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

Abstract

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.

A six-gene signature related with tumor mutation burden for predicting lymph node metastasis in breast cancer

“…The membrane of lysosome and vacuole, vesicle localization and GTPase regulator activity were mainly enriched in GO analysis, while calcium signaling pathway and salmonella infection were mainly enriched in KEGG analysis. The roles of vesicle localization, calcium signaling and salmonella infection in regulating BC metastasis have been explored in some researches ( 35 - 37 ). Furthermore, we established the TMB-related signature of six genes, including BAHD1, PPM1A, PQLC3, SMPD3, EEF1A1 and S100B, via univariate and multivariate Logistic regression analysis.…”

ATP and cancer immunosurveillance

“…Similar findings have been obtained with human triple-negative breast cancer (TNBC) MDA-MB-231 cells upon the ATP-dependent activation of the transcription factor SRY-box transcription factor 9 (SOX9) (Yang et al, 2020a). Finally, NME/NM23 nucleoside diphosphate kinase 1 (NME1, best known as NDPK-A) and NME2 (best known as NDPK-B) expression on extracellular vesicles from MDA-MB-231 cells reportedly support the formation of pulmonary metastatic niches as a consequence of extracellular ATP generation in situ and consequent activation of purinergic receptor P2Y1 (P2RY1) (Duan et al, 2021).…”

Pre-metastatic Niche Formation in Different Organs Induced by Tumor Extracellular Vesicles

“…Vascular endothelial growth factor-A (VEGF-A) is carried by EVs derived from glioblastoma stem-like cells, increasing vascular permeability in vivo and the angiogenic potential of human brain endothelial cells ( Treps et al, 2017 ). In addition, nucleoside diphosphate kinase B enriched in EVs derived from triple negative breast cancer cells (MDA-MB-231) enhances pulmonary blood vessel leakage and experimental lung metastasis ( Duan et al, 2021 ).…”