Cloning and Characterization of Trypanosoma congolense and T. vivax Nucleoside Transporters Reveal the Potential of P1-Type Carriers for the Discovery of Broad-Spectrum Nucleoside-Based Therapeutics against Animal African Trypanosomiasis

Abstract

African Animal Trypanosomiasis (AAT), caused predominantly by <i>Trypanosoma brucei brucei</i>, <i>T. vivax</i> and <i>T. congolense</i>, is a fatal livestock disease throughout Sub-Saharan Africa. Treatment options are very limited and threatened by resistance. Tubercidin (7-deazaadenosine) analogs have shown activity against individual parasites but viable chemotherapy must be active against all three species. Divergence in sensitivity to nucleoside antimetabolites could be caused by differences in nucleoside transporters. Having previously characterized the <i>T. brucei</i> nucleoside carriers, we here report the functional expression and characterization of the main adenosine transporters of <i>T. vivax</i> (TvxNT3) and <i>T. congolense</i> (TcoAT1/NT10), in a <i>Leishmania mexicana</i> cell line ('SUPKO') lacking adenosine uptake. Both carriers were similar to the <i>T. brucei</i> P1-type transporters and bind adenosine mostly through interactions with N3, N7 and 3'-OH. Expression of TvxNT3 and TcoAT1 sensitized SUPKO cells to various 7-substituted tubercidins and other nucleoside analogs although tubercidin itself is a poor substrate for P1-type transporters. Individual nucleoside EC₅₀s were similar for <i>T. b. brucei</i>, <i>T. congolense</i>, <i>T. evansi</i> and <i>T. equiperdum</i> but correlated less well with <i>T. vivax</i>. However, multiple nucleosides including 7-halogentubercidines displayed pEC50>7 for all species and, based on transporter and anti-parasite SAR analyses, we conclude that nucleoside chemotherapy for AAT is viable.