ATP confers tumorigenic properties to dendritic cells by inducing amphiregulin secretion

Abstract

IntroductionThe interactions between the immune system and angiogenesis are determinant for the regulation of tumor growth. The mechanisms of these interactions are not well understood at this time. Dendritic cells (DCs) are known to play a central role in these interactions by releasing vascular endothelial growth factor-A (VEGF-A). 1,2 VEGF-A is able to induce endothelial-like differentiation of tumor-infiltrating precursors of DCs and their migration to vessels to participate to vasculogenesis. 3 VEGF-A was also reported to inhibit the function of mature DCs. 4 Immune cells such as tumor-associated macrophages and T cells were also reported to regulate angiogenesis through the secretion of potent angiogenic mediators such as angiopoietin-2 and interleukin-17 . 5,6 We demonstrated previously that ATP up-regulates the expression by monocyte-derived DCs (MoDCs) of numerous genes that may play a role in immunosuppression, in particular thrombospondin-1 and indoleamine 2,3-dioxygenase. 2,7 Some epidermal growth factor receptor (EGFR) ligands were also ATP target genes in MoDCs, in particular, amphiregulin (AREG), which was the most highly up-regulated gene. AREG is a cell type-dependent mitogenic factor that binds to ErbB1 receptor, also called EGFR. 8 AREG is implicated in tumorigenesis and angiogenesis. 9,10 Ma et al showed a reduction of the tumor mass and the intratumoral vascularization using an antisense cDNA of AREG in a breast cell line. 11 A clinical study showed that there was a significant overexpression of AREG and VEGF in primary breast cancer. 12 Kato et al showed that AREG can induce the proliferation of rat vascular smooth muscle cells and could be implicated in arterial remodeling. 13 We demonstrated here for the first time that human and mouse DCs are a source of AREG, an EGFR ligand with tumorigenic properties. Methods MiceMale 6-to 8-week-old C57BL/6 mice were obtained from Charles River Laboratories. All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals as adopted and promulgated by the US National Institutes of Health (NIH) and were approved by the Comission d'Ethique et du Bien-Etre Animal (CEBEA) ethical committee. ReagentsATP, adenosine 5Ј-O-(3-thiotriphosphate; ATP␥S), UTP, prostaglandin E2 (PGE 2 ), adenosine (Ado), NECA (5Ј-(N-ethylcarboxamido)adenosine), suramin, MRS1754, and lipopolysaccharide (LPS) were obtained from Sigma-Aldrich. Monoclonal anti-human and anti-mouse AREG blocking antibodies were purchased at R&D Systems. Preparation of human monocyte-derived DCsPeripheral blood mononuclear cells were isolated from leukocyte-enriched buffy coats of healthy volunteer donors by standard density gradient centrifugation using Lymphoprep solution from Nycomed. Mononuclear cells (2.5 ϫ 10 8 ) were allowed to adhere during 1 hour and 30 minutes at 37°C at 5% CO 2 in 75-cm 2 cell culture flasks. Nonadherent cells were removed, and adherent cells were cultured in 15 mL of culture medium For personal use only. on April 4, 2019. by guest ...

Lung Tumor-Associated Dendritic Cell-Derived Amphiregulin Increased Cancer Progression

“…Amphiregulin also elevates angiogenic activity, promoting tumor growth and metastasis in pancreatic, colorectal, liver, and lung cancers (9)(10)(11)(12). A recent study has indicated that cancer-derived ATP increased the expression of amphiregulin in DCs in a mouse lung cancer cell line (13). Patients with high levels of amphiregulin have poorer prognoses and higher resistance than patients with low levels of amphiregulin (14).…”

Roles of extracellular nucleotides and P2 receptors in ectodomain shedding

“…UTP also induced amphiregulin shedding from LPS-stimulated murine bone marrow-derived, but not from human monocyte-derived, dendritic cells [78]. Moreover, suramin inhibited ATPγS-induced amphiregulin shedding from human monocyte-derived dendritic cells [78], but the specific P2 receptors involved were not identified.…”

“…UTP also induced amphiregulin shedding from LPS-stimulated murine bone marrow-derived, but not from human monocyte-derived, dendritic cells [78]. Moreover, suramin inhibited ATPγS-induced amphiregulin shedding from human monocyte-derived dendritic cells [78], but the specific P2 receptors involved were not identified. Notably, amphiregulin released from murine bone marrow-derived dendritic cells stimulated tumour growth in vivo [78] indicating that nucleotides can confer tumourigenic properties to dendritic cells by inducing amphiregulin release.…”

“…Moreover, suramin inhibited ATPγS-induced amphiregulin shedding from human monocyte-derived dendritic cells [78], but the specific P2 receptors involved were not identified. Notably, amphiregulin released from murine bone marrow-derived dendritic cells stimulated tumour growth in vivo [78] indicating that nucleotides can confer tumourigenic properties to dendritic cells by inducing amphiregulin release. This contrasts the anti-tumourigenic properties of dendritic cells following P2X7 activation and subsequent NLRP3 inflammasome stimulation to promote anti-tumour immunity [79].…”

“…ATP and ATPγS induced slow (24 hours) shedding of amphiregulin from LPS-stimulated human monocyte-derived and murine bone marrow-derived dendritic cells [78]. UTP also induced amphiregulin shedding from LPS-stimulated murine bone marrow-derived, but not from human monocyte-derived, dendritic cells [78].…”