MicroRNAs Expression Induces Apoptosis of Macrophages in Response to Leishmania major (MRHO/IR/75/ER): An In-Vitro and In-Vivo Study

  • Mostafa GHOLAMREZAEI Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Soheila ROUHANI ORCID Mail Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Mehdi MOHEBALI ORCID Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran AND Center for Research of Endemic Parasites of Iran (CREPI), Tehran University of Medical Sciences, Tehran, Iran
  • Samira MOHAMMADI-YEGANEH ORCID Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran AND Department of Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Mostafa HAJI MOLLA HOSEINI ORCID Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Ali HAGHIGHI ORCID Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Zohreh LASJERDI Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Faezeh HAMIDI Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  • Mohammad KAZEM SHARIFI-YAZDI Zoonosis Research Center, Tehran University of Medical Sciences, Tehran, Iran
Keywords:
Leishmania major, Apoptosis, microRNAs, In vitro, In vivo

Abstract

Background: We aimed to investigate the effect of miR-15a mimic and inhibitor of miR-155 expression on apoptosis induction in macrophages infected with Iranian strain of Leishmania major in-vitro and in-vivo.

Methods: RAW 264.7 cells were infected with L. major promastigotes (MRHO/IR/75/ER), and then were treated with miRNAs. For in-vivo experiment, BALB/c mice were inoculated with L. major promastigotes, and then they were treated with miRNAs. For evaluation of miRNA therapeutic effect, in-vitro and in-vivo studies were performed using quantitative Real-time PCR, Flow cytometry, lesion size measurement, and Limiting Dilution Assay (LDA). This study was performed in Shahid Beheshti University of Medical Sciences in 2019.

Results: In-vitro results of flow cytometry showed that using miR-15a mimic, miR-155 inhibitor or both of them increased apoptosis of macrophages. In in-vivo, size of lesion increased during experiment in control groups (P<0.05) while application of both miR-155 inhibitor and miR-15a mimic inhibited the increase in the size of lesions within 6 wk of experiment (P=0.85). LDA results showed that microRNA therapy could significantly decrease parasite load in mimic or inhibitor receiving groups compared to the control group (P<0.05).

Conclusion: miR-155 inhibitor and miR-15a mimic in L. major infected macrophages can induce apoptosis and reduce parasite burden. Therefore, miRNA-based therapy can be proposed as new treatment for cutaneous leishmaniasis.

References

1. Ghatee MA, Walter RT, Karamian M. The geo-graphical distribution of cutaneous leishmaniasis causative agents in Iran and its neighboring coun-tries, A Review. Front Public Health. 2020; 8: 11.
2. Bailey MS, Lockwood DNJ. Cutaneous leishmania-sis. Clin Dermatol. 2007; 25: 203–211.
3. Holakouie-Naieni K, Mostafavi E, Darvishi Boloo-rani A, et al. Spatial modeling of cutaneous leish-maniasis in Iran from 1983 to 2013. Acta Trop. 2017; 166: 67-73.
4. Gholamrezaei M, Mohebali M, Hanafi-Bojd AA, et al. Ecological niche modeling of main reservoir hosts of zoonotic cutaneous leishmaniasis in Iran. Acta Trop. 2016; 160: 44-52.
5. Duclos S, Desjardins M. Subversion of a young phagosome: the survival strategies of intracellular pathogens. Cell Microbiol. 2000; 2(5):365-77.
6. Frank B, Marcu A, Luis de Oliveira Almeida Pe-tersen A, et al. Autophagic digestion of Leishmania major by host macrophages is associated with differ-ential expression of BNIP3, CTSE, and the miR-NAs miR-101c, miR-129, and miR-210. Parasit Vectors. 2015; 8:404.
7. Liu D, Uzonna J E. The early interaction of Leish-mania with macrophages and dendritic cells and its influence on the host immune response. Front Cell Infect Microbiol. 2012; 2:83.
8. Antoine J C, Prina E, Lang Th, et al. The biogenesis and properties of the parasite- ophorous vacuoles that harbor Leishmania in murine macrophages. Trends Microbiol. 1998; 6(10): 392-401.
9. Mohebali M, Kazemirad E, Hajjaran H, et al. Gene expression analysis of antimony resistance in Leish-mania tropica using quantitative real-time PCR fo-cused on genes involved in trypanothione metabo-lism and drug transport. Arch Dermatol Res. 2019; 311(1): 9-17.
10. Mohtasebi S, Mohebali M, Elikaee S, et al. In vitro and in vivo anti-parasitic activity of biogenic antimo-ny sulfide nanoparticles on Leishmania major (MRHO/IR/75/ER). Parasitol Res. 2019; 118(9): 2669-78.
11. Salehi Sangani Gh, Jajarmi V, Khamesipour A, et al. Generation of a CRISPR/Cas9-Based Vector Spe-cific for Gene Manipulation in Leishmania major. Iran J Parasitol. 2019; 14(1): 78-88.
12. Aghaei M, Khanahmad H, Aghaei S, et al. Evolu-tion of transgenic L. infantum expressing mLLO-BAX-SMAC in the infected macrophages apopto-sis in vitro and in vivo. Parasite Immunol. 2020; 42(11):e12726.
13. Vaux DL, Strasser A. The molecular biology of apoptosis. Proc Natl Acad Sci U S A. 1996; 93(6): 2239-44.
14. Igney FH, Krammer PH. Death and anti-death: tumour resistance to apoptosis. Nat Rev Cancer. 2002; 2(4): 277-88.
15. Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004; 116(2): 281-97.
16. Cai Y, Yu X, Hu S, et al. A Brief Review on the Mechanisms of miRNA Regulation. Genomics Proteomics Bioinformatics. 2009; 7(4): 147-54.
17. Abdullah OA, El Gazzar WB, Salem TI, et al. miR-15a: A Potential Diagnostic Biomarker and a Candi-date for Non-operative Therapeutic Modality for Age-related Cataract. Br J Biomed Sci. 2019; 76(4): 184-189.
18. Zhang H, Li Y, Huang Q, et al. MiR-148a pro-motes apoptosis by targeting Bcl-2 in colorectal cancer. Cell Death Differ. 2011; 18(11): 1702–10.
19. Alizadeh Sh, Kaviani S, Soleimani M, et al. Mir-55 inhibition can reduce cell proliferation and induce apoptosis in Jurkat (Acute T cell Leukemia) cell line. Iran J Ped Hematol Oncol. 2014; 4(4): 141-50.
20. De Santis R, Liepelt A, Mossanen J C, et al. miR-155 targets Caspase-3 mRNA in activated macro-phages. RNA Biol .2016;13(1): 43–58.
21. Kropf P, Kadolsky UD, Rogers M, et al. The Leishmaniasis Model. Methods in Microbiology. 2010; 37: 307–28.
22. Hadighi R, Mohebali M, Boucher P, et al. Unre-sponsiveness to Glucantime treatment in Iranian cutaneous leishmaniasis due to drug-resistant Leish-mania tropica parasites. PLoS Med. 2006; 3(5): e162.
23. Laskay T, van Zandbergen G, Solbach W. Neutro-phil granulocytes – Trojan horses for Leishmania ma-jor and other intracellular microbes?. Trends Micro-biol. 2003; 11(5): 210-4.
24. Manzano-Román R, Siles-Lucas M. MicroRNAs in parasitic diseases: potential for diagnosis and target-ing. Mol Biochem Parasitol. 2012; 186(2): 81-6.
25. Liu Q, Tuo W, Gao H, et al. MicroRNAs of para-sites: current status and future perspectives. Parasi-tol Res. 2010; 107(3): 501–07.
26. Sahoo GC, Ansari MY, Dikhit MR, et al. Computa-tional identification of microRNA-like elements in Leishmania major. Microrna. 2014; 2(3): 225-30.
27. Moore KJ, Matlashewsk G. Intracellular infection by Leishmania donovani inhibits macrophage apopto-sis. J Immunol. 1994; 152(6):2930-37.
28. Kaufmann SH, Hengartner MO. Programmed cell death: alive and well in the new millennium. Trends Cell Biol. 2001; 11(12): 526-34.
29. Paris C, Loiseau PM, Bories Ch, et al. Miltefosine Induces apoptosis-like death in Leishmania donovani promastigotes. Antimicrob Agents Chemother. 2004; 48(3): 852-9.
30. Esmaeili J, Mohebali M, Edrissian Gh, et al. Evalua-tion of miltefosine against Leishmania major (MRHO/IR/75/ER): in vitro and in vivo studies. Acta Medica Iranica. 2008; 46(3): 191- 6.
31. Jha TK, Sundar S, Thakur CP, et al. Miltefosine, an oral agent, for the treatment of Indian visceral leishmaniasis. N Engl J Med. 1999; 341(24): 1795-800.
32. Resende TAC, Bernardes VF, Silva JC, et al. Loss of heterozygosity of MIR15A/MIR16-1, negative regulators of the antiapoptotic gene BCL2, is not common in odontogenic keratocysts. Oral Surg Oral Med Oral Pathol Oral Radiol. 2018; 125(4): 313-6.
33. Sicard F, Gayral M, Lulka H, et al. Targeting miR-21for the therapy of pancreatic cancer. Mol Ther. 2013; 21(5): 986-94.
34. Lasjerdi Z, Ghanbarian H, Mohammadi Yeganeh S, et al. The comparative expression profile analysis of apoptosis-related miRNA and its target gene in Leishmania major–infected macrophages. Iran J Para-sitol. 2020;15(3):332-340.
35. Hashemi N, Sharifi MR, Masjedi M, et al. Locked nucleic acid -anti- let-7a induces apoptosis and ne-crosis in macrophages infected with Leishmania ma-jor. Microb Pathog. 2018; 119: 193-99.
Published
2020-12-06
How to Cite
1.
GHOLAMREZAEI M, ROUHANI S, MOHEBALI M, MOHAMMADI-YEGANEH S, HAJI MOLLA HOSEINI M, HAGHIGHI A, LASJERDI Z, HAMIDI F, KAZEM SHARIFI-YAZDI M. MicroRNAs Expression Induces Apoptosis of Macrophages in Response to Leishmania major (MRHO/IR/75/ER): An In-Vitro and In-Vivo Study. Iran J Parasitol. 15(4):475-487.
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