Presentation Information
[P03-430]Pro-angiogenic activity of cancer cell-derived migrasomes evaluated using a peptide-modified substrate
○Yuka Iwasaki1, Shogo Saito1, Mina Okochi1 (1. Institute of Science Tokyo (Japan))
Keywords:
Migrasomes,Extracellular vesicles,Angiogenesis,Peptide
[Background and purpose]
Migrasomes, extracellular vesicles formed on retraction fibers during cell migration, have attracted attention as carriers that dynamically and spatially regulate intercellular communication. However, functional analysis of migrasomes has been limited by the lack of suitable isolation methods that preserve their native properties. In the tumor microenvironment, angiogenesis is a critical process that supports tumor growth and metastasis, and is regulated by cancer cell–derived soluble factors and extracellular vesicles. Nevertheless, the contribution of migrasomes to angiogenesis remains largely unclear. In this study, we investigated the angiogenesis-promoting activity of cancer cell–derived migrasomes and whether this activity depends on the malignancy of donor cells.
[Methods]
To preserve the intrinsic properties of migrasomes, we developed a novel isolation method using a peptide-modified substrate. Specifically, cells were cultured on substrates functionalized with an integrin-binding RGD peptide and a cell-penetrating pVEC peptide. Following EDTA treatment to detach cells, migrasomes were captured and recovered directly on the substrate. Migrasomes were collected from highly malignant breast cancer cells (MDA-MB-231) and non-tumorigenic mammary epithelial cells (MCF-10A). Human umbilical vein endothelial cells (HUVECs) were seeded onto migrasomes-captured substrates, and their angiogenic responses were evaluated by tube formation assays on extracellular matrix (ECM) gels and gene expression analysis of the angiogenesis-related factor cyclooxygenase-2 (COX-2).
[Results and Discussion]
HUVECs treated with cancer cell–derived migrasomes exhibited markedly enhanced tube formation, including increased network complexity and branching structures, compared with untreated controls and those treated with migrasomes derived from non-tumorigenic cells. In addition, COX-2 expression in HUVECs increased over time following uptake of cancer cell–derived migrasomes, suggesting activation of pro-angiogenic signaling pathways in a malignancy-dependent manner. These effects were consistent with those observed using migrasomes isolated by conventional ultracentrifugation, supporting the validity of the peptide-modified substrate–based method. Notably, this platform enables functional evaluation of migrasomes under conditions that better preserve their key structural features and spatial context.These findings demonstrate that cancer cell–derived migrasomes enhance the angiogenic potential of endothelial cells in a malignancy-dependent manner. Furthermore, our results suggest that migrasomes function as extracellular mediators that facilitate localized communication between cancer cells and endothelial cells within the tumor microenvironment. The peptide-modified substrate presented here provides a novel analytical platform for studying intact migrasomes and their biological functions. This study highlights a previously underappreciated role of migrasomes in tumor-associated angiogenesis and provides a foundation for further investigation of their underlying molecular mechanisms and potential therapeutic applications.
Migrasomes, extracellular vesicles formed on retraction fibers during cell migration, have attracted attention as carriers that dynamically and spatially regulate intercellular communication. However, functional analysis of migrasomes has been limited by the lack of suitable isolation methods that preserve their native properties. In the tumor microenvironment, angiogenesis is a critical process that supports tumor growth and metastasis, and is regulated by cancer cell–derived soluble factors and extracellular vesicles. Nevertheless, the contribution of migrasomes to angiogenesis remains largely unclear. In this study, we investigated the angiogenesis-promoting activity of cancer cell–derived migrasomes and whether this activity depends on the malignancy of donor cells.
[Methods]
To preserve the intrinsic properties of migrasomes, we developed a novel isolation method using a peptide-modified substrate. Specifically, cells were cultured on substrates functionalized with an integrin-binding RGD peptide and a cell-penetrating pVEC peptide. Following EDTA treatment to detach cells, migrasomes were captured and recovered directly on the substrate. Migrasomes were collected from highly malignant breast cancer cells (MDA-MB-231) and non-tumorigenic mammary epithelial cells (MCF-10A). Human umbilical vein endothelial cells (HUVECs) were seeded onto migrasomes-captured substrates, and their angiogenic responses were evaluated by tube formation assays on extracellular matrix (ECM) gels and gene expression analysis of the angiogenesis-related factor cyclooxygenase-2 (COX-2).
[Results and Discussion]
HUVECs treated with cancer cell–derived migrasomes exhibited markedly enhanced tube formation, including increased network complexity and branching structures, compared with untreated controls and those treated with migrasomes derived from non-tumorigenic cells. In addition, COX-2 expression in HUVECs increased over time following uptake of cancer cell–derived migrasomes, suggesting activation of pro-angiogenic signaling pathways in a malignancy-dependent manner. These effects were consistent with those observed using migrasomes isolated by conventional ultracentrifugation, supporting the validity of the peptide-modified substrate–based method. Notably, this platform enables functional evaluation of migrasomes under conditions that better preserve their key structural features and spatial context.These findings demonstrate that cancer cell–derived migrasomes enhance the angiogenic potential of endothelial cells in a malignancy-dependent manner. Furthermore, our results suggest that migrasomes function as extracellular mediators that facilitate localized communication between cancer cells and endothelial cells within the tumor microenvironment. The peptide-modified substrate presented here provides a novel analytical platform for studying intact migrasomes and their biological functions. This study highlights a previously underappreciated role of migrasomes in tumor-associated angiogenesis and provides a foundation for further investigation of their underlying molecular mechanisms and potential therapeutic applications.
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