Identification and expression profiling of microRNAs in Hymenolepis

Macchiaroli, Natalia; Cucher, Marcela Alejandra; Kamenetzky, Laura; Yones, Cristian Ariel; Bugnon, Leandro A.; Berriman, Matt; Olson, Peter D.; Rosenzvit, Mara Cecília

doi:10.1016/j.ijpara.2018.07.005

Cited by 16 publications

(26 citation statements)

References 81 publications

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“…Small RNA isolation, library construction and sequencing of three biological replicates from 48 h primary cell cultures of E. multilocularis (Fig 1) were carried out as described in Macchiaroli et al [23]. Briefly, cells were mechanically homogenized in Trizol (Invitrogen) for 10 s. Then, 200 μl of chloroform:isoamyl alcohol (24:1) were added and mixed thoroughly.…”

Section: Small Rna Isolation Small Rna Library Construction and Sequmentioning

confidence: 99%

Deciphering the role of miR-71 in Echinococcus multilocularis early development in vitro

et al. 2019

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Echinococcosis represents a major public health problem worldwide and is considered a neglected disease by the World Health Organization. The etiological agents are Echinococcus tapeworms, which display elaborate developmental traits that imply a complex control of gene expression. MicroRNAs (miRNAs), a class of small regulatory RNAs, are involved in the regulation of many biological processes such as development and metabolism. They act through the repression of messenger RNAs (mRNAs) usually by binding to the 3' untranslated region (3'UTR). Previously, we described the miRNome of several Echinococcus species and found that miRNAs are highly expressed in all life cycle stages, suggesting an important role in gene expression regulation. However, studying the role of miRNAs in helminth biology remains a challenge. To develop methodology for functional analysis of miR-NAs in tapeworms, we performed miRNA knockdown experiments in primary cell cultures of Echinococcus multilocularis, which mimic the development of metacestode vesicles from parasite stem cells in vitro. First, we analysed the miRNA repertoire of E. multilocularis primary cells by small RNA-seq and found that miR-71, a bilaterian miRNA absent in vertebrate hosts, is one of the top five most expressed miRNAs. Using genomic information and bioinformatic algorithms for miRNA binding prediction, we found a high number of potential miR-71 targets in E. multilocularis. Inhibition of miRNAs can be achieved by transfection of antisense oligonucleotides (anti-miRs) that block miRNA function. To this end, we evaluated a variety of chemically modified anti-miRs for miR-71 knockdown. Electroporation of primary cells with 2'-O-methyl modified anti-miR-71 led to significantly reduced miR-71 levels. Transcriptomic analyses showed that several predicted miR-71 targets were up-regulated in anti-miR-treated primary cells, including genes potentially involved in parasite development, host parasite interaction, and several genes of as yet unknown function. Notably, miR-71silenced primary cell cultures showed a strikingly different phenotype from control cells and did not develop into fully mature metacestodes. These findings indicate an important function of miR-71 in Echinococcus development and provide, for the first time, methodology to functionally study miRNAs in a tapeworm.

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Section: Small Rna Isolation Small Rna Library Construction and Sequmentioning

confidence: 99%

Deciphering the role of miR-71 in Echinococcus multilocularis early development in vitro

et al. 2019

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“…The existing studies on miRNAs in cestodes have mainly focused on miRNA predictions of target genes, which provided a platform for the identification of miRNAs using both computational and experimental approaches. However, there is still a lack of comprehensive functional and experimental validation of these putative miRNA targets [20]. Studying the role of miRNAs in parasite immunity still remains a challenge [21].…”

Section: Introductionmentioning

confidence: 99%

MiR-374b-5p Regulates T Cell Differentiation and Is Associated with rEg.P29 Immunity

Zhu

et al. 2020

BioMed Research International

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Cystic echinococcosis (CE) is a zoonotic disease caused by Echinococcus granulosus (Eg) infection. Our previous study confirmed that recombinant Eg.P29 (rEg.P29) could protect against echinococcus granulosus secondary infection in sheep and mice. The aim of the study was to investigate the association between immunoprotection of rEg.P29 vaccine and mmu-miR-374b-5p (miR-374b-5p) and study the immunity influence of miR-374b-5p on CD4+ T cells in mice spleen. MiR-374b-5p level was significantly increased after the second-week and the fourth week of vaccination with rEg.P29. Overexpression of miR-374b-5p increased IFN-γ, IL-2, IL-17A mRNA levels and decreased IL-10 mRNA levels in CD4+ T cells. Moreover, the inhibition of miR-374b-5p decreased IFN-γ and IL-17A and increased IL-10 mRNA levels in CD4+ T cells; this was further confirmed by the flow cytometry. The vaccination of rEg.P29 enhanced miR-374b-5p expression that was associated with a higher Th1 and Th17 immune response, a lower IL-10 mRNA production with miR-374b-5p overexpression, a lower Th1 immune response, and a higher IL-10 mRNA levels with miR-374b-5p inhibitions. To sum up, these data suggest that miR-374b-5p may participate in rEg.P29 immunity by regulating Th1 and Th17 differentiation.

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“…Once the discovery of microRNAs, a small noncoding RNAs that act as endogenous silencers of numerous target genes [7], their remarkable role in the regulation of wide range of signaling pathways has been identified [8]. Although some microRNAs serve as oncogenes and activate cancer-related signaling pathways, the others could participate as tumor suppressors, affecting the expression of various biomolecules, such as neuropilin-1, metalloproteinase 14 (MMP-14), Snail and so forth that are important for the formation of the malignancies [9][10][11][12]. Moreover, the unique profile of microRNA expression in human cancer cells and their tissue-specific manner made these small RNAs a potent biomolecules for both cancer diagnosis and screening [13].…”

Section: Introductionmentioning

confidence: 99%

Evaluating the expression level of miR-9-5p and miR-192-5p in gastrointestinal cancer: introducing novel screening biomarkers for patients

2020

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Objective: It has been indicated that there is a tight association between cancer and different factors, such as environment and genetics, including aberrantly expressed microRNAs. The crucial role of microRNAs in the regulation of diverse signaling pathways in gastrointestinal cancer has been established in several studies. In this study, we aimed to evaluate the expression of microRNA-9 and -192 in colon and gastric cancers. After extracting the RNA from tissues and serum samples of patients, suffering from colon and gastric cancer, cDNA was synthesized. Then by performing quantitative real-time PCR, we evaluated the expression level of miR-9-5p and miR-192-5p in collected samples.Results: Unlike to colon cancer in which the expression level of miR-9-5p remained unchanged, the relative expression of this miRNA decreased remarkably in gastric cancer (with P value < 0.05), in comparison with normal adjacent tissues. In agreement with this finding, we also found that the expression level of miR-192-5p was decreased in gastric cancer tissues, compared to normal gastric tissue. Given the reduction in the expression level of miR-9-5p and miR-192-5p in gastric cancer, it could be postulated to consider these miRNAs as promising diagnostic biomarkers.

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Identification and expression profiling of microRNAs in Hymenolepis

Cited by 16 publications

References 81 publications

Deciphering the role of miR-71 in Echinococcus multilocularis early development in vitro

Deciphering the role of miR-71 in Echinococcus multilocularis early development in vitro

MiR-374b-5p Regulates T Cell Differentiation and Is Associated with rEg.P29 Immunity

Evaluating the expression level of miR-9-5p and miR-192-5p in gastrointestinal cancer: introducing novel screening biomarkers for patients

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