This project aims at developing new therapeutic tools to contribute to the control of neglected diseases caused by helminth parasites such as Echinococcus multilocularis, the aetiological agent of alveolar echinococcosis. Due to the scarcity of available anthelmintic drugs and the possible emergence of resistance, the discovery of new anthelmintic drugs is mandatory. MicroRNAs are small noncoding RNAs that have emerged as important regulators of gene expression and exert critical functions in diverse biological events. miRNA profiles in several cestodes have been discovered and subsequently characterised (Cucher et al, 2011; Macchiaroli et al., 2015; Cucher et al., 2015, Basika et al., 2016, Perez et al., 2017), miRNA-targeted genes and been bioinformatically predicted and their transport between cells also reported (Ancarola et al, 2017). miRNAs are the preponderant small RNA silencing molecules of several cestodes and found highly expressed parasite miRNAs, such as emu-miR-71 that are absent or highly divergent with the host homolog. Because of their essential roles, the development of chemical probes that selectively target a miRNA could be extraordinarily powerful. The general objective of this study is to determine the effect of miRNA silencing in E. multilocularis development as a first step to analyze the potential of miRNAs as drug targets, and to learn more about the regulation of primary cells in flatworms. Specifically, the aim is to compare by RNAseq the mRNA expression of miR-71 silenced vs control E. multilocularis primary cells. This will be done by in vitro miRNA silencing: introduction of antimiRs (oligonucleotides complementary to the miRNA that inhibit miRNA function) in vitro to cultured primary cells from E. multilocularis.
Alongside the search for miRNA targets using RNASeq, the project will evaluate the change in the level of the targeted miRNAs, alongside evaluation of a change in parasite viability, development and proliferation. A comparison of mRNA expression in miR-71 silenced vs control cultures will be undertaken. Those genes whose expression is higher in miR-71 silenced cultures with respect to control (mock electroporated and scramble electroporated) and that have been predicted to contain a miRNA recognition site (Macchiaroli et al, 2017) will be considered as potential miRNA targets. Expected outcome will be to understand miRNA role in E. multilocularis biology and evaluate their role in E. multilocularis development. This project will inform the wider aims of FUGI, the Wellcome Trust Strategic Award funded project exploring flatworm stem cell biology
This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
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