title: Autophagy inhibitors as a potential antiamoebic treatment for Acanthamoeba keratitis.
abstract: Acanthamoeba cysts are resistant to extreme physical and chemical conditions. Autophagy is an essential pathway for encystation of Acanthamoeba cells. To evaluate the possibility of an autophagic Acanthamoeba encystation mechanism, we evaluated autophagy inhibitors, such as 3-methyladenine (3MA), LY294002, wortmannin, bafilomycin A, and chloroquine. Among these autophagy inhibitors, the use of 3MA and chloroquine showed a significant reduction in the encystation ratio in Acanthamoeba cells. Wortmannin also inhibited the formation of mature cysts, while LY294002 and bafilomycin A did not affect the encystation of Acanthamoeba cells. Transmission electron microscopy revealed that 3MA and wortmannin inhibited autophagy formation and that chloroquine interfered with the formation of autolysosomes. Inhibition of autophagy or autolysosome formation resulted in a significant block in the encystation in Acanthamoeba cells. Clinical treatment with 0.02% polyhexamethylene biguanide (PHMB) showed high cytopathic effects on Acanthamoeba trophozoites and cysts; however, it also revealed high cytopathic effects on human corneal epithelial cells. In this study, we investigated effects of the combination of a low (0.00125%) concentration of PHMB with each of the autophagy inhibitors 3MA, wortmannin, and chloroquine on Acanthamoeba and human corneal epithelial cells. These new combination treatments showed low cytopathic effects on human corneal cells and high cytopathic effects on Acanthamoeba cells. Taken together, these results provide fundamental information for optimizing the treatment of Acanthamoeba keratitis.
authors.affiliation: Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Taegu, Republic of Korea.
authors.forename: Eun-Kyung
authors.initials: EK
authors.lastname: Moon
authors.affiliation: Department of Parasitology, Dong-A University College of Medicine, Busan, Republic of Korea.
authors.forename: So-Hee
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Taegu, Republic of Korea.
authors.forename: Yeonchul
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Taegu, Republic of Korea.
authors.forename: Dong-Il
authors.initials: DI
authors.lastname: Chung
authors.affiliation: Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Taegu, Republic of Korea.
authors.forename: Youn-Kyoung
authors.initials: YK
authors.lastname: Goo
authors.affiliation: Department of Parasitology, Dong-A University College of Medicine, Busan, Republic of Korea hhkong@dau.ac.kr.
authors.forename: Hyun-Hee
authors.initials: HH
authors.lastname: Kong
mesh_terms: D000048:Acanthamoeba / Q000187:drug effects* / Q000648:ultrastructure; D015823:Acanthamoeba Keratitis / Q000188:drug therapy* / Q000469:parasitology; D000818:Animals; D000981:Antiprotozoal Agents / Q000627:therapeutic use*; D001343:Autophagy / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D003315:Cornea / Q000166:cytology; D004847:Epithelial Cells / Q000187:drug effects / Q000469:parasitology; D006801:Humans; D007634:Keratitis / Q000188:drug therapy* / Q000469:parasitology; D008247:Lysosomes / Q000187:drug effects; D053058:Trophozoites / Q000187:drug effects
chemical_list: D000981:Antiprotozoal Agents