| Identification of essential genes for Acanthamoeba castellanii excystation during encystation and excystation |
Min-Jeong Kim1,†
, Hye-Jeong Jo1,†
, Fu-Shi Quan2,3
, Ki Back Chu4,5
, Hyun-Hee Kong6
, Eun-Kyung Moon2
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1Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Korea; 2Department of Medical Zoology, Kyung Hee University School of Medicine, Seoul 02447, Korea
2Department of Medical Zoology, Kyung Hee University School of Medicine, Seoul 02447, Korea
3Medical Research Center for Bioreaction to Reactive Oxygen Species and Biomedical Science Institute, Core Research Institute, School of Medicine, Kyung Hee University, Seoul 02447, Korea
4Department of Parasitology, Inje University College of Medicine, Busan 47392, Korea
5Department of Infectious Disease and Malaria, Paik Institute of Clinical Research, Inje University, Busan 47392, Korea
6Department of Parasitology, Dong-A University College of Medicine, Busan 49201, Korea |
| † These authors contributed equally to this work. |
| * Corresponding Author:
Eun-Kyung Moon, Email: ekmoon@khu.ac.kr |
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Received: August 23, 2024; Accepted: September 30, 2024. |
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| Abstract |
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Acanthamoeba is an opportunistic pathogen that causes Acanthamoeba keratitis, granulomatous amoebic encephalitis, and other cutaneous diseases. The life cycle of Acanthamoeba consists of 2 stages of trophozoites and cysts. Under adverse environmental conditions, Acanthamoeba encysts, while the conditions become favorable for growth, it reverts to the trophozoite form. Acanthamoeba excystation is crucial for its proliferation and can lead to recurrent infections after incomplete treatment. To identify the factors involved in excystation, A. castellanii was subjected to either encystation- or excystation-inducing conditions, and gene expression profiles were compared using mRNA sequencing. A. castellanii samples were collected at 8 h intervals for analysis under both conditions. Differentially expressed gene analysis revealed that 1,214 and 1,163 genes were upregulated and downregulated, respectively, by more than 2-fold during early excystation. Five genes markedly upregulated in early excystation (ACA1_031140, ACA1_032330, ACA1_374400, ACA1_275740, and ACA1_112650) were selected, and their expression levels were confirmed via real-time PCR. Small interfering RNA (siRNA) targeting these 5 genes was transfected into Acanthamoeba and gene knockdown was validated through real-time PCR. The silencing of ACA1_031140, ACA1_032330, ACA1_374400, and ACA1_112650 inhibited excystation and suggested that these genes might be essential for excystation. Our findings provide valuable insights for suppressing Acanthamoeba proliferation and recurrence. |
| Key words:
Acanthamoeba, encystation, excystation, gene knockdown |
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