Divergent metabolism between Trypanosoma congolense and Trypanosoma brucei results in differential drug sensitivity

Steketee, P. C.; Dickie, E. A.; Iremonger, J.; Crouch, K.; Paxton, E.; Jayaraman, S.; Alfituri, O. A.; Awuah-Mensah, G.; Ritchie, R.; Schnaufer, A.; De Koning, H.; Gadelha, C.; Wickstead, B.; Barrett, M. P.; Morrison, L. J.

doi:10.1101/2021.01.05.425368

bioRxiv · 10.1101/2021.01.05.425368

Divergent metabolism between Trypanosoma congolense and Trypanosoma brucei results in differential drug sensitivity

Steketee, P. C.·Dickie, E. A.·Iremonger, J.·Crouch, K.·Paxton, E.·Jayaraman, S.·Alfituri, O. A.·Awuah-Mensah, G.·Ritchie, R.·Schnaufer, A.·De Koning, H.·Gadelha, C.·Wickstead, B.·Barrett, M. P.·Morrison, L. J.

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Abstract

Animal African Trypanosomiasis (AAT) is a debilitating livestock disease prevalent across sub-Saharan Africa, a main cause of which is the protozoan parasite Trypanosoma congolense. In comparison to the well-studied T. brucei, there is a major paucity of knowledge regarding the biology of T. congolense. Here, we use a combination of omics technologies and novel genetic tools to characterise core metabolism in T. congolense mammalian-infective bloodstream-form parasites, and test whether metabolic differences compared to T. brucei impact upon drug sensitivity. Like T. brucei, glycolysis plays a major part in T. congolense energy metabolism. However, the rate of glucose uptake is significantly reduced in T. congolense, with cells remaining viable when cultured in concentrations as low as 2 mM. Instead of pyruvate, the primary glycolytic endpoints are succinate, malate and acetate. Comparative transcriptomics analysis showed higher levels of activity associated with the mitochondrial pyruvate dehydrogenase complex, acetate generation and the succinate shunt in T. congolense. However, based on omics analysis and chemical inhibition, there does not appear to be significant levels of oxidative phosphorylation. Stable-isotope labelling of glucose enabled the comparison of carbon usage between T. brucei and T. congolense, highlighting differences in nucleotide and fatty acid metabolism. To validate the metabolic similarities and differences, both species were treated with pharmacological inhibitors, confirming a lack of essential electron transport chain activity in T. congolense, but increased sensitivity to inhibition of mitochondrial pyruvate import. Strikingly, T. congolense exhibited significant resistance to inhibitors of fatty acid synthesis, including a 780-fold greater EC50 against the lipase and fatty acid synthase inhibitor Orlistat, compared to T. brucei. These data highlight that bloodstream form T. congolense diverges from T. brucei in key areas of metabolism, with several features that are intermediate between bloodstream- and insect-stage T. brucei. These results have implications for drug development, mechanisms of drug resistance and host-pathogen interactions.

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Steketee, P. C., Dickie, E. A., Iremonger, J., Crouch, K., Paxton, E., Jayaraman, S., Alfituri, O. A., Awuah-Mensah, G., Ritchie, R., Schnaufer, A., De Koning, H., Gadelha, C., Wickstead, B., Barrett, M. P., Morrison, L. J.. 2021-01-05. Divergent metabolism between Trypanosoma congolense and Trypanosoma brucei results in differential drug sensitivity. https://doi.org/10.1101/2021.01.05.425368

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