Genotypic Status of the TbAT1/P2 Adenosine Transporter of Trypanosoma Brucei Gambiense Isolates from Northwestern Uganda Following Melarsoprol Withdrawal

Overview

Journal PLoS Negl Trop Dis

Specialties Infectious Diseases
Tropical Medicine

Date 2009 Sep 30

PMID 19787038

Citations 9

Authors

Anne J N Kazibwe

Barbara Nerima

Harry P de Koning

Pascal Maser

Michael P Barrett

Enock Matovu

Affiliations

Soon will be listed here.

Abstract

Background: The development of arsenical and diamidine resistance in Trypanosoma brucei is associated with loss of drug uptake by the P2 purine transporter as a result of alterations in the corresponding T. brucei adenosine transporter 1 gene (TbAT1). Previously, specific TbAT1 mutant type alleles linked to melarsoprol treatment failure were significantly more prevalent in T. b. gambiense from relapse patients at Omugo health centre in Arua district. Relapse rates of up to 30% prompted a shift from melarsoprol to eflornithine (alpha-difluoromethylornithine, DFMO) as first-line treatment at this centre. The aim of this study was to determine the status of TbAT1 in recent isolates collected from T. b. gambiense sleeping sickness patients from Arua and Moyo districts in Northwestern Uganda after this shift in first-line drug choice.

Methodology And Results: Blood and cerebrospinal fluids of consenting patients were collected for DNA preparation and subsequent amplification. All of the 105 isolates from Omugo that we successfully analysed by PCR-RFLP possessed the TbAT1 wild type allele. In addition, PCR/RFLP analysis was performed for 74 samples from Moyo, where melarsoprol is still the first line drug; 61 samples displayed the wild genotype while six were mutant and seven had a mixed pattern of both mutant and wild-type TbAT1. The melarsoprol treatment failure rate at Moyo over the same period was nine out of 101 stage II cases that were followed up at least once. Five of the relapse cases harboured mutant TbAT1, one had the wild type, while no amplification was achieved from the remaining three samples.

Conclusions/significance: The apparent disappearance of mutant alleles at Omugo may correlate with melarsoprol withdrawal as first-line treatment. Our results suggest that melarsoprol could successfully be reintroduced following a time lag subsequent to its replacement. A field-applicable test to predict melarsoprol treatment outcome and identify patients for whom the drug can still be beneficial is clearly required. This will facilitate cost-effective management of HAT in rural resource-poor settings, given that eflornithine has a much higher logistical requirement for its application.

Citing Articles

Genetic Diversity in the Diminazene Resistance-Associated P2 Adenosine Transporter-1 () Gene of .

Ashraf S, Yasein G, Ali Q, Afshan K, Betson M, Sargison N Animals (Basel). 2025; 15(5).

PMID: 40076038 PMC: 11898887. DOI: 10.3390/ani15050756.

Systematic Review and Meta-Analysis on Human African Trypanocide Resistance.

Kasozi K, MacLeod E, Welburn S Pathogens. 2022; 11(10).

PMID: 36297157 PMC: 9612373. DOI: 10.3390/pathogens11101100.

Current Treatments to Control African Trypanosomiasis and One Health Perspective.

Venturelli A, Tagliazucchi L, Lima C, Venuti F, Malpezzi G, Magoulas G Microorganisms. 2022; 10(7).

PMID: 35889018 PMC: 9321528. DOI: 10.3390/microorganisms10071298.

The Drugs of Sleeping Sickness: Their Mechanisms of Action and Resistance, and a Brief History.

de Koning H Trop Med Infect Dis. 2020; 5(1).

PMID: 31963784 PMC: 7157662. DOI: 10.3390/tropicalmed5010014.

The animal trypanosomiases and their chemotherapy: a review.

Giordani F, Morrison L, Rowan T, de Koning H, Barrett M Parasitology. 2016; 143(14):1862-1889.

PMID: 27719692 PMC: 5142301. DOI: 10.1017/S0031182016001268.

References

Maser P, Sutterlin C, Kralli A, Kaminsky R . A nucleoside transporter from Trypanosoma brucei involved in drug resistance. Science. 1999; 285(5425):242-4. DOI: 10.1126/science.285.5425.242. View

Becker S, Franco J, Simarro P, Stich A, Abel P, Steverding D . Real-time PCR for detection of Trypanosoma brucei in human blood samples. Diagn Microbiol Infect Dis. 2004; 50(3):193-9. DOI: 10.1016/j.diagmicrobio.2004.07.001. View

de Koning H, Anderson L, Stewart M, Burchmore R, Wallace L, Barrett M . The trypanocide diminazene aceturate is accumulated predominantly through the TbAT1 purine transporter: additional insights on diamidine resistance in african trypanosomes. Antimicrob Agents Chemother. 2004; 48(5):1515-9. PMC: 400564. DOI: 10.1128/AAC.48.5.1515-1519.2004. View

Geiser F, Luscher A, de Koning H, Seebeck T, Maser P . Molecular pharmacology of adenosine transport in Trypanosoma brucei: P1/P2 revisited. Mol Pharmacol. 2005; 68(3):589-95. DOI: 10.1124/mol.104.010298. View

De Koning H, Jarvis S . Uptake of pentamidine in Trypanosoma brucei brucei is mediated by the P2 adenosine transporter and at least one novel, unrelated transporter. Acta Trop. 2001; 80(3):245-50. DOI: 10.1016/s0001-706x(01)00177-2. View

Barrett M . The rise and fall of sleeping sickness. Lancet. 2006; 367(9520):1377-8. DOI: 10.1016/S0140-6736(06)68591-7. View

Cattand P, Miezan B, De Raadt P . Human African trypanosomiasis: use of double centrifugation of cerebrospinal fluid to detect trypanosomes. Bull World Health Organ. 1988; 66(1):83-6. PMC: 2491106. View

Barrett M . The fall and rise of sleeping sickness. Lancet. 1999; 353(9159):1113-4. DOI: 10.1016/S0140-6736(98)00416-4. View

Moore A, Richer M . Re-emergence of epidemic sleeping sickness in southern Sudan. Trop Med Int Health. 2001; 6(5):342-7. DOI: 10.1046/j.1365-3156.2001.00714.x. View

10.

Kaminsky R, Chuma F, Zweygarth E . Trypanosoma brucei brucei: expression of drug resistance in vitro. Exp Parasitol. 1989; 69(3):281-9. DOI: 10.1016/0014-4894(89)90074-x. View

11.

Checchi F, Piola P, Ayikoru H, Thomas F, Legros D, Priotto G . Nifurtimox plus Eflornithine for late-stage sleeping sickness in Uganda: a case series. PLoS Negl Trop Dis. 2007; 1(2):e64. PMC: 2100371. DOI: 10.1371/journal.pntd.0000064. View

12.

Stewart M, Krishna S, Burchmore R, Brun R, de Koning H, Boykin D . Detection of arsenical drug resistance in Trypanosoma brucei with a simple fluorescence test. Lancet. 2005; 366(9484):486-7. DOI: 10.1016/S0140-6736(05)66793-1. View

13.

Burri C, Keiser J . Pharmacokinetic investigations in patients from northern Angola refractory to melarsoprol treatment. Trop Med Int Health. 2001; 6(5):412-20. DOI: 10.1046/j.1365-3156.2001.00725.x. View

14.

Moser D, Cook G, Ochs D, Bailey C, McKane M, Donelson J . Detection of Trypanosoma congolense and Trypanosoma brucei subspecies by DNA amplification using the polymerase chain reaction. Parasitology. 1989; 99 Pt 1:57-66. DOI: 10.1017/s0031182000061023. View

15.

Morrison L, McCormack G, Sweeney L, Likeufack A, Truc P, Turner C . Use of multiple displacement amplification to increase the detection and genotyping of trypanosoma species samples immobilized on FTA filters. Am J Trop Med Hyg. 2007; 76(6):1132-7. PMC: 2067248. View

16.

Lanteri C, Stewart M, Brock J, Alibu V, Meshnick S, Tidwell R . Roles for the Trypanosoma brucei P2 transporter in DB75 uptake and resistance. Mol Pharmacol. 2006; 70(5):1585-92. DOI: 10.1124/mol.106.024653. View

17.

Maina N, Maina K, Maser P, Brun R . Genotypic and phenotypic characterization of Trypanosoma brucei gambiense isolates from Ibba, South Sudan, an area of high melarsoprol treatment failure rate. Acta Trop. 2007; 104(2-3):84-90. DOI: 10.1016/j.actatropica.2007.07.007. View

18.

Brun R, Schumacher R, Schmid C, Kunz C, Burri C . The phenomenon of treatment failures in Human African Trypanosomiasis. Trop Med Int Health. 2001; 6(11):906-14. DOI: 10.1046/j.1365-3156.2001.00775.x. View

19.

Woo P . The haematocrit centrifuge technique for the diagnosis of African trypanosomiasis. Acta Trop. 1970; 27(4):384-6. View

20.

Matovu E, Enyaru J, Legros D, Schmid C, Seebeck T, Kaminsky R . Melarsoprol refractory T. b. gambiense from Omugo, north-western Uganda. Trop Med Int Health. 2001; 6(5):407-11. DOI: 10.1046/j.1365-3156.2001.00712.x. View