Abstract
ABSTRACTExtracellular vesicles (EVs), including exosomes and microvesicles, are secreted from all cells, and convey messages between cells in health and disease. However, the diversity of EV subpopulations is only beginning to be explored. Since EVs have been implicated in tumour microenvironmental communication, we started to determine the diversity of EVs specifically in this tissue. To do this, we isolated EVs directly from patient melanoma metastatic tissues. Using EV membrane isolation and mass spectrometry analysis, we discovered enrichment of mitochondrial membrane proteins in the melanoma tissue-derived EVs, compared to non-melanoma-derived EVs. Interestingly, two mitochondrial inner membrane proteins MT-CO2 (encoded by the mitochondrial genome) and COX6c (encoded by the nuclear genome) were highly prevalent in the plasma of melanoma patients, as well as in ovarian and breast cancer patients. Furthermore, this subpopulation of EVs contains active mitochondrial enzymes. In summary, tumour tissues are enriched in EVs with mitochondrial membrane proteins and these mitochondrial membrane proteins can be detected in plasma and are increased in melanoma, ovarian cancer as well as breast cancer.
Highlights
Extracellular vesicles (EVs) are nano-sized (50–1000 nm in diameter) vesicles with a lipid bilayer membrane that play a significant role in mediating intercellular communication [1,2]
The existence of EVs in the interstitial space of a melanoma metastatic tissue was visualized by electron microscopy of tumour tissue pieces (Figure 1(a))
Melanoma metastatic tissue-derived EVs were isolated using an ultracentrifugation-based protocol that is able to separate EV subpopulations; vesicles are isolated at lower centrifugation speed (16,500 × g) and at higher speed (118,000 × g)
Summary
Extracellular vesicles (EVs) are nano-sized (50–1000 nm in diameter) vesicles with a lipid bilayer membrane that play a significant role in mediating intercellular communication [1,2]. EVs can be found in all body fluids, including blood [6], urine [7], ejaculate [8], and breast milk [9], and they are considered to carry signatures of the cells that produce them. This means that EVs have promising potential as diagnostic markers in disease, especially for cancers. We determined whether or not any such subpopulation is present in plasma from healthy individual vs patients with melanoma, ovarian cancer as well as breast cancer
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