Theranostics 2022; 12(9):4127-4146. doi:10.7150/thno.72404 This issue

Research Paper

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

Baoai Han1,2,3,4, He Zhang1,2,3,4, Ruinan Tian1,2,3,4, Hui Liu1,2,3,4, Zhaosong Wang1,2,3,4, Zhiyong Wang1,2,3,4, Jianfei Tian1,2,3,4, Yanfen Cui1,2,3,4, Sixin Ren1,2,3,4, Xiaoyan Zuo1,2,3,4, Ran Tian1,2,3,4, Ruifang Niu1,2,3,4,✉, Fei Zhang1,2,3,4,✉

1. Public Laboratory, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, 300060, China.
2. Key Laboratory of Cancer Prevention and Therapy, Tianjin, 300060, China.
3. Tianjin's Clinical Research Center for Cancer, Tianjin, 300060, China.
4. Key Laboratory of Breast Cancer Prevention and Therapy, Ministry of Education, Tianjin, 300060, China.

This is an open access article distributed under the terms of the Creative Commons Attribution License ( See for full terms and conditions.
Han B, Zhang H, Tian R, Liu H, Wang Z, Wang Z, Tian J, Cui Y, Ren S, Zuo X, Tian R, Niu R, Zhang F. Exosomal EPHA2 derived from highly metastatic breast cancer cells promotes angiogenesis by activating the AMPK signaling pathway through Ephrin A1-EPHA2 forward signaling. Theranostics 2022; 12(9):4127-4146. doi:10.7150/thno.72404. Available from

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Graphic 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.

Keywords: exosomes, angiogenesis, high metastatic potential cells, breast cancer, EPHA2