In Situ Overexpression of Matricellular Mechanical Proteins Demands Functional Immune Signature and Mitigates Non-Small Cell Lung Cancer Progression

Abstract

Non-small cell lung carcinoma (NSCLC) is a complex cancer biome composed of malignant cells embedded in a sophisticated tumor microenvironment (TME) combined with different initiating cell types, including immune cells and cancer-associated fibroblasts (CAFs), and extracellular matrix (ECM) proteins. However, little is known about these tumors’ immune-matricellular relationship as functional and mechanical barriers. This study investigated 120 patients with NSCLC to describe the immune-matricellular phenotypes of their TME and their relationship with malignant cells. Immunohistochemistry (IHC) was performed to characterize immune checkpoints (PD-L1, LAG-3, CTLA-4+, VISTA 1), T cells (CD3+), cytotoxic T cells (CD8+, Granzyme B), macrophages (CD68+), regulatory T cells (FOXP3+, CD4+), natural killer cells (CD57+), and B lymphocytes (CD20+), whereas CAFs and collagen types I, III, and V were characterized by immunofluorescence (IF). We observed two distinct functional immune-cellular barriers—the first of which showed proximity between malignant cells and cytotoxic T cells, and the second of which showed distant proximity between non-cohesive nests of malignant cells and regulatory T cells. We also identified three tumor-associated matricellular barriers: the first, with a localized increase in CAFs and a low deposition of Col V, the second with increased CAFs, Col III and Col I fibers, and the third with a high amount of Col fibers and CAFs bundled and aligned perpendicularly to the tumor border. The Cox regression analysis was designed in two steps. First, we investigated the relationship between the immune-matricellular components and tumor pathological stage (I, II, and IIIA), and better survival rates were seen in patients whose tumors expressed collagen type III > 24.89 fibers/mm². Then, we included patients who had progressed to pathological stage IV and found an association between poor survival and tumor VISTA 1 expression > 52.86 cells/mm² and CD3+ ≤ 278.5 cells/mm². We thus concluded that differential patterns in the distribution of immune-matricellular phenotypes in the TME of NSCLC patients could be used in translational studies to predict new treatment strategies and improve patient outcome. These data raise the possibility that proteins with mechanical barrier function in NSCLC may be used by cancer cells to protect them from immune cell infiltration and immune-mediated destruction, which can otherwise be targeted effectively with immunotherapy or collagen therapy.

The entanglement of extracellular matrix molecules and immune checkpoint inhibitors in cancer: a systematic review of the literature

“…Similar results were obtained by Yaegashi et al. in non-small cell lung carcinoma (NSCLC) ( 39 ). Yaegashi and colleagues identified three types of ECM barriers in NSCLC: the first represented by a low deposition of collagen V, the second showing an increase of collagen III and collagen I, and the third characterized by a high amount of collagen I fibers perpendicularly aligned to the tumor border.…”

Prognostic value of tumor-associated regulatory T-cells as a biomarker in non-small cell lung cancer: a systematic review and meta-analysis

“…Eventually, 19 studies (16 from manual search and three from crossreferences) were included in the review (Fig. 1) [34,35,[44][45][46][47][48][49][50][51][52][36][37][38][39][40][41][42][43].…”

Clinical Application Perspectives of Lung Cancers 3D Tumor Microenvironment Models for In Vitro Cultures

“…There were also two types of immune cellular barrier identified, which correlated with the type of matricellular tumor barrier. Collagen I/III-CAFs barrier was correlated with a higher presence of CD3 + and CD8 + T cells within high expression of PD-L1 and CTLA-4, whereas collagen V-CAFs barrier was associated with regulatory T cell presence and exhausted CD8 + T cell within production of immunosuppressives factors [65]. The immune synapse formation between antigen presenting cells (APCs) and lymphocyte is a key step for anti-tumor immune response activation.…”