Exosomal EPHA2 derived from highly metastatic breast cancer cells promotes angiogenesis by activating the AMPK signaling pathway through Ephrin A1-EPHA2 forward signaling

Abstract

Rationale: Angiogenesis is a fundamental process of tumorigenesis, growth, invasion and metastatic spread. Extracellular vesicles, especially exosomes, released by primary tumors promote angiogenesis and cancer progression. However, the mechanism underlying the pro-angiogenic potency of cancer cell-derived exosomes remains poorly understood. Methods: Exosomes were isolated from breast cancer cells with high metastatic potential (HM) and low metastatic potential (LM). The pro-angiogenic effects of these exosomes were evaluated by in vitro tube formation assays, wound healing assays, rat arterial ring budding assays and in vivo Matrigel plug assays. Subsequently, RNA sequencing, shRNA-mediated gene knockdown, overexpression of different EPHA2 mutants, and small-molecule inhibitors were used to analyze the angiogenesis-promoting effect of exosomal EPHA2 and its potential downstream mechanism. Finally, xenograft tumor models were established using tumor cells expressing different levels of EPHA2 to mimic the secretion of exosomes by tumor cells in vivo, and the metastasis of cancer cells were monitored using the IVIS Spectrum imaging system and Computed Tomography. Results: Herein, we demonstrated that exosomes produced by HM breast cancer cells can promote angiogenesis and metastasis. EPHA2 was rich in HM-derived exosomes and conferred the pro-angiogenic effect. Exosomal EPHA2 can be transferred from HM breast cancer cells to endothelial cells. Moreover, it can stimulate the migration and tube-forming abilities of endothelial cells in vitro and promote angiogenesis and tumor metastasis in vivo. Mechanistically, exosomal EPHA2 activates the AMPK signaling via the ligand Ephrin A1-dependent canonical forward signaling pathway. Moreover, inhibition of the AMPK signaling impairs exosomal EPHA2-mediated pro-angiogenic effects. Conclusion: Our findings identify a novel mechanism of exosomal EPHA2-mediated intercellular communication from breast cancer cells to endothelial cells in the tumor microenvironment to provoke angiogenesis and metastasis. Targeting the exosomal EPHA2-AMPK signaling may serve as a potential strategy for breast cancer therapy.

Novel Anti-Cancer Products Targeting AMPK: Natural Herbal Medicine against Breast Cancer

“…AMPK is initiated to suppress Akt with the help of pleckstrin homology domain leucine-rich repeat protein phosphatases 2 (PHLPP2) to combat anoikis [ 55 ]. As an important part of cancer cell metastasis, AMPK activation can be induced by EPH receptor A2 (EPHA2)-enriched exosomes via a canonical forward signaling pathway that is dependent upon the ligand Ephrin A1, indicating that EPHA2, Ephrin A1, and HIF-1α, which are specifically contained in the tumor microenvironment, jointly promote AMPK activation and angiogenesis [ 118 ]. In addition, strong AMPK activation has been found to promote the detachment and anoikis resistance of MDA-MB-231 cells [ 119 ].…”

Perspective Chapter: Clinical Application of Exosome Components

“…Among them, exosomal glucose transporter 1 (GLUT-1), glypican 1 (GPC-1), and a disintegrin and metalloproteinase 10 (ADAM10) may be served as BC potential biomarkers [23]. Exosomes carrying proteins, including Anx II, Wnt7a, and ephrin type-A receptor 2 (EPHA2) could stimulate BC's invasive, angiogenesis, and metastasis abilities [24][25][26]. Exosomal Anx II has stimulated angiogenesis and BC metastasis [24].…”

“…Further, exosomal Wnt7a has been found to promote lung metastasis of BC [25]. Exosomal EPHA2 has also been found to activate the AMP-activated protein kinase (AMPK) signal pathway via the Ephrin A1-EPHA2 forward signal that promoted the angiogenesis and metastasis of BC [26]. Therefore, these exosome proteins may serve as potential BC therapeutic targets.…”

Exosomes in Mastitis—Research Status, Opportunities, and Challenges

“…The extensiveness of exosome sources and the richness of exosome contents determine their functions, which are diverse. Tumor cell-derived exosomes are widely involved in the occurrence and development of cancer [ 44 ], and affect tumor microenvironment remodeling [ 45 ], angiogenesis [ 46 ], tumor cell migration and invasion, and epithelial-mesenchymal transition (EMT) by mediating intercellular communication [ 47 ], drug resistance [ 48 , 49 ], and resistance to cell death [ 50 ]. In neurodegenerative diseases such as Alzheimer’s disease (AD) and Parkinson’s disease (PD), miRNAs in exosomes derived from peripheral blood and cerebrospinal fluid have shown promise as preclinical diagnostic markers [ 51 , 52 , 53 , 54 ].…”