M2 macrophages induce ovarian cancer cell proliferation via a heparin binding epidermal growth factor/matrix metalloproteinase 9 intercellular feedback loop

Abstract

In ovarian cancer, a high ratio of anti-inflammatory M2 to pro-inflammatory M1 macrophages correlates with poor patient prognosis. The mechanisms driving poor tumor outcome as a result of the presence of M2 macrophages in the tumor microenvironment remain unclear and are challenging to study with current techniques. Therefore, in this study we utilized a micro-culture device previously developed by our lab to model concentrated paracrine signaling in order to address our hypothesis that interactions between M2 macrophages and ovarian cancer cells induce tumor cell proliferation. Using the micro-culture device, we determined that co-culture with M2-differentiated primary macrophages or THP-1 increased OVCA433 proliferation by 10–12%. This effect was eliminated with epidermal growth factor receptor (EGFR) or heparin-bound epidermal growth factor (HB-EGF) neutralizing antibodies and HBEGF expression in peripheral blood mononuclear cells from ovarian cancer patients was 9-fold higher than healthy individuals, suggesting a role for HB-EGF in tumor progression. However, addition of HB-EGF at levels secreted by macrophages or macrophage-conditioned media did not induce proliferation to the same extent, indicating a role for other factors in this process. Matrix metalloproteinase-9, MMP-9, which cleaves membrane-bound HB-EGF, was elevated in co-culture and its inhibition decreased proliferation. Utilizing inhibitors and siRNA against MMP9 in each population, we determined that macrophage-secreted MMP-9 released HB-EGF from macrophages, which increased MMP9 in OVCA433, resulting in a positive feedback loop to drive HB-EGF release and increase proliferation in co-culture. Identification of multi-cellular interactions such as this may provide insight into how to most effectively control ovarian cancer progression.

Upregulation of IGF1 by tumor-associated macrophages promotes the proliferation and migration of epithelial ovarian cancer cells

“…The TME is a supportive and receptive tissue microenvironment undergoing a series of molecular and cellular changes to form metastatic-designated sites, or the fertile soil in preparation for metastatic tumor cell seed colonization, thus supporting tumor settlement in distant organs and promoting tumor metastasis (6)(7)(8). As the most common immune cells in the TME, TAMs infiltrate the TME in OC in large numbers and play an important role in the formation of the OC TME and education of OC cells to develop to become more malignant by secreting various factors, such as IL-6 and IDO (16,18,19,39,40). Indeed, therapeutic methods targeting TAMs have been demonstrated as effective for controlling OC.…”

“…In addition to the microenvironment within tumor tissues, TAMs are also distributed in some special organs and lymph nodes, associated with metastasis to these regions (13,14). TAMs infiltrate into OC tissues in large numbers (15)(16)(17), contributing to the progression of OC (18,19), thus negatively affecting the progression-free survival rates and overall survival rates of these patients (16). However, the precise mechanism of how TAMs contribute to the progression of OC remains unclear.…”

KRAS mutation and epithelial–macrophage interplay in pancreatic neoplastic transformation

“…Tumorassociated macrophages can secrete epidermal growth factor (EGF), which activates epithelial EGFR pathway, and exerts protumorigenic effects on the neoplastic epithelium. 31,32 We first screened EGF and EGFR expression in our co-cultured macrophages and pancreas organoids (WT) by quantitative PCR. EGF was highly expressed in macrophages, but not in epithelial organoids, whereas EGFR was highly expressed in epithelial organoids ( Fig.…”

s-HBEGF/SIRT1 circuit-dictated crosstalk between vascular endothelial cells and keratinocytes mediates sorafenib-induced hand–foot skin reaction that can be reversed by nicotinamide

“…It has been reported that enzymes matrix metalloproteinases 9 (MMP9) and matrix metalloproteinases 3 (MMP3) are involved in the cleavage of pro-HBEGF to form s-HBEGF. 37,38 Interestingly, we also observed that sorafenib could increase both MMP9 and MMP3 protein levels in a transcription-dependent manner (Supplementary information, Fig. S2a, b).…”