Expression of Mir-21 and Mir-103a in Toxocara canis: Potential for Diagnosis of Human Toxocariasis
-
Vahid RAISSI
,
Mehdi MOHEBALI
,
Eshrat Bigom KIA
,
Abbas RAHIMI FOROUSHANI
,
Nasrin SOHRABI
,
Mohammad Bagher ROKNI
,
Mohammad ZIBAEI
Abstract
Background: Toxocariasis is one of the most neglected zoonotic diseases, predominantly caused by Toxocara canis. We aimed to evaluate the expression of microRNAs 21 and 103a in seropositive individuals for human toxocariasis as diagnostic biomarkers.
Methods: This study was conducted on 324 individuals for ELISA test on toxocariasis in Tehran and Karaj, Iran 2019. Then positive samples for anti-Toxocara IgG were obtained to quantitative Real-time PCR (qRT-PCR) assays to investigate the transcriptional profiles of miRNAs predicted to be involved in developmental and reproductive processes. qPCR was employed to assess levels of transcription for miRNAs of 103a and 21 in plasma samples.
Results: After the experiments, the results were evaluated by REST software, Livak formula and quantitative t-test. The analyzes performed on human samples showed that in the case group compared to the control group, only in Tc-miR-21 gene, a 0.3-fold increase in expression was obtained with REST software (Fold change ≤ 1.5, P>0.05), which was statistically significant by t-test (P<0.05).
Conclusion: To our knowledge, this is the first study to evaluate miR-21 and miR-103a in toxocariasis, which shed light on the fundamental role of it as a biomarker and diagnostic tool. However, due to the changes in expression of these miRNAs were not vast to be used as biomarkers in diagnosis. Despite of that the changes in the expression of these miRNAs were not vast but they could serve as novel promising biomarkers for diagnosis of toxocariasis.
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25. Britton C, Winter AD, Marks ND, et al. Application of small RNA technology for improved control of parasitic helminths. Vet Parasitol. 2015;212(1-2):47-53.
26. Li Z, Chen X, Zen X, et al. MicroRNA expression profile in the third- and fourth-stage larvae of Angi-ostrongylus cantonensis. Parasitol Res. 2014;113(5):1883-1896.
27. Sun J, Wang S, Li C, et al. Novel expression profiles of microRNAs suggest that specific miRNAs regu-late gene expression for the sexual maturation of female Schistosoma japonicum after pairing. Parasit Vec-tors. 2014;7:177.
28. Zhao GH, Xu MJ, Zhu XQ. Identification and characterization of microRNAs in Baylisascaris schroed-eri of the giant panda. Parasit Vectors. 2013;6:216.
29. Ahmed R, Chang Z, Younis AE, et al. Conserved miRNAs are candidate post-transcriptional regula-tors of developmental arrest in free-living and para-sitic nematodes. Genome Biol Evol. 2013;5(7):1246-1260.
30. Devaney E, Winter AD, Britton C. microRNAs: a role in drug resistance in parasitic nematodes?. Trends Parasitol. 2010;26(9):428-433.
31. Winter AD, Weir W, Hunt M, et al. Diversity in parasitic nematode genomes: the microRNAs of Brugia pahangi and Haemonchus contortus are largely novel. BMC Genomics. 2012;13(1):4.
32. Poole CB, Davis PJ, Jin J, et al. Cloning and bioin-formatic identification of small RNAs in the filarial nematode, Brugia malayi. Mol Biochem Parasitol. 2010; 169(2):87-94.
33. Wang J, Czech B, Crunk A, et al. Deep small RNA sequencing from the nematode Ascaris reveals conservation, functional diversification, and novel developmental profiles. Genome Res. 2011;21(9):1462-1477.
34. Ma G, Luo Y, Zhu H, et al. MicroRNAs of Toxo-cara canis and their predicted functional roles. Parasit Vectors. 2016;9:229.
35. Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function. Nucleic Acids Res. 2019;47(D1): D155-D162.
36. Zibaei M, Sadjjadi SM, Maraghi S. The occurrence of Toxocara species in naturally infected broiler chickens revealed by molecular approaches. J Hel-minthol. 2017;91(5):633-636.
2. Raissi V, Sohrabi Z, Getso M, et al. Risk factors and prevalence of toxocariasis in pregnant women and diabetic patients compared to healthy adults in Ilam province, western Iran. EXCLI J. 2018;17:983-988.
3. Raissi V, Taghipour A, Navi Z, et al. Seropreva-lence of Toxoplasma gondii and Toxocara spp. infec-tions among pregnant women with and without previous abortions in the west of Iran. J Obstet Gynaecol Res. 2020;46(3):382-388.
4. Zibaei M, Sadjjadi SM, Uga S. Experimental Toxo-cara cati infection in gerbils and rats. Korean J Parasi-tol. 2010;48(4):331-333.
5. Rostami A, Riahi SM, Holland CV, et al. Seropreva-lence estimates for toxocariasis in people world-wide: A systematic review and meta-analysis. PLoS Negl Trop Dis. 2019;13(12):e0007809.
6. Zibaei M, Sadjjadi SM, Sarkari B, et al. Evaluation of Toxocara cati Excretory-Secretory Larval Antigens in Serodiagnosis of Human Toxocariasis. J Clin Lab Anal. 2016;30(3):248-253.
7. Chen J, Liu Q, Liu G, et al. Toxocariasis: a silent threat with a progressive public health impact. In-fect Dis Poverty. 2018;7(1):59.
8. Gasser RB. A perfect time to harness advanced molecular technologies to explore the fundamental biology of Toxocara species. Vet Parasitol. 2013;193(4):353-364.
9. Zhu XQ, Korhonen PK, Cai H, et al. Genetic blueprint of the zoonotic pathogen Toxocara canis. Nat Commun. 2015;6:6145.
10. Gu HY, Marks ND, Winter AD, et al. Conserva-tion of a microRNA cluster in parasitic nematodes and profiling of miRNAs in excretory-secretory products and microvesicles of Haemonchus contortus. PLOS Negl Trop Dis. 2017;11(11):e0006056.
11. Manzano-Román R, Siles-Lucas M. MicroRNAs in parasitic diseases: potential for diagnosis and target-ing. Mol Biochem Parasitol. 2012;186(2):81-86.
12. Hakimi MA, Cannella D. Apicomplexan parasites and subversion of the host cell microRNA pathway. Trends Parasitol. 2011;27(11):481-486.
13. Wang K, Yuan Y, Cho J-H, et al. Comparing the MicroRNA spectrum between serum and plasma. PloS One. 2012;7(7):e41561.
14. Tsui NBY, Ng EKO, Lo YMD. Stability of en-dogenous and added RNA in blood specimens, se-rum, and plasma. Clin Chem. 2002;48(10):1647-1653.
15. Gevaert AB, Witvrouwen I, Vrints CJ, et al. MicroRNA profiling in plasma samples using qPCR arrays: Recommendations for correct analysis and interpretation. PloS One. 2018;13(2):e0193173.
16. Wang L, Wang HC, Chen C, et al. Differential expression of plasma miR-146a in sepsis patients compared with non-sepsis-SIRS patients. Exp Ther Med. 2013;5(4):1101-1104.
17. Pfaffl MW, Horgan GW, Dempfle L. Relative expression software tool (REST©) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res. 2002;30(9):e36.
18. Rao X, Huang X, Zhou Z, et al. An improvement of the 2ˆ (-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3(3):71-85.
19. Giorgi G, Marcantonio P, Del Re B. LINE-1 retrotransposition in human neuroblastoma cells is affected by oxidative stress. Cell Tissue Res. 2011; 346(3):383-391.
20. Chen X, Li ZY, Maleewong W, et al. Serum aca-mir-146a is a potential biomarker for early diagnosis of Angiostrongylus cantonensis infection. Parasitol Res. 2014; 113(9):3221-3227.
21. Nunes S, Silva IB, Ampuero MR, et al. Integrated analysis reveals that miR-193b, miR-671, and TREM-1 correlate with a good response to treat-ment of human localized cutaneous leishmaniasis caused by Leishmania braziliensis. Front Immunology. 2018;9:640.
22. Macha MA, Seshacharyulu P, Krishn SR, et al. MicroRNAs (miRNAs) as biomarker(s) for prog-nosis and diagnosis of gastrointestinal (GI) cancers. Curr Pharm Des. 2014;20(33):5287-5297.
23. MacLellan SA, MacAulay C, Lam S, et al. Pre-profiling factors influencing serum microRNA levels. BMC Clin Pathol. 2014;14:27.
24. Turchinovich A, Cho W. The origin, function and diagnostic potential of extracellular microRNA in human body fluids. Front Genet. 2014;5:30.
25. Britton C, Winter AD, Marks ND, et al. Application of small RNA technology for improved control of parasitic helminths. Vet Parasitol. 2015;212(1-2):47-53.
26. Li Z, Chen X, Zen X, et al. MicroRNA expression profile in the third- and fourth-stage larvae of Angi-ostrongylus cantonensis. Parasitol Res. 2014;113(5):1883-1896.
27. Sun J, Wang S, Li C, et al. Novel expression profiles of microRNAs suggest that specific miRNAs regu-late gene expression for the sexual maturation of female Schistosoma japonicum after pairing. Parasit Vec-tors. 2014;7:177.
28. Zhao GH, Xu MJ, Zhu XQ. Identification and characterization of microRNAs in Baylisascaris schroed-eri of the giant panda. Parasit Vectors. 2013;6:216.
29. Ahmed R, Chang Z, Younis AE, et al. Conserved miRNAs are candidate post-transcriptional regula-tors of developmental arrest in free-living and para-sitic nematodes. Genome Biol Evol. 2013;5(7):1246-1260.
30. Devaney E, Winter AD, Britton C. microRNAs: a role in drug resistance in parasitic nematodes?. Trends Parasitol. 2010;26(9):428-433.
31. Winter AD, Weir W, Hunt M, et al. Diversity in parasitic nematode genomes: the microRNAs of Brugia pahangi and Haemonchus contortus are largely novel. BMC Genomics. 2012;13(1):4.
32. Poole CB, Davis PJ, Jin J, et al. Cloning and bioin-formatic identification of small RNAs in the filarial nematode, Brugia malayi. Mol Biochem Parasitol. 2010; 169(2):87-94.
33. Wang J, Czech B, Crunk A, et al. Deep small RNA sequencing from the nematode Ascaris reveals conservation, functional diversification, and novel developmental profiles. Genome Res. 2011;21(9):1462-1477.
34. Ma G, Luo Y, Zhu H, et al. MicroRNAs of Toxo-cara canis and their predicted functional roles. Parasit Vectors. 2016;9:229.
35. Kozomara A, Birgaoanu M, Griffiths-Jones S. miRBase: from microRNA sequences to function. Nucleic Acids Res. 2019;47(D1): D155-D162.
36. Zibaei M, Sadjjadi SM, Maraghi S. The occurrence of Toxocara species in naturally infected broiler chickens revealed by molecular approaches. J Hel-minthol. 2017;91(5):633-636.
| Files | ||
| Issue | Vol 15 No 4 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v15i4.4865 | |
| Keywords | ||
| Toxocariasis miRNAs expression Polymerase chain reac-tion | ||
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How to Cite
1.
RAISSI V, MOHEBALI M, KIA EB, RAHIMI FOROUSHANI A, SOHRABI N, ROKNI MB, ZIBAEI M. Expression of Mir-21 and Mir-103a in Toxocara canis: Potential for Diagnosis of Human Toxocariasis. Iran J Parasitol. 2020;15(4):559-567.
