seq datasets. The clinical significance of aggrephagy-associated subpopulations was assessed through survival analysis. Immunofluorescence staining was performed to evaluate TUBA1B expression and the infiltration of B cells and T cells in tumor tissues.

Results

The LSCC TME was mostly made up of epithelial cells, T/B lymphocytes, and myeloid cells. The TUBA1B + MaligEpi subpopulation of malignant epithelial cells had the highest level of stemness and was linked to a poor prognosis. The TUBA1B + subset of B cells showed impaired function, while the VIM + subset was able to present antigens. The HSP90AA1+ subset of macrophages demonstrated high inflammatory activity and was linked to TNF/NF-κB signaling. Cell communication analysis showed that VIM+ and TUBA1C + malignant epithelial cells and CD8+ T cells interacted strongly with each other. This could be a way for the immune system to be suppressed. TUBA1B was also highly expressed in several types of cancer and was linked to poor prognosis. B cell and T cell immune infiltration was accompanied by high TUBA1B expression, as confirmed by immunofluorescence.

Conclusion

Aggrephagy is a key driver in TME remodeling in LSCC, highlighting TUBA1B as a promising prognostic biomarker and potential therapeutic target.