title: Induced resistance to methionyl-tRNA synthetase inhibitors in Trypanosoma brucei is due to overexpression of the target.
abstract: New classes of antiparasitic drugs active against Trypanosoma brucei are needed to combat human African trypanosomiasis. Inhibitors of methionyl-tRNA synthetase (MetRS) have excellent potential to be developed for this purpose (S. Shibata, J. R. Gillespie, A. M. Kelley, A. J. Napuli, Z. Zhang, K. V. Kovzun, R. M. Pefley, J. Lam, F. H. Zucker, W. C. Van Voorhis, E. A. Merritt, W. G. Hol, C. L. Verlinde, E. Fan, and F. S. Buckner, Antimicrob. Agents Chemother. 55:1982-1989, 2011). In order to assess the potential for resistance to develop against this new class of inhibitors, T. brucei cultures were grown in the presence of MetRS inhibitors or comparison drugs. Resistance up to ∼50 times the baseline 50% inhibitory concentration (IC50) was induced against a MetRS inhibitor after ∼120 days. A similar level of resistance to the clinical drug eflornithine was induced after ∼50 days and for pentamidine after ∼80 days. Thus, resistance was induced more slowly against MetRS inhibitors than against clinically used drugs. The parasites resistant to the MetRS inhibitor were shown to overexpress MetRS mRNA by a factor of 35 over the parental strain. Southern analysis indicated that the MetRS gene was amplified in the genome by nearly 8-fold. When injected into mice, the MetRS inhibitor-resistant parasites caused a reduced level of infection, indicating that the changes associated with resistance attenuated their virulence. This finding and the fact that resistance to MetRS inhibitors developed relatively slowly are encouraging for further development of this class of compounds. Published studies on other antitrypanosomal drugs have primarily shown that alterations in membrane transporters were the mechanisms responsible for resistance. This is the first published report of induced drug resistance in the African trypanosome due to overexpression of the target enzyme.
authors.affiliation: Department of Medicine, University of Washington, Seattle, Washington, USA.
authors.forename: Ranae M
authors.initials: RM
authors.lastname: Ranade
authors.forename: J Robert
authors.initials: JR
authors.lastname: Gillespie
authors.forename: Sayaka
authors.initials: S
authors.lastname: Shibata
authors.forename: Christophe L M J
authors.initials: CL
authors.lastname: Verlinde
authors.forename: Erkang
authors.initials: E
authors.lastname: Fan
authors.forename: Wim G J
authors.initials: WG
authors.lastname: Hol
authors.forename: Frederick S
authors.initials: FS
authors.lastname: Buckner
mesh_terms: D000634:Aminoquinolines / Q000494:pharmacology; D000818:Animals; D001483:Base Sequence; D004351:Drug Resistance / Q000235:genetics; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D008718:Methionine-tRNA Ligase / Q000037:antagonists & inhibitors* / Q000235:genetics*; D051379:Mice; D021261:Parasitic Sensitivity Tests; D012333:RNA, Messenger / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014346:Trypanosoma brucei brucei / Q000187:drug effects* / Q000201:enzymology* / Q000378:metabolism; D014353:Trypanosomiasis, African / Q000188:drug therapy / Q000469:parasitology
chemical_list: D000634:Aminoquinolines; D012333:RNA, Messenger; D014344:Trypanocidal Agents; C025152:aminoquinolone; D008718:Methionine-tRNA Ligase