Programmed knockout mutation of liver fluke granulin attenuates virulence of infection-induced hepatobiliary morbidity

Arunsan, Patpicha, Ittiprasert, Wannaporn, Smout, Michael J., Cochran, Christina J., Mann, Victoria H., Chaiyadet, Sujittra, Karinshak, Shannon E., Sripa, Banchob, Young, Neil David, Sotillo, Javier, Loukas, Alex, Brindley, Paul J., and Laha, Thewarach (2019) Programmed knockout mutation of liver fluke granulin attenuates virulence of infection-induced hepatobiliary morbidity. eLife, 8. e41463.

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Abstract

Infection with the food-borne liver fluke Opisthorchis viverrini is the principal risk factor (IARC Working Group on the Evaluation of Carcinogenic Risks to Humans, 2012) for cholangiocarcinoma (CCA) in the Lower Mekong River Basin countries including Thailand, Lao PDR, Vietnam and Cambodia. We exploited this link to explore the role of the secreted growth factor termed liver fluke granulin (Ov-GRN-1) in pre-malignant lesions by undertaking programmed CRISPR/Cas9 knockout of the Ov-GRN-1 gene from the liver fluke genome. Deep sequencing of amplicon libraries from genomic DNA of gene-edited parasites revealed Cas9-catalyzed mutations within Ov-GRN-1. Gene editing resulted in rapid depletion of Ov-GRN-1 transcripts and the encoded Ov-GRN-1 protein. Gene-edited parasites colonized the biliary tract of hamsters and developed into adult flukes, but the infection resulted in reduced pathology as evidenced by attenuated biliary hyperplasia and fibrosis. Not only does this report pioneer programmed gene-editing in parasitic flatworms, but also the striking, clinically-relevant pathophysiological phenotype confirms the role for Ov-GRN-1 in virulence morbidity during opisthorchiasis.

Item ID: 57090
Item Type: Article (Research - C1)
ISSN: 2050-084X
Keywords: Opisthorchis; Clonorchis; Liver fluke; cholangiocarcinoma; Ov-GRN-1
Copyright Information: CC Copyright Arunsan et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.
Funders: Thailand Research Fund (TRF), National Health and Medical Research Council of Australia (NHMRC)
Projects and Grants: TRF PHD/0111/2557, NHMRC APP1109829
Date Deposited: 13 Feb 2019 07:37
FoR Codes: 11 MEDICAL AND HEALTH SCIENCES > 1108 Medical Microbiology > 110899 Medical Microbiology not elsewhere classified @ 50%
11 MEDICAL AND HEALTH SCIENCES > 1101 Medical Biochemistry and Metabolomics > 110199 Medical Biochemistry and Metabolomics not elsewhere classified @ 50%
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