Kinin and Purine Signaling Contributes to Neuroblastoma Metastasis

Abstract

Bone marrow metastasis occurs in approximately 350,000 patients that annually die in the U.S. alone. In view of the importance of tumor cell migration into the bone marrow, we have here investigated effects of various concentrations of stromal cell-derived factor-1 (SDF-1), bradykinin- and ATP on bone marrow metastasis. We show for first time that bradykinin augmented chemotactic responsiveness of neuroblastoma cells to SDF-1 and ATP concentrations, encountered under physiological conditions. Bradykinin upregulated VEGF expression, increased metalloproteinase activity and induced adhesion of neuroblastoma cells. Bradykinin augmented SDF-1-induced intracellular Ca2+ mobilization as well as resensitization and expression of ATP-sensing P2X7 receptors. Bradykinin treatment resulted in higher gene expression levels of the truncated P2X7B receptor compared to those of the P2X7A full-length isoform. Bradykinin as pro-metastatic factor induced tumor proliferation that was significantly decreased by P2X7 receptor antagonists; however, the peptide did not enhance cell death nor P2X7A receptor-related pore activity, promoting neuroblastoma growth. Furthermore, immunodeficient nude/nude mice transplanted with bradykinin-pretreated neuroblastoma cells revealed significantly higher metastasis rates compared to animals injected with untreated cells. In contrast, animals receiving Brilliant Blue G, a P2X7 receptor antagonist, did not show any specific dissemination of neuroblastoma cells to the bone marrow and liver, and metastasis rates were drastically reduced. Our data suggests correlated actions of kinins and purines in neuroblastoma dissemination, providing novel avenues for clinic research in preventing metastasis.

P2X7 in Cancer: From Molecular Mechanisms to Therapeutics

“…Adinolfi and colleagues first demonstrated the growth-promoting activity of P2X7 in four different tumor models in mice including HEK-293 cells and ACN neuroblastoma derived xenografts and syngeneic colon carcinoma and melanoma models (Adinolfi et al, 2012a). Several studies have confirmed these data and extended them, demonstrating that P2X7 blockade leads to a general reduction of tumor burden in neuroblastoma (Amoroso et al, 2015;Ulrich et al, 2018), mesothelioma (Amoroso et al, 2016), melanoma Adinolfi et al, 2015a), glioma (Ji et al, 2018), osteosarcoma (Zhang et al, 2019a) and myeloid leukemia (De Marchi et al, 2019a).…”

The P2X7 Receptor: Central Hub of Brain Diseases

“…Consistently, overexpression of the P2X7 receptor in a naturally low-expressing human glioma cell line conferred modest in vitro growth advantages, but largely accelerated tumor growth in vivo (Bergamin et al, 2019), reinforcing a trophic role for this receptor. Also, in a mouse model of neuroblastoma, a rare intracranial tumor that affects immature or developing cells of the nervous system, chronic blockade of the P2X7 receptor in tumor-bearing mice diminished progression and metastasis (Ulrich et al, 2018).…”

P2X7 Receptor Upregulation in Huntington’s Disease Brains

“…A pathophysiological role of P2X7R-B has been described in multiple conditions, including bone cancer ( Giuliani et al, 2014 ) and bone differentiation ( Carluccio et al, 2019 ). It has also been described in neural progenitors ( Glaser et al, 2014 ), neuroblastoma cells ( Ulrich et al, 2018 ), and glioblastoma cells ( Ziberi et al, 2019 ). P2X7R-H contains 505 aa and is also known as P2X7R-ΔTM1, since the TM1 is absent.…”