Expression profiling of Echinococcus multilocularis miRNAs throughout metacestode development in vitro

MACCHIAROLI, N.; PREZA, M.; PEREZ,M.; KAMENETZKY, L.; CUCHER, M.; MALDONADO, L.; KOZIOL, U.; CASTILLO, E.; BREHM, K.; ROSENZVIT, M.

PLOS NEGLECTED TROPICAL DISEASES

PUBLIC LIBRARY SCIENCE

Lugar: San Francisco; Año: 2021

1935-2735

The neglected zoonotic disease alveolar echinococcosis (AE) is caused bythe metacestode stage of the helminth parasite Echinococcus multilocularis. Current chemotherapeutical treatmentrequires prolonged drug theraphy using benzimidazoles that are parasitostaticonly. Thus, novel strategies for the treatment of AE are urgently needed.MicroRNAs (miRNAs) are a class of small non-coding RNAs with a major role inregulation of gene expression in key biological processes. In previous work, weanalyzed the miRNA expression profile of E.multilocularisin vivo metacestodes and invitro primary cell cultures. However, current knowledge about miRNAexpression and their potential rolesin E.multilocularis developmentis still limited. In this work, we analyzed forthe first known time the expression profile of E. multilocularismiRNAs throughout metacestode development in vitro, determined thespatial expression of miR-71 in in vitro metacestodesand predicted miRNAfunctions. Small RNAlibraries from different samples of E.multilocularis metacestodes, primary cell cultures and protoescoleces weresequenced. After miRNA identification, expression analyses and targetprediction were performed. We identified a repertoire of 37 miRNAs in E.multilocularis,some of them absent in the host or highlydivergent from their hostorthologs.MiRNA expressionprofiling revealed a small number of highly expressed miRNAs in all samplesanalyzed. The high expression of these miRNAs wasconserved in other cestodes,suggesting essential roles in development, survival or host-parasite interaction.Interestingly, some of these highly expressed miRNAs were absent or divergentfrom human host, suggestingtheir potential use as therapeutic targets. MiRNAdifferential expression analyses showed highly regulated miRNAs during thedifferent transitions analyzed, suggesting a role in the regulation ofdevelopmental timing, host-parasite interaction, and/or in maintaining the unique developmental features of E. multilocularis metacestode. We determinedthat miR-71 is expressed in stemcells and in other cell types of the germinal layer in E. multilocularis in vitrometacestodes. Functional analyses of predicted miRNAtargets reveals conserved functions for highly expressed miRNAs and essential roles in parasite biology.The evolutionaryconservation and expressionanalyses of E.multilocularis miRNAs throughout the metacestode development along with thefunctional analyses of their predicted targets might help to identify selectivetherapeutic targets for treatment and control of alveolar echinococcosis.