Knockout of liver fluke granulin, Ov-grn-1, impedes malignant transformation during chronic infection with Opisthorchis viverrini

Sujittra Chaiyadet, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Sirikachorn Tangkawattana, Faculty of Veterinary Medicine, Khon Kaen University, Khon Kaen, Thailand, and WHO Collaborating Center for Research and Control of Opisthorchiasis, Tropical Disease Research Center, Khon Kaen University, Khon Kaen, Thailand.
Michael J. Smout, Centre for Molecular Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia.
Wannaporn Ittiprasert, Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine & Health Sciences, George Washington University, Washington, District of Columbia, United States of America.
Victoria H. Mann, Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine & Health Sciences, George Washington University, Washington, District of Columbia, United States of America.
Raksawan Deenonpoe, Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Patpicha Arunsan, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Alex Loukas, Centre for Molecular Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, Queensland, Australia.
Paul J. Brindley, Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, School of Medicine & Health Sciences, George Washington University, Washington, District of Columbia, United States of America.
Thewarach Laha, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.

Document Type

Journal Article

Publication Date

9-22-2022

Journal

PLoS pathogens

Volume

18

Issue

9

DOI

10.1371/journal.ppat.1010839

Abstract

Infection with the food-borne liver fluke Opisthorchis viverrini is the principal risk factor for cholangiocarcinoma (CCA) in the Mekong Basin countries of Thailand, Lao PDR, Vietnam, Myanmar and Cambodia. Using a novel model of CCA, involving infection with gene-edited liver flukes in the hamster during concurrent exposure to dietary nitrosamine, we explored the role of the fluke granulin-like growth factor Ov-GRN-1 in malignancy. We derived RNA-guided gene knockout flukes (ΔOv-grn-1) using CRISPR/Cas9/gRNA materials delivered by electroporation. Genome sequencing confirmed programmed Cas9-catalyzed mutations of the targeted genes, which was accompanied by rapid depletion of transcripts and the proteins they encode. Gene-edited parasites colonized the biliary tract of hamsters and developed into adult flukes. However, less hepatobiliary tract disease manifested during chronic infection with ΔOv-grn-1 worms in comparison to hamsters infected with control gene-edited and mock-edited parasites. Specifically, immuno- and colorimetric-histochemical analysis of livers revealed markedly less periductal fibrosis surrounding the flukes and less fibrosis globally within the hepatobiliary tract during infection with ΔOv-grn-1 genotype worms, minimal biliary epithelial cell proliferation, and significantly fewer mutations of TP53 in biliary epithelial cells. Moreover, fewer hamsters developed high-grade CCA compared to controls. The clinically relevant, pathophysiological phenotype of the hepatobiliary tract confirmed a role for this secreted growth factor in malignancy and morbidity during opisthorchiasis.

Department

Microbiology, Immunology, and Tropical Medicine

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