title: Identifying differentially expressed genes in trophozoites and cysts of Acanthamoeba T4 genotype: Implications for developing new treatments for Acanthamoeba keratitis.
abstract: Acanthamoeba T4 genotype is the most prevalent genotype associated with amoebic keratitis. Acanthamoeba keratitis therapy is difficult due to transformation of trophozoite to cyst stage, which hinders the treatment of the disease. Although encystation assists the organism to survive against the chemotherapeutic compounds, the precise mechanism of encystation remains poorly understood. The purpose of this work was to identify differentially expressed genes in Acanthamoeba T4 genotype which might be useful for understanding of the encystment process and may thus help develop more efficient treatment. The mRNA profile of trophozoite and cyst of Acanthamoeba T4 genotype isolated from a soft contact lens wearer were analyzed using a cDNA ampliﬁed fragment length polymorphism (cDNA-AFLP) technique. Subsequently, a real time reverse transcriptase-PCR was performed to validate the cDNA-AFLP results. Three genes, heat shock protein70 (hsp70), actin-I and elongation factor-1alpha (EF-1α) were differentially expressed during Acanthamoeba differentiation. An in silico result predicted that transformation of trophozoite to cyst could be mediated through their cooperation with the protein partners interaction. Taken together, our experimental and bioinformatics findings suggested potential functions of hsp70, EF-1α and actin-I in differentiation of Acanthamoeba T4 genotype which may be useful in the design of an efficient therapeutic strategy in AK.
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Hoda
authors.initials: H
authors.lastname: Abedkhojasteh
authors.affiliation: Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Niyyati
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Sasan
authors.initials: S
authors.lastname: Rezaei
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Mohebali
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Shohreh
authors.initials: S
authors.lastname: Farnia
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Elham
authors.initials: E
authors.lastname: Kazemi-Rad
authors.affiliation: Stem Cell Preparation Unit, Farabi Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Roozafzoon
authors.affiliation: School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
authors.forename: Hamed
authors.initials: H
authors.lastname: Sianati
authors.affiliation: Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: rezaiian@tums.ac.ir.
authors.forename: Mostafa
authors.initials: M
authors.lastname: Rezaeian
authors.affiliation: Stem Cell Preparation Unit, Farabi Eye Research Center, Farabi Eye Hospital, Tehran University of Medical Sciences, Tehran, Iran; Department of Medical Genetics, Tehran University of Medical Sciences, Tehran, Iran. Electronic address: mheidari@sina.tums.ac.ir.
authors.forename: Mansour
authors.initials: M
authors.lastname: Heidari
mesh_terms: D000048:Acanthamoeba / Q000235:genetics* / Q000254:growth & development; D015823:Acanthamoeba Keratitis / Q000469:parasitology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017125:Genes, Protozoan / Q000235:genetics; D005838:Genotype; D053058:Trophozoites
keywords: Acanthamoeba keratitis; Genes modulating trophozoite-cyst transformation; In silico; Real-time RT-PCR; cDNA-AFLP