Evaluation of Giardia lamblia thioredoxin reductase as drug activating enzyme and as drug target.

Leitsch, David; Müller, Joachim; Müller, Norbert (2016). Evaluation of Giardia lamblia thioredoxin reductase as drug activating enzyme and as drug target. International journal for parasitology. Drugs and drug resistance, 6(3), pp. 148-153. Elsevier 10.1016/j.ijpddr.2016.07.003

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The antioxidative enzyme thioredoxin reductase (TrxR) has been suggested to be a drug target in several pathogens, including the protist parasite Giardia lamblia. TrxR is also believed to catalyse the reduction of nitro drugs, e.g. metronidazole and furazolidone, a reaction required to render these compounds toxic to G. lamblia and other microaerophiles/anaerobes. It was the objective of this study to assess the potential of TrxR as a drug target in G. lamblia and to find direct evidence for the role of this enzyme in the activation of metronidazole and other nitro drugs. TrxR was overexpressed approximately 10-fold in G. lamblia WB C6 cells by placing the trxR gene behind the arginine deiminase (ADI) promoter on a plasmid. Likewise, a mutant TrxR with a defective disulphide reductase catalytic site was strongly expressed in another G. lamblia WB C6 cell line. Susceptibilities to five antigiardial drugs, i.e. metronidazole, furazolidone, nitazoxanide, albendazole and auranofin were determined in both transfectant cell lines and compared to wildtype. Further, the impact of all five drugs on TrxR activity in vivo was measured. Overexpression of TrxR rendered G. lamblia WB C6 more susceptible to metronidazole and furazolidone but not to nitazoxanide, albendazole, and auranofin. Of all five drugs tested, only auranofin had an appreciably negative effect on TrxR activity in vivo, albeit to a much smaller extent than expected. Overexpression of TrxR and mutant TrxR had hardly any impact on growth of G. lamblia WB C6, although the enzyme also exerts a strong NADPH oxidase activity which is a source of oxidative stress. Our results constitute first direct evidence for the notion that TrxR is an activator of metronidazole and furazolidone but rather question that it is a relevant drug target of presently used antigiardial drugs.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Parasitology
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)

UniBE Contributor:

Leitsch, David; Müller, Joachim and Müller, Norbert

Subjects:

600 Technology > 630 Agriculture
500 Science > 570 Life sciences; biology

ISSN:

2211-3207

Publisher:

Elsevier

Language:

English

Submitter:

Norbert Müller

Date Deposited:

06 Jun 2017 15:12

Last Modified:

17 Dec 2018 08:52

Publisher DOI:

10.1016/j.ijpddr.2016.07.003

PubMed ID:

27485086

Uncontrolled Keywords:

Antigiardial drugs; Giardia lamblia; Thioredoxin reductase

BORIS DOI:

10.7892/boris.98096

URI:

https://boris.unibe.ch/id/eprint/98096

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