microRNAs in Taenia crassiceps and Taenia solium: characterization and future applications

MATIAS PEREZ; MARCELA CUCHER; NATALIA MACCHIAROLI; MARA ROSENZVIT

Congreso; Reunión Conjunta 2016 SAIC-SAI-SAFE; 2016

The cestode parasite Taenia solium is the etiologicalagent of neurocisticercosis, one of the 17 neglected tropicaldiseases prioritized by the WHO. Taenia crassiceps,another cestode from the same genus, is used as a laboratorymodel of T. solium. microRNAs (miRNAs) are smallnon-coding RNAs considered master regulators of geneexpression with key roles in diverse cellular processes.At present there is no evidence of miRNAs from theseparasites. High-throughput characterization of small RNAs(sRNAs) in T. Crassice􀁒s larval stage (cisticercus) ispresented. 􀀴􀀰A 􀁙as purified from cysticerci 􀀊􀀕 biologicalreplicates), sRNA library was constructed and sequencedwith HiSeq 2500. The mirDeep2 software was used formi􀀴􀀰A identification. The T. 􀀵olium genome was usedfor bioinformatics analyses, since it is the closest genomeavailable, allowing also to identify T. solium miRNAs atthe genome level. Experimental validation of selected sequenceswas performed by Northern blotting. The resultsobtained showed that miRNAs were the most abundantcategory of sRNAs accounting for 83% of mapped reads. Afinal high confidence set of 􀀖􀀔 mi􀀴􀀰As􀀜 􀀕􀀚 conserved and4 novel, was obtained. Northern blot results showed bandscompatible with miRNA biogenesis, allowing validatingidentified mi􀀴􀀰As. Expression analysis sho􀁙ed that fe􀁙miRNAs accounted for most miRNA expression. Previousresults of our group (Cucher et al, 2015; Macchiaroli et al,2015, Basika et al, 2016) showed that these set of miRNAsis conserved and also highly expressed in other cestodesincluding Echinococcus s􀁒􀁒, suggesting important rolesin cestode biology. We are currently predicting genesregulated by the highly expressed miRNAs in cestodes,some of which are absent or divergent in the mammalhosts. This is the first report of mi􀀴􀀰As in T.crassice􀁒sand T. solium. Highly expressed parasite miRNAs absentor divergent in the hosts 􀁙ere indentified and could becandidates for drug and diagnosis targeting.