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Introduction: Canine lymphoma (cL) is one of the most frequent cancers in dogs. The T-cell lymphoma (TcL) is not the most common phenotype but presents an aggressive behavior. MicroRNAs (miRNAs), are small, single-stranded, noncoding RNA molecules which can circulate freely in blood or be associated with extracellular vesicles (EVs). The dysregulation of certain miRNAs has been identified in numerous types of human cancers and they have been largely investigated as possible tumors biomarkers in human medicine, while research in veterinary oncology is still scarce. The aim of this study was to compare the expression patterns of free circulating and EV-associated miRNAs in dogs with T-cell lymhoma (TcL) and healthy dogs. Methods: Eight dogs with TcL were selected as the lymphoma group (LG) and eight dogs were included as controls (Ctrl). Plasma samples were collected at the time of the diagnosis and EVs isolated with ultracentrifugation. miRNAs were extracted from both the circulating EVs and the plasma supernatant, obtaining EV-associated and free-miRNAs. Quantitative real-time PCR was performed to analyze the expression of 88 target miRNAs. Results: Ten and seven differentially expressed miRNAs between LG and Ctrl were detected in EV-associated and free-miRNAs, respectively. Among EV-associated and free-miRNAs, only has-miR-222-3p was overexpressed in both conditions. Discussion: All the differentially expressed miRNAs detected in this study, have been already described as dysregulated in other human or canine cancers. The EV-associated miRNAs, which appear to be more stable and better conserved than free-miRNAs, could be investigated in further larger studies to better assess their use as possible biomarkers for TcL. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) are cell‐derived membrane‐bound vesicles involved in many biological processes such as tumour progression. For years, ultracentrifugation (UC) has been considered the gold standard for EV isolation but limited purity and integrity allowed the diffusion of alternative techniques. In this study, EVs were isolated from a canine mammary tumour cell line using UC and size exclusion chromatography (SEC) and analysed for size and concentration by nanoparticle tracking analysis (NTA) and for protein expression by western blot (WB). EV autocrine effect on cell proliferation, migration and invasiveness was then evaluated in vitro. In all samples, particles were in the EV size range (50–1000 nm), with a higher concentration in UC than in SEC samples (1011 and 1010 particles/ml respectively), and expressed EV markers (Alix, CD9). Functional assays did not show statistically significant difference among conditions, but EV treatment slightly increased cell proliferation and invasiveness and treatment with SEC‐isolated EVs slightly enhanced cell migration compared to UC‐isolated EVs. In conclusion, the main differences between the two isolation techniques are the quantity of the final EV‐product and slight differences on EV functionality, which should be further explored to better highlight the real autocrine effect of tumoral EVs. [ABSTRACT FROM AUTHOR]

Extracellular vesicles (EVs) are double-layered lipid membrane vesicles released by cells. Currently, EVs are attracting a lot of attention in the biological and medical fields due to their role as natural carriers of proteins, lipids, and nucleic acids. Thus, they can transport useful genomic information from their parental cell through body fluids, promoting cell-to-cell communication even between different organs. Due to their functionality as cargo carriers and their protein expression, they can play an important role as possible diagnostic and prognostic biomarkers in various types of diseases, e.g., cancers, neurodegenerative, and autoimmune diseases. Today, given the invaluable importance of EVs, there are some pivotal challenges to overcome in terms of their isolation. Conventional methods have some limitations: they are influenced by the starting sample, might present low throughput and low purity, and sometimes a lack of reproducibility, being operator dependent. During the past few years, several microfluidic approaches have been proposed to address these issues. In this review, we summarize the most important microfluidic-based devices for EV isolation, highlighting their advantages and disadvantages compared to existing technology, as well as the current state of the art from the perspective of the use of these devices in clinical applications. [ABSTRACT FROM AUTHOR]

Advances in tumour research are crucial, and comparative oncology can improve the knowledge in several ways. Dogs are not only models of specific naturally occurring tumours but can also be sentinels of environmental exposures to carcinogens, as they share the same environment with their owners. The purpose of this work was to describe the data collected by The Italian Network of Laboratories for Veterinary Oncology in the first 9 years of activity (2013-2021) and to evaluate their potential epidemiological significance. Frequencies of tumour topographies and main morphologies in dogs were described, analysed and compared, calculating age-adjusted proportional morbidity ratios and considering several risk factors (breed, sex, period and region of residence). These observations allowed us to highlight differences not only in morphology and topography of some tumours but also to formulate hypotheses on the potential role of some risk factors, e.g., neutering/spaying or geographical location. In our opinion, the results of this case series confirm the importance of initiating and consolidating animal cancer registration initiatives that would facilitate the possibility of conducting multicentric collaborative studies to deepen the knowledge of the epidemiology of tumours in dogs from a comparative perspective.