Induction of heparanase via IL-10 correlates with a high infiltration of CD163+ M2-type tumor-associated macrophages in inflammatory breast carcinomas

Abstract

Inflammatory breast cancer (IBC) is the most aggressive and lethal form of breast cancer, characterized by a high infiltration of tumor-associated macrophages and poor prognosis. To identify new biomarkers and to elucidate the molecular mechanisms underlying IBC pathogenesis, we investigated the expression pattern of heparanase (HPSE) and its activator cathepsin L (CTSL). First, we quantitated the HPSE and CTSL mRNA levels in a cohort of breast cancer patients after curative surgery (20 IBC and 20-non-IBC). We discovered that both HPSE and CTSL mRNA levels were significantly induced in IBC tissue vis-à-vis non-IBC patients ( p <0 .05 and p <0 .001, respectively). According to the molecular subtypes, HPSE mRNA levels were significantly higher in carcinoma tissues of triple negative (TN)-IBC as compared to TN-non-IBC ( p <0 .05). Mechanistically, we discovered that pharmacological inhibition of HPSE activity resulted in a significant reduction of invasiveness in the IBC SUM149 cell line. Moreover, siRNA-mediated HPSE knockdown significantly downregulated the expression of the metastasis-related gene MMP2 and the cancer stem cell marker CD44. We also found that IBC tumors revealed robust heparanase immune-reactivity and CD163+ M2-type tumor-associated macrophages, with a positive correlation of both markers. Moreover, the secretome of axillary tributaries blood IBC CD14+ monocytes and the cytokine IL-10 significantly upregulated HPSE mRNA and protein expression in SUM149 cells. Intriguingly, massively elevated IL-10 mRNA expression with a trend of positive correlation with HPSE mRNA expression was detected in carcinoma tissue of IBC. Our findings highlight a possible role played by CD14+ monocytes and CD163+ M2-type tumor-associated macrophages in regulating HPSE expression possibly via IL-10. Overall, we suggest that heparanase, cathepsin L and CD14+ monocytes-derived IL-10 may play an important role in the pathogenesis of IBC and their targeting could have therapeutic implications.

Inflammatory Breast Carcinoma: Elevated microRNA miR-181b-5p and Reduced miR-200b-3p, miR-200c-3p, and miR-203a-3p Expression as Potential Biomarkers with Diagnostic Value

“…Interestingly, hub genes in module 4 were implicated in different types of cancer as well as signaling pathways associated with cancer, including proteoglycans, and human papillomavirus infection. This conforms with our previous study reporting that triple-negative IBC tumors overexpress the heparan sulfate proteoglycan Syndecan-1 [ 43 ], the substrate whose functional activity is affected by the heparanase enzyme, which is also overexpressed in tumors of triple-negative IBC [ 44 ]. Further, we and others have previously shown that IBC tumors confer multiple viral DNA prevalence [ 45 ] and that viral DNA is significantly associated with the triple-negative IBC, the most aggressive phenotype [ 46 ].…”

“…One possible caveat that could be associated with this study is the quantification of the expression levels of dysregulated miRNAs in total carcinoma tissue lysates of IBC and non-IBC, which may overlap with miRNAs derived from the infiltrated immune cells, particularly macrophages, as we and others have shown that IBC tumors are known to be a highly infiltrated with CD163+ M2-type tumor-associated macrophages [ 44 ]. For example, while miR-200b expression was downregulated in our triple-negative breast cancer cell lines compared to the ER+ luminal cell line MCF-7, it was expressed at a higher level in the IBC cell line SUM-149 compared to the non-IBC triple-negative breast cancer cell line MDA-MB-231.…”

Knockdown of the prognostic cancer stem cell marker Musashi-1 decreases radio-resistance while enhancing apoptosis in hormone receptor-positive breast cancer cells via p21WAF1/CIP1

“…MTT assays were performed as previously described (El-Nadi et al 2020 ). Briefly, 4000 cells/well of MSI-1 knockdown and control cells were seeded in 96-well plates 48 h after transfection and cultured for an additional 24 h. Then, chemotherapy was added: specifically, varying doses of cisplatin, and doxorubicin were used.…”

Syndecan-1 Promotes Angiogenesis in Triple-Negative Breast Cancer through the Prognostically Relevant Tissue Factor Pathway and Additional Angiogenic Routes

“…The non-enzymatic activity of heparanase—known to be modulated by Sdc-1 expression—induces F3 expression in cancer cells and impacts cancer progression [ 44 , 45 ]. We have recently discovered an overexpression of heparanase and Sdc-1 in triple-negative inflammatory breast cancer, an aggressive form of breast cancer represented by the SUM-149 cell line employed in this study [ 14 , 46 ]. F3 promotes angiogenesis of the microvascular endothelial cells [ 47 ] and the overexpression of tumor F3-induced increased angiogenesis enables human melanoma xenografts to form lung metastasis [ 48 ].…”