Myofibroblasts Derived Type V Collagen Promoting Tissue Mechanical Stress and Facilitating Metastasis and Therapy Resistance of Lung Adenocarcinoma Cells

Abstract Background Lung cancer accounts for 11. 6% of new diagnosed malignancy cases and stands as a major cause of global cancer-related deaths, with a 5-year survival rate of 5% for Lung Adenocarcinoma (LUAD), which is one of the major histology type of lung cancer. Mechanical alterations in the tumor microenvironment, including extracellular matrix (ECM) remodeling and fibroblast activity, leading to changes in tissue mechanical characteristics and play a crucial role in cancer advancement and metastasis. The basement membrane (BM), as a distinct type of ECM, is crucial to maintain the mechanical stress of the tissues but how the BM impact the ECM mechanical characteristics of LUAD is not quite understood. This study delves into identifying the BM genes that impact the internal mechanical stress of tumors, elucidating the effect that mechanical properties influence LUAD metastasis and therapy resistance, and the strategies to reverse these effects. Methods Matrigel was overlaid to the surface of LUAD cells and the migration and invasive ability of LUAD cells were evualated by Transwell and Scratch wound-healing assays. A comprehensive approach based on machine learning was employed to construct the SVMScore, a model built using relevant basement membrane (BM) genes. The analysis of single-cell sequencing data was used to reveal the relationship between SVMScore and the secretion of type V collagen by myofibroblasts. Furthermore, atomic force microscopy was utilized to measure the surface hardness of LUAD tissues, exploring the correlation between SVMScore and type V collagen in relation to the hardness of LUAD tissues. Finally, tumor organoids derived from LUAD patients were co-cultured with myofibroblasts to investigate the effect of type V collagen from myofibroblast to tumor metastasis and therapy resistance of LUAD cells. Results Mechanical stress, simulated by matrix application, enhanced LUAD cell migration and invasion ability, correlating with ECM alterations and EMT pathway activation. SVMScore predicted LUAD patient prognosis and EMT propensity across multiple datasets, revealing its robust prognostic capabilities. Lower SVMScores were associated with worse survival outcomes, increased cancer-related pathways, higher Tumor Mutation Burden and higher internal mechanical stress of LUAD tissues. SVMScore was also predominant with myofibroblast-related COL5A1, a key marker for mechanical stress. The expression of COL5A1 of myofibroblasts influences the SVMScore. COL5A1 from myofibroblasts increases tumor invasiveness and upregulates the EMT pathway of tumor cells. Patient tissues with low SVMScores exhibited higher COL5A1 expression, enhanced EMT propensity, and increased internal mechanical stress. Sorafenib, which can decreases the expression of collagen and fibronectin genes, was used and attenuates the tumor-promoting effect of COL5A1 from myofibroblast, inhibiting proliferation and migration of LUAD cells, and rendering LUAD cells more sensitive to chemotherapy. Conclusions This comprehensive study unveils the intricate relationship between mechanical stress, ECM alterations, and LUAD progression. SVMScore emerges as a potent prognostic tool, reflects tumor mechanical characteristics. Sorafenib intervention targeting COL5A1 secretion provides a potential therapeutic strategy to mitigate LUAD aggressiveness. These findings contribute to a deeper understanding of the biomechanical aspects of LUAD, offering insights for future research and clinical applications. 新摘要 Lung cancer is a leading cause of cancer-related mortality globally, with a dismal 5-year survival rate, particularly for Lung Adenocarcinoma (LUAD). Mechanical changes within the tumor microenvironment, such as extracellular matrix (ECM) remodeling and fibroblast activity, play pivotal roles in cancer progression and metastasis. However, the specific impact of the basement membrane (BM) on the mechanical characteristics of LUAD remains unclear. This study aims to identify BM genes influencing internal mechanical stress in tumors, elucidating their effects on LUAD metastasis and therapy resistance, and exploring strategies to counteract these effects.