Extracellular RNA profile of Echinococcus multilocularis metacestodes with different tegument integrity

ANCAROLA, MARÍA EUGENIA; LICHTENSTEIN, GABRIEL; HERBIG, JOHANNES; HOLROYD, NANCY; BERRIMAN, MATTHEW; ALBRECHT, KRYSTYNA; MARCILLA, ANTONIO; ROSENZVIT, MARA CECILIA; KAMENETZKY, LAURA; BREHM, KLAUS; CUCHER, MARCELA

Congreso; XXXII Reunión Anual de la Sociedad Argentina de Protozoología; 2020

Sociedad Argentina de Protozoología

The cestode parasite Echinococcus multilocularis is the causative agent of alveolar echinococcosis, characterized by growth of the metacestode stage in host liver, forming a heterogeneous multivesicular mass of peripheral proliferative tissue with central necrotic regions. The metacestode is a fluid-filled vesicle with an outer acellular laminated layer and an inner cellular germinal layer. This disease lacks biomarkers for early and/or follow-up diagnosis and thus detection of parasite extracellular RNA (ex-RNA) could be a potential novel diagnostic tool. Ex-RNA can be secreted in extracellular vesicles (EV), associated to lipoproteins or in ribonucleoprotein complexes. We have previously reported that two cestode species secrete ex-RNA in EV and that E. multilocularis produce EV that are retained by the laminated layer, hindering their release to the extra-parasite milieu.In this work, we have identified the ex-RNA profile secreted by E. multilocularis metacestodes to the extra- and intra-parasite milieus in cultures classified as active or transitional according to tegument integrity.For this, we analysed active (>95% vital) and transitional (70-85% vital) metacestode cultures and by differential centrifugation we isolated EV-enriched (P100) and EV-depleted (S100) fractions of culture medium and metacestode fluid. Then, by nanoparticle tracking analysis we observed a significantly higher amount of EV in transitional cultures in both the extra- and intra-parasite P100 fractions. Then, we identified the ex-RNA profile by RNA sequencing and observed small RNAs