Extracellular Vesicle-Mediated Purinergic Signaling Contributes to Host Microenvironment Plasticity and Metastasis in Triple Negative Breast Cancer

• Published in: International journal of molecular sciences 2021-01, Vol.22 (2), p.597
• Main Authors: Duan, Suzann, Nordmeier, Senny, Byrnes, Aidan E, Buxton, Iain L O
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animals; Breast cancer; Cell Line; Cell Line, Tumor; Cell Movement; Endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - pathology; Endothelium; Enzymes; Evaluation; exosomes; Extracellular vesicles; Extracellular Vesicles - metabolism; Extracellular Vesicles - pathology; Female; Flow cytometry; Humans; Kinases; Labeling; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Lung Neoplasms - secondary; Lungs; Metastases; Metastasis; Mice, SCID; Microenvironments; NM23 Nucleoside Diphosphate Kinases - metabolism; Nucleoside-diphosphate kinase; Phosphotransferase; Proteins; purinergic signaling; Receptors, Purinergic P2Y - metabolism; Signal Transduction; Triple Negative Breast Neoplasms - metabolism; Triple Negative Breast Neoplasms - pathology; Tumor Microenvironment
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22020597

Metastasis accounts for over 90% of cancer-related deaths, yet the mechanisms guiding this process remain unclear. Secreted nucleoside diphosphate kinase A and B (NDPK) support breast cancer metastasis. Proteomic evidence confirms their presence in breast cancer-derived extracellular vesicles (EVs). We investigated the role of EV-associated NDPK in modulating the host microenvironment in favor of pre-metastatic niche formation. We measured NDPK expression and activity in EVs isolated from triple-negative breast cancer (MDA-MB-231) and non-tumorigenic mammary epithelial (HME1) cells using flow cytometry, western blot, and ATP assay. We evaluated the effects of EV-associated NDPK on endothelial cell migration, vascular remodeling, and metastasis. We further assessed MDA-MB-231 EV-induced proteomic changes in support of pre-metastatic lung niche formation. NDPK-B expression and phosphotransferase activity were enriched in MDA-MB-231 EVs that promote vascular endothelial cell migration and disrupt monolayer integrity. MDA-MB-231 EV-treated mice demonstrate pulmonary vascular leakage and enhanced experimental lung metastasis, whereas treatment with an NDPK inhibitor or a P2Y1 purinoreceptor antagonist blunts these effects. We identified perturbations to the purinergic signaling pathway in experimental lungs, lending evidence to support a role for EV-associated NDPK-B in lung pre-metastatic niche formation and metastatic outgrowth. These studies prompt further evaluation of NDPK-mediated EV signaling using targeted genetic silencing approaches.