Circulating blood extracellular vesicles as a tool to assess endothelial injury and chemotherapy toxicity in adjuvant cancer patients

Abstract

Extracellular vesicles (EVs) are subcellular membrane blebs that include exosomes and microparticles, which represent a potential source for cancer biomarker discovery. We assess EVs characteristics as a tool to evaluate the endothelial and anti-tumor treatment injury during adjuvant chemotherapy in breast (BC) and colon cancer (CC) patients. Blood samples were taken from 29 BC and 25 CC patients before and after chemotherapy, as well as from healthy control donors (HC). Circulating blood EVs were isolated and characterized by size/concentration, membrane antigens for cell origin, thrombogenicity, and protein content. We observed higher EVs concentration and particle size in CC patients after chemotherapy compared with HC. Higher levels of endothelial EVs (CD144-positive) and vascular endothelial growth factor receptor 1 (VEGFR1), apparently as an indication of endothelial dysfunction, were found in all cancer patients, regardless of a given treatment, compared to HC. Levels of EVs labeled CD62E, CD34 + 41 - , the lymphocyte markers CD11 + and CD-14 + , Annexin-V, and the coagulation proteins TF and TFPI, however, sometimes demonstrate significant differences between patients, although HC did not show significant differences between patients pre- and post-chemotherapy. Most importantly, increasing levels of EVs encapsulated Angiostatin were found in patients with CC, while chemotherapy treatment leads to its notable rise in circulating blood EVs. Our results demonstrate the potential of EVs encapsulated Angiostatin as a tool to evaluate endothelial damage during adjuvant chemotherapy in BC and CC patients.

Extracellular Vesicles Released after Doxorubicin Treatment in Rats Protect Cardiomyocytes from Oxidative Damage and Induce Pro-Inflammatory Gene Expression in Macrophages

“…In contrast, exosomes from bone marrow- and adipose-derived mesenchymal stem cells protected DIC by inhibiting inflammatory responses and fibrosis [ 23 ]. EVs in circulation are heterogeneous in their tissue-cellular origin and have been proposed as a potential biomarker for DOXO-induced cardiomyopathy (DIC) [ 20 , 24 ]. However, their exact effects on normal cells after the cessation of DOXO treatment has never been clearly elucidated.…”

Differences in the Quantity and Composition of Extracellular Vesicles in the Aqueous Humor of Patients with Retinal Neovascular Diseases

“…They are composed of a lipid bilayer membrane and originate from the cellular endosome or plasma membrane and carry diverse cargo including specific proteins, RNA, DNA, and lipids depending on the secreted cell type [ 3 ]. Through their role of transporting and exchanging cargo between cells, EVs have been shown to be important mediators of intercellular communication in the maintenance of normal and pathophysiological conditions, such as cancer, neurodegenerative disorders, and infectious diseases [ 26 , 27 , 28 ]. Although several studies have shown their participation in the development of RPE cells and in murine models associated with eye diseases such as uveitis and AMD [ 25 ], the existence of EVs in patients suffering from other eye diseases remains unclear.…”

Isolation and Characterization of Extracellular Vesicles in Human Bowel Lavage Fluid

“…Moreover, the NTA results demonstrate that these EVs presented a regular size, considering that exosomes have a diameter of 30–100 nm and microvesicles have a diameter of 100 nm −1 µm [ 27 ]. The size difference between TEM and NTA results can be attributed to the dehydrating conditions to which EVs are subjected during fixation for TEM analysis [ 8 ], in addition to the presence of a sufficient concentration of EVs in BLF, taking into account that in urine, the EV concentration determined by NTA is 1.00 × 10 10 [ 28 ], whereas in CRC patient blood, the EV concentration is 1.29 × 10 9 ± 9.92 × 10 8 [ 29 ]. Finally, CD9 and CD63 expression, which are two tetraspanins considered EV biomarkers [ 8 ], has been found in EVs isolated from BLF.…”

“…EV cargo can be determined and studied, since RNA can be used to study gene expression levels [ 30 ], DNA mutations can be determined [ 31 ], and protein expression levels can be analyzed by Western blot and liquid chromatography–tandem mass spectrometry [ 29 , 32 , 33 , 34 ]. However, given the nature of our sample, it was necessary to confirm the content of these EVs from BLF as a feasible tool for CRC study.…”