Dissection of paracrine/autocrine interplay in lung tumor microenvironment mimicking cancer cell-monocyte co-culture models reveals proteins that promote inflammation and metastasis

Abstract

Background Tumor cell-monocyte interactions play crucial roles in shaping up the pro-tumorigenic phenotype and functional output of tumor-associated macrophages. Within the tumor microenvironment, such heterotypic cell–cell interactions are known to occur via secretory proteins. Secretory proteins establish a diabolic liaison between tumor cells and monocytes, leading to their recruitment, subsequent polarization and consequent tumor progression. Methods We co-cultured model lung adenocarcinoma cell line A549 with model monocytes, THP-1 to delineate the interactions between them. The levels of prototypical pro-inflammatory cytokines like TNF-𝛼, IL-6 and anti-inflammatory cytokines like IL-10 were measured by ELISA. Migration, invasion and attachment independence of lung cancer cells was assessed by wound healing, transwell invasion and colony formation assays respectively. The status of EMT was evaluated by immunofluorescence. Identification of secretory proteins differentially expressed in monocultures and co-culture was carried out using SILAC LC–MS/MS. Various insilico tools like Cytoscape, Reacfoam, CHAT and Kaplan–Meier plotter were utilized for association studies, pathway analysis, functional classification, cancer hallmark relevance and predicting the prognostic potential of the candidate secretory proteins respectively. Results Co-culture of A549 and THP-1 cells in 1:10 ratio showed early release of prototypical pro-inflammatory cytokines TNF-𝛼 and IL-6, however anti-inflammatory cytokine, IL-10 was observed to be released at the highest time point. The conditioned medium obtained from this co-culture ratio promoted the migration, invasion and colony formation as well as the EMT of A549 cells. Co-culturing of A549 with THP-1 cells modulated the secretion of proteins involved in cell proliferation, migration, invasion, EMT, inflammation, angiogenesis and inhibition of apoptosis. Among these proteins Versican, Tetranectin, IGFBP2, TUBB4B, C2 and IFI30 were found to correlate with the inflammatory and pro-metastatic milieu observed in our experimental setup. Furthermore, dysregulated expression of these proteins was found to be associated with poor prognosis and negative disease outcomes in lung adenocarcinoma compared to other cancer types. Pharmacological interventions targeting these proteins may serve as useful therapeutic approaches in lung adenocarcinoma. Conclusion In this study, we have demonstrated that the lung cancer cell-monocyte cross-talk modulates the secretion of IFI30, RNH1, CLEC3B, VCAN, IGFBP2, C2 and TUBB4B favoring tumor growth and metastasis.

CD44v5 domain regulates crosstalk between TNBC cells and tumor‐associated macrophages by enhancing the IL‐4R/STAT3 axis

“…IL‐6 level is associated with poor survival rates in various cancers, 32 and IL‐6 promotes breast cancer metastasis, angiogenesis, and tumor escape from immune surveillance. 33 , 34 In our experiments, we also found a high expression of IL‐6 in TNBC, and the secretion of IL‐6 was significantly reduced when CD44v5 antibodies were used.…”

Targets in the Tumour Matrisome to Promote Cancer Therapy Response

“…Versican appears in several poor outcome matrix signatures, including the Matrix Index and C-ECM [8,25]. Versican interacts with many of the ECM components addressed in this review, including HA, FN1, and MMPs, and several studies have delineated roles in the modulation of myeloid cells and inflammation [207,224]. The versikine studies de-scribed above indicate a positive functional impact on the ability of DCs to prime adaptive immunity and promote ICB response.…”

Persistent lung inflammation and alveolar-bronchiolization due to Notch signaling dysregulation in SARS-CoV-2 infected hamster

“…Tubb4b overexpression is reported in several cancers (36). Recent studies implicated that Tubb4b is essential for cancer stem cell niche maintenance via cooperation with Ephrin-B1 and Tubb4b promotes tumor growth and metastasis through lung cancer cells and monocyte crosstalk (36,54). Studies also demonstrate upregulation of Mlf1 in lung SCC (31), elevation of Perp (32) and Grb14 in lung adenocarcinoma (34), and increased Muc1 expression in lung cancer and precancerous lesions (33).…”