Gene editing of a carcinogenic liver fluke tetraspanin impairs parasite surface biogenesis and extracellular vesicle uptake by human host cells

Authors

Sujittra Chaiyadet, Department of Tropical Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002  Thailand.
Wannaporn Ittiprasert, Department of Microbiology, Immunology and Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, D.C. 20037, USA.
Michael J. Smout, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, 4878, Australia.
Ladawan Khowawisetsut, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Apisit Ruangsuwast, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002  Thailand.
Paul J. Brindley, Department of Microbiology, Immunology and Tropical Medicine, School of Medicine & Health Sciences, George Washington University, Washington, D.C. 20037, USA.
Alex Loukas, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, 4878, Australia.
Thewarach Laha, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002  Thailand.

Document Type

Journal Article

Publication Date

9-4-2025

Journal

The Journal of infectious diseases

DOI

10.1093/infdis/jiaf466

Keywords

Opisthorchis; CRISPR-Cas9; endocytosis; extracellular vesicle; liver fluke; tegument; tetraspanin

Abstract

Opisthorchiasis remains a significant public health concern throughout Southeast Asia. The liver fluke Opisthorchis viverrini resides within the biliary tract and chronic infection leads to bile duct cancer, or cholangiocarcinoma. Here, we examined the functions of liver fluke tetraspanins, four-transmembrane domain proteins expressed on the surface of the fluke tegument and extracellular vesicles (EVs) derived from this syncytial surface. We undertook CRISPR-Cas9 gene knockout (KO) of the O. viverrini tetraspanin-2 (Ov-tsp-2) gene and found that Ov-tsp-2-KO flukes had abnormal tegument biogenesis. The tegument of Ov-tsp-2-KO flukes was increasingly vacuolated and fewer EVs were secreted. EVs that were secreted were deficient in Ov-TSP-2 and their uptake by cholangiocytes was diminished. The findings indicate a critical role for Ov-TSP-2 in maintenance of the tegument, EV production and uptake by host target cells, and support the development of this parasite antigen as an anti-infection and anti-cancer vaccine for opisthorchiasis and opisthorchiasis-associated cholangiocarcinoma.

APA Citation

Chaiyadet, Sujittra; Ittiprasert, Wannaporn; Smout, Michael J.; Khowawisetsut, Ladawan; Ruangsuwast, Apisit; Brindley, Paul J.; Loukas, Alex; and Laha, Thewarach, "Gene editing of a carcinogenic liver fluke tetraspanin impairs parasite surface biogenesis and extracellular vesicle uptake by human host cells" (2025). GW Authored Works. Paper 8001.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/8001

Department

Microbiology, Immunology, and Tropical Medicine