Análise computacional da expressão de correceptores de SARS-CoV-2 em células estromais do microambiente do melanoma Murino B16F10

Abstract

The COVID-19 pandemic highlighted the need to better understand the mechanisms of viral entry into host cells, particularly given the impact of SARS-CoV-2 infection on cancer patients. Although viral entry is primarily associated with the interaction between angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2), studies have demonstrated that cellular coreceptors, molecules that facilitate or modulate viral adhesion, internalization, and dissemination, also play important roles in this process. In the context of cancer, these molecules are involved in biological processes such as cell adhesion, angiogenesis, extracellular matrix remodeling, and tumor progression, particularly within the tumor microenvironment. Therefore, the present study aimed to computationally analyze the expression of SARS-CoV-2 receptors and coreceptors in stromal cells of the murine B16F10 melanoma microenvironment using publicly available single-cell RNA sequencing (scRNA-seq) data. The genes Eca2, Tmprss2, Itgb1, Itgb3, Dpp4, Axl, Hspa5, Bsg, Nrp1, Vim, Hspg2, and Tlr4 were evaluated and normalized to the Actb gene. Data were obtained from the Murine Melanoma Atlas platform and analyzed using FIJI/ImageJ software through custom-developed macros to quantify relative gene expression in expression maps by applying image processing techniques, including grayscale conversion, intensity thresholding, and quantification of the labeled area. In addition to the overall analysis of the tumor microenvironment, a segmented analysis was performed in which cancer-associated fibroblasts (CAFs) and tumor endothelial cells were isolated and analyzed separately, allowing comparison of expression profiles between these stromal cell populations. The results demonstrated low expression of Eca2 and Tmprss2 within the tumor microenvironment, whereas the coreceptors exhibited heterogeneous and cell type-dependent expression patterns. Genes such as Hspa5, Bsg, Vim, and Itgb1 showed widespread expression, whereas Axl, Dpp4, Itgb3, and Tlr4 displayed more restricted expression profiles. In the segmented analysis, Bsg, Vim, Hspa5, Itgb1, and Axl were more highly expressed in CAFs, whereas Vim, Bsg, Itgb1, Hspa5, and Nrp1 were more highly expressed in tumor endothelial cells. Additionally, comparative analysis of neuropilin-1 revealed a high degree of conservation across different species. These findings indicate that SARS-CoV-2 coreceptors exhibit distinct expression patterns within the tumor microenvironment and are associated with biological processes related to stromal activation, angiogenesis, cell–matrix interactions, and tumor progression. The computational characterization of these molecules expands the understanding of their distribution within the melanoma microenvironment and provides a foundation for future experimental investigations into their role in tumor biology and their potential as therapeutic targets.