Cellular and molecular biological analyses of nifurtimox resistance in Trypanosoma cruzi.
We induced nifurtimox resistance in both epimastigotes and tissue culture-derived trypomastigotes of several Trypanosoma cruzi strains. The magnitude of nifurtimox resistance was strain-dependent. A variety of karyotype changes occurred in the nifurtimox-resistant (NR) strains. Chromosome-specific DNA probes identified karyotype changes common to the NR strains that were moderately resistant to nifurtimox but not the NR strains that were highly resistant to nifurtimox. A marked increase in nifurtimox resistance in one NR strain was accompanied by a 100% increase in nuclear DNA mass and a 50% increase in kinetoplast DNA mass. These data suggest that nifurtimox resistance can be accompanied by a wide spectrum of DNA changes. Both trypanothione reductase and heat-shock proteins may modulate the effects of exposure of T. cruzi to nifurtimox. However, we did not detect qualitative or quantitative differences in these genes or their transcripts between the NR strains and the sensitive strains from which they were derived. An understanding of the spectrum of diversity in nifurtimox resistance at the cellular and molecular levels demonstrated in this report is critical in the development of drug therapies against Chagas' disease.
Nozaki
T
T
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA.
Engel
J C
JC
Dvorak
J A
JA
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan / Q000235:genetics*; D004351:Drug Resistance / Q000235:genetics; D005434:Flow Cytometry; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007621:Karyotyping; D008969:Molecular Sequence Data; D009247:NADH, NADPH Oxidoreductases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D009547:Nifurtimox / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D013045:Species Specificity; D014349:Trypanosoma cruzi / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development
D016054:DNA, Protozoan; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D009247:NADH, NADPH Oxidoreductases; C049608:trypanothione reductase; D009547:Nifurtimox