VEGF-induced intracellular Ca2+ oscillations are down-regulated and do not stimulate angiogenesis in breast cancer-derived endothelial colony forming cells

Francesco Lodola, Umberto Laforenza, Fabio Cattaneo, Federico Alessandro Ruffinatti, Valentina Poletto, Margherita Massa, Richard Tancredi, Estella Zuccolo, Dlzar Alì Khdar, Alberto Riccardi, Marco Biggiogera, Vittorio Rosti, Germano Guerra, Francesco Moccia

Research output: Contribution to journalArticle

Abstract

Endothelial colony forming cells (ECFCs) represent a population of truly endothelial precursors that promote the angiogenic switch in solid tumors, such as breast cancer (BC). The intracellular Ca2+ toolkit, which drives the pro-angiogenic response to VEGF, is remodelled in tumor-associated ECFCs such that they are seemingly insensitive to this growth factor. This feature could underlie the relative failure of anti-VEGF therapies in cancer patients. Herein, we investigated whether and how VEGF uses Ca2+ signalling to control angiogenesis in BC-derived ECFCs (BC-ECFCs). Although VEGFR-2 was normally expressed, VEGF failed to induce proliferation and in vitro tubulogenesis in BC-ECFCs. Likewise, VEGF did not trigger robust Ca2+ oscillations in these cells. Similar to normal cells, VEGF-induced intracellular Ca2+ oscillations were triggered by inositol-1,4,5-trisphosphate-dependent Ca2+ release from the endoplasmic reticulum (ER) and maintained by store-operated Ca2+ entry (SOCE). However, InsP3-dependent Ca2+ release was significantly lower in BC-ECFCs due to the down-regulation of ER Ca2+ levels, while there was no remarkable difference in the amplitude, pharmacological profile and molecular composition of SOCE. Thus, the attenuation of the pro-angiogenic Ca2+ response to VEGF was seemingly due to the reduction in ER Ca2+ concentration, which prevents VEGF from triggering robust intracellular Ca2+ oscillations. However, the pharmacological inhibition of SOCE prevented BC-ECFC proliferation and in vitro tubulogenesis. These findings demonstrate for the first time that BC-ECFCs are insensitive to VEGF, which might explain at cellular and molecular levels the failure of anti-VEGF therapies in BC patients, and hint at SOCE as a novel molecular target for this disease.

Original languageEnglish
Pages (from-to)95223-95246
Number of pages24
JournalOncotarget
Volume8
Issue number56
DOIs
Publication statusPublished - Nov 10 2017

    Fingerprint

Keywords

  • Journal Article

Cite this

Lodola, F., Laforenza, U., Cattaneo, F., Ruffinatti, F. A., Poletto, V., Massa, M., Tancredi, R., Zuccolo, E., Khdar, D. A., Riccardi, A., Biggiogera, M., Rosti, V., Guerra, G., & Moccia, F. (2017). VEGF-induced intracellular Ca2+ oscillations are down-regulated and do not stimulate angiogenesis in breast cancer-derived endothelial colony forming cells. Oncotarget, 8(56), 95223-95246. https://doi.org/10.18632/oncotarget.20255