Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Isolates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran

  • Yasaman Alijani Research Center for Food Safety and Health, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran AND 2Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
  • Saeedeh Sadat Hosseini Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
  • Salman Ahmadian Research Center for Food Safety and Health, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran AND Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
  • Sonia Boughattas Biomedical Research Center, Qatar University, Doha, Qatar
  • Gilda Eslami Research Center for Food Safety and Health, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran AND Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
  • Shadi Naderian Department of Statistics and Epidemiology, School of Public Health, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
  • Vahid Ajamian Research Center for Food Safety and Health, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran AND Department of Parasitology and Mycology, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran
Keywords: Leishmaniasis, Cutaneous, Drug rsesistance, Antimony

Abstract

Background: Regarding the antimonial-resistant of Leishmania spp., understanding of related mechanism is neces­sary. One of the most important involved molecules is aquaglyceropin1 (AQP1). The aim of this study was molecu­lar analysis of AQP1 gene from antimonial-resistant clinical isolates and its expression.Methods: Overall, 150 patients with cutaneous leishmaniasis referring to the reference laboratories of Yazd and Varzaneh,, located 105km southeast of Isfahan and 240km away from Yazd, were assessed from Jun 2015 to Dec 2017. After sampling, staining was done and evaluated for Leishman by microscope. Samples were collected in RNAlater solution for gene expression analysis in non-healing isolates. DNA extraction was performed from each slide with Leishman body. All patients with L. major isolates detected by ITS1-PCR-RFLP were followed for find­ing the resistant isolates, consequence of molecular characterization of AQP1 using PCR-RFLP. Gene expression of AQP1 from all resistant isolates was assessed in comparison with the one in a sensitive isolate. Statistical analysis was done using SPSS. The significance level was considered ≤0.05.Results: Five isolates were detected as antimonial resistant. Molecular detection and identification were appeared that all were L. major. The molecular characterization of AQP1 showed G562A mutation. Gene expression of AQP1 in resistant isolates showed 1.67 fold higher than the sensitive isolate.Conclusion: We reported a new point mutation of G562A in AQP1 gene involved in molecular mechanism in re­sistant isolates.

References

Eslami G, Hajimohammadi B, Jafari AA, Mirzaei F, Gholamrezai M, Anvari H, Khamesipour A (2014) Molecular identification of Leishmania tropica infections in patients with cutaneous leishmaniasis from an endemic central of Iran. Trop Biomed. 31(4): 592–599.

Eslami G, Zarchi MV, Moradi A, Hejazi SH, Sohrevardi SM, Vakili M, Khamesi-pour A (2016) Aquaglyceroporin1 gene expression in antimony resistance and susceptible Leishmania major isolates. J Vector Borne Dis. 53(4): 370–374.

Naouel E, Ihcene KD, Sofiane B, Khati-ma AO, Razika B, Bruno O, Zoubir H, Denis S (2017) Antimonial suscepti-bility and in vivo behavior of Leishmania major isolates collected in Al-geria before and after treatment. Acta Trop. 180: 7–11.

Hajjaran H, Kazemi-Rad E, Mohebali M, Oshaghi MA, Khadem-Erfan MB, Hajaliloo E, Reisi Nafchi H, Raoofian R (2016) Expression analysis of activated protein kinase C gene (LACK1) in antimony sensitive and resistant Leishmania tropica clinical isolates using real-time RT-PCR. Int J Derma-tol. 55(9): 1020–1026.

Plourde M, Ubeda JM, Mandal G, Mon-te-Neto RL, Mukhopadhyay R, Ouellette M (2015) Generation of an aquaglyceroporin AQP1 null mutant in Leishmania major. Mol Biochem Parasitol. 201(2): 108–111.

Kazemi-Rad E, Mohebali M, Khadem-Erfan MB, Saffari M, Raoofian R, Hajjaran H, Hadighi R, Khamesipour A, Rezaie S, Abedkhojasteh H, Heidari M (2013) Identification of antimony resistance markers in Leishmania tropica field isolates through a cDNA-AFLP ap-proach. Exp Parasitol. 135(2): 344–349.

Monte-Neto R, Laffitte MC, Leprohon P, Reis P, Frézard F, Ouellette M (2015) Intrachromosomal amplification, locus deletion and point mutation in the aquaglyceroporin AQP1 gene in anti-mony resistant Leishmania (Viannia) guyanensis. PLoS Negl Trop Dis. 9(2): e0003476.

Mukherjee A, Boisvert S, Monte-Neto RL, Coelho AC, Raymond F, Mukho¬padhyay R, Corbeil J, Ouellette M (2013) Telomeric gene deletion and intrachromo-somal amplification in antimony-resistant Leishmania. Mol Microbiol. 88(1): 189–202.

Leprohon P, Fernandez-Prada C, Gazanion É, Monte-Neto R, Ouellette M (2014) Drug resistance analysis by next generation sequencing in Leishmania. Int J Parasitol Drugs Drug Resist. 5(1): 26–35.

Eslami G, Salehi R, Khosravi S, Doudi M (2012) Genetic analysis of clinical isolates of Leishmania major from Is-fahan, Iran. J Vector Borne Dis. 49(3): 168–174.

Maharjan M, Singh S, Chatterjee M, Madhubala R (2008) Role of aquaglyceroporin (AQP1) gene and drug uptake in antimony-resistant clinical isolates of Leishmania donovani. Am J Trop Med Hyg. 79(1): 69–75.

Figarella K, Uzcategui NL, Zhou Y, LeFurgey A, Ouellette M, Bhattacharjee H, Mukhopadhyay R (2007) Biochemical characterization of Leishmania major aquaglyceroporin LmAQP1: possible role in volume regulation and osmotaxis. Mol Microbiol. 65(4): 1006–1017.

Haile S, Papadopoulou B (2007) Devel-opmental regulation of gene expres-sion in trypanosomatid parasitic pro-to¬zoa. Curr Opin Microbiol. 10(6): 569–577

Mandal G, Sharma M, Kruse M, Sander-Juelch C, Munro LA, Wang Y, Vilg JV, Tamás MJ, Bhattacharjee H, Wiese M, Mukhopadhyay R (2012) Modulation of Leishmania major aquaglyceroporin activity by a mitogenactivated protein kinase. Mol Microbiol. 85(6): 1204–1218.

Jeddi F, Mary C, Aoun K, Harrat Z, Bouratbine A, Faraut F, Benikhlef R, Pomares C, Pratlong F, Marty P, Piarroux R (2014) Heterogeneity of moecular resistance patterns in antimony-resistant field isolates of Leishma-nia species from the western Mediterranean area. Antimicrob Agents Chemother. 58 (8): 4866–4874.

Mandal G, Sarkar A, Saha P, Singh N, Sundar S, Chatterjee M (2009) Functionality of drug efflux pumps in antimonial resistant Leishmania donovani field isolates. Indian J Biochem Biophys. 46(1): 86–92.

Ashutosh, Sundar S, Goyal N (2007) Molecular mechanisms of antimony resistance in Leishmania. J Med Microbiol. 56(Pt 2): 143–153.

Sharma M, Mandal G, Mandal S, Bhattacharjee H, Mukhopadhyay R (2015) Functional role of lysine 12 in Leishmania major AQP1. Mol Biochem Parasitol. 201(2): 139–145.

Mukhopadhyay R, Mandal G, Atluri VS, Figarella K, Uzcategui NL, Zhou Y, Beitz E, Ajees AA, Bhattacharjee H (2011) The role of alanine 163 in so-lute permeability of Leishmania major aquaglyceroporin LmAQP1. Mol Biochem Parasitol. 175(1): 83–90

Uzcategui NL, Zhou Y, Figarella K, Ye J, Mukhopadhyay R, Bhattacharjee H (2008) Alteration in glycerol and metalloid permeability by a single mutation in the extracellular C-loop of Leishmania major aquaglyceroporin LmAQP1. Mol Microbiol. 70(6): 1477–1486.

Published
2019-05-25
How to Cite
1.
Alijani Y, Hosseini SS, Ahmadian S, Boughattas S, Eslami G, Naderian S, Ajamian V. Molecular Analysis of Aquaglyceroporin 1 Gene in Non-Healing Clinical Isolates Obtained from Patients with Cutaneous Leishmaniasis from Central of Iran. J Arthropod Borne Dis. 13(2):145-152.
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Original Article