A comparative analysis of extracellular vesicles (EVs) from human and feline plasma

Abstract

Extracellular vesicles (EVs) are nanoparticles found in all biological fluids, capable of transporting biological material around the body. Extensive research into the physiological role of EVs has led to the development of the Minimal Information for Studies of Extracellular Vesicles (MISEV) framework in 2018. This framework guides the standardisation of protocols in the EV field. To date, the focus has been on EVs of human origin. As comparative medicine progresses, there has been a drive to study similarities between diseases in humans and animals. To successfully research EVs in felines, we must validate the application of the MISEV guidelines in this group. EVs were isolated from the plasma of healthy humans and felines. EV characterisation was carried out according to the MISEV guidelines. Human and feline plasma showed a similar concentration of EVs, comparable expression of known EV markers and analogous particle to protein ratios. Mass spectrometry analyses showed that the proteomic signature of EVs from humans and felines were similar. Asymmetrical flow field flow fractionation, showed two distinct subpopulations of EVs isolated from human plasma, whereas only one subpopulation was isolated from feline plasma. Metabolomic profiling showed similar profiles for humans and felines. In conclusion, isolation, and characterisation of EVs from humans and felines show that MISEV2018 guidelines may also be applied to felines. Potential comparative medicine studies of EVs may provide a model for studying naturally occurring diseases in both humans and felines.

Acute phase proteins in cats: Diagnostic and prognostic role, future directions, and analytical challenges

“…Although limited due to incompletely characterized genome sequences and incomplete annotation of gene functions in domestic species, 65 over the last decade, proteomics have been incorporated in a variety of research studies in cats 120–124 . Proteomic analyses of different biological fluids in healthy and diseased cats have been recently performed, setting a new trend in proteomic research 31,125–127 . Nevertheless, there is still a paucity of studies investigating inflammatory markers through proteomics.…”

“…[120][121][122][123][124] Proteomic analyses of different biological fluids in healthy and diseased cats have been recently performed, setting a new trend in proteomic research. 31,[125][126][127] Nevertheless, there is still a paucity of studies investigating inflammatory markers through proteomics. To the author's knowledge, only one study investigated potential inflammatory markers in healthy cats and those with pancreatitis or pancreatic carcinoma.…”

The separation and identification of circulating small extracellular vesicles from endurance‐trained, strength‐trained and recreationally active men

“…Although the presence of phosphatidylserine in EV membranes is well established as essential for the recognition and uptake of EVs into recipient cells (Buzás et al, 2018), the relevance of free serine for EV biogenesis and/or EV function is not immediately clear. However, serine has previously been detected in preparations of small EVs derived from human and feline plasma (Howard et al, 2022), human serum (Luo et al, 2018) and human urine (Puhka et al, 2017). Serine is a substrate for enzymes such as serine palmitoyltransferase, an enzyme essential for the production of ceramide (Hanada, 2003).…”

“…Metabolomic analysis was performed using a SCIEX QTRAP 6500+ mass spectrometer coupled to SCIEX ExionLC Series with ultra‐high‐performance liquid chromatography capability. This approach has previously been employed in preparations of human‐derived circulating small EVs and is more extensively detailed elsewhere (Howard et al., 2022). Briefly, amino acids ( n = 21) and biogenic amines ( n = 21) were targeted in LC–MS/MS analysis using isotopically labelled internal standards and 7‐point calibration curves.…”

The protein and miRNA profile of plasma extracellular vesicles (EVs) can distinguish feline mammary adenocarcinoma patients from healthy feline controls

“…EVs were isolated from plasma by size exclusion chromatography (SEC), using IZON qEV original columns (iZon Science), according to previously published methods 30 . Briefly, the columns were removed from 4 °C to room temperature the 20% ethanol storage solution was removed, and the column flushed with 10 ml Phosphate Buffered Saline (PBS).…”