Silencing of Tetraspanin Gene Expression Results in Reduced Secretion of Extracellular Vesicles

Authors

Sujittra Chaiyadet, Tropical Medicine Graduate Program, Academic Affairs, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Javier Sotillo, Parasitology Reference and Research Laboratory, National Centre for Microbiology, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain.
Watchara Krueajampa, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Sophita Thongsen, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Michael Smout, Centre for Molecular Therapeutics, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns, QLD, Australia.
Paul J. Brindley, Department of Microbiology, Immunology and Tropical Medicine, and Research Center for Neglected Diseases of Poverty, George Washington University, Washington, DC, United States.
Thewarach Laha, 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, QLD, Australia.

Document Type

Journal Article

Publication Date

1-1-2022

Journal

Frontiers in cellular and infection microbiology

Volume

12

DOI

10.3389/fcimb.2022.827521

Keywords

Opisthorchis viverrini; endocytosis; extracellular vesicle; liver fluke; tetraspanin

Abstract

Inter-phylum transfer of molecular information is exquisitely exemplified in the uptake of parasite extracellular vesicles (EVs) by their target mammalian host tissues. The oriental liver fluke, is the major cause of bile duct cancer in people in Southeast Asia. A major mechanism by which promotes cancer is through the secretion of excretory/secretory products which contain extracellular vesicles (EVs). EVs contain microRNAs that are predicted to impact various mammalian cell proliferation pathways, and are internalized by cholangiocytes that line the bile ducts. Upon uptake, EVs drive relentless proliferation of cholangiocytes and promote a tumorigenic environment, but the underlying mechanisms of this process are unknown. Moreover, purification and characterization methods for helminth EVs in general are ill defined. We therefore compared different purification methods for EVs and characterized the sub-vesicular compartment proteomes. Two CD63-like tetraspanins (-TSP-2 and TSP-3) are abundant on the surface of EVs, and could serve as biomarkers for these parasite vesicles. Anti-TSP-2 and -TSP-3 IgG, as well as different endocytosis pathway inhibitors significantly reduced EV uptake and subsequent proliferation of cholangiocytes . Silencing of and 3 gene expression in adult flukes using RNA interference resulted in substantial reductions in EV secretion, and those vesicles that were secreted were deficient in their respective TSP proteins. Our findings shed light on the importance of tetraspanins in fluke EV biogenesis and/or stability, and provide a conceivable mechanism for the efficacy of anti-tetraspanin subunit vaccines against a range of parasitic helminth infections.

APA Citation

Chaiyadet, Sujittra; Sotillo, Javier; Krueajampa, Watchara; Thongsen, Sophita; Smout, Michael; Brindley, Paul J.; Laha, Thewarach; and Loukas, Alex, "Silencing of Tetraspanin Gene Expression Results in Reduced Secretion of Extracellular Vesicles" (2022). GW Authored Works. Paper 302.
https://hsrc.himmelfarb.gwu.edu/gwhpubs/302

Department

Microbiology, Immunology, and Tropical Medicine