Liquid biopsy based on small extracellular vesicles predicts chemotherapy response of canine multicentric lymphomas

Taismara K Garnica,Juliano C Silveira,Ana C F C M Ávila,Aline Zoppa,Oscar R S Matiz,Arina L Rochetti,Roana C S Ribeiro,Ricardo F Strefezzi,Jéssika C C Lesbon,Mirela T Costa,Adriana T Nishiya,David J Argyle,Heidge Fukumasu,Andrigo B De Nardi

doi:10.1038/s41598-020-77366-7

Abstract

Lymphoma is the most common type of canine hematological malignancy where the multicentric (cMCL) form accounts for 75% of all cases. The standard treatment is the CHOP chemotherapy protocols that include cyclophosphamide, doxorubicin, vincristine and prednisone, where the majority of dogs achieve complete/partial response; however, it is very important to predict non-responsive cases to improve treatment and to develop new targeted therapies. Here we evaluate a liquid biopsy approach based on serum Small Extracellular Vesicles enriched for exosomes (SEVs) to predict cMCL chemotherapy response. Nineteen dogs at the end of the 19-week chemotherapy protocol (8 Complete Response and 11 Progressive Disease) were evaluated for serum SEVs size, concentration and screened for 95 oncomirs. PD patients had higher SEVs concentration at the diagnosis than CR patients (P = 0.034). The ROC curve was significant for SEVs concentration to predict the response to CHOP (AUC = 0.8011, P = 0.0287). A potential molecular signature based on oncomirs from SEVs (caf-miR-205, caf-miR-222, caf-mir-20a and caf-miR-93) is proposed. To the best of our knowledge, this is the first study demonstrating the potential of a liquid biopsy based on SEVs and their miRNAs content to predict the outcome of chemotherapy for canine multicentric lymphomas.

Highlights

Lymphoma is the most common type of canine hematological malignancy where the multicentric form accounts for 75% of all cases
The chemoresistance and relapse are events commonly observed during the treatment of canine lymphomas being directly related to therapeutic efficacy and survival[29]
Data from the literature show that 85% of dogs with lymphoma treated with CHOP 19 weeks protocol, achieves complete or partial response[30]

Summary

Introduction

Lymphoma is the most common type of canine hematological malignancy where the multicentric (cMCL) form accounts for 75% of all cases. To the best of our knowledge, this is the first study demonstrating the potential of a liquid biopsy based on SEVs and their miRNAs content to predict the outcome of chemotherapy for canine multicentric lymphomas. Exosomes are small extracellular vesicle (30–150 nm) with double lipidic membrane and can be involved in cellular communication as well as transporting of important biological molecules (mRNA, miRNA, metabolites, proteins, receptors) between c ells[16] These vesicles are enrolled in cancer development and can be detect in organic fluids such as blood, urine, s aliva[17,18], eliciting exosomes as potential candidates for liquid biopsy approaches[19]. An in vitro study showed three exosomal miRNAs (miR-151, miR-8908a-3p, and miR-486) and CD82 protein with different expression between vincristine-sensitive canine cancer cell lines (CLBL-1 and GL-1) and the resistant cell line (UL-1)[24]

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