TGF-β Targeted by miR-27a Modulates Anti-Parasite Responses of Immune System

Faezeh  Hamidi; Samira  Mohammadi-Yeganeh; Mostafa Haji Molla  Hoseini; Seyyed Javad  Seyyed Tabaei; Niloofar  Taghipour; Ameneh  Koochaki; Vahedeh  Hosseini; Ali  Haghighi

doi:10.18502/ijpa.v18i3.13762

Faezeh Hamidi Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Samira Mohammadi-Yeganeh Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Mostafa Haji Molla Hoseini Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Seyyed Javad Seyyed Tabaei Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Niloofar Taghipour Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Ameneh Koochaki Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Vahedeh Hosseini Department of Molecular Medicine and Genetics, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
Ali Haghighi Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

DOI: https://doi.org/10.18502/ijpa.v18i3.13762

Keywords: Leishmania major; Luciferase assay; Immunity

Abstract

Background: Immune cells and their secreted cytokines are known as the first barrier against pathogens. Leishmania major as an intracellular protozoan produces anti-inflammatory cytokines that lead to proliferation and survival of the parasite in the macrophages. miRNAs are small non-coding RNA molecules that regulate mRNAs expression. We aimed to investigate the relationship between the expression of TGF-β and a bioinformatically candidate miRNA, in leishmaniasis as a model of TGF-β overexpression.

Methods: The miRNAs that target TGF-β -3´UTR were predicted and scored by bioinformatic tools. After cloning of TGF-β-3'UTR in psi-CHECK ^TM- 2 vector, targeting validation was confirmed using Luciferase assay. After miRNA mimic transfection, the expression of miR-27a, TGF-β, as well as Nitric Oxide concentration was evaluated.

Results: miR-27a received the highest score for targeting TGF-β in bioinformatic predictions. Luciferase assay confirmed that miR-27a is targeting TGF-β-3'UTR, since miR-27a transfection decreased the luciferase activity. After miRNA transfection, TGF-β expression and Nitric Oxide concentration were declined in L. major infected macrophages.

Conclusion: Bioinformatic prediction, luciferase assay, and miRNA transfection results showed that miR-27a targets TGF-β. Since miRNA and cytokine-base therapies are developing in infectious diseases, finding and validating miRNAs targeting regulatory cytokines can be a novel strategy for controlling and treating leishmaniasis.