Rôle(s) du métabolisme du glycérol dans la biologie des trypanosomes Africains

<div> Trypanosoma brucei, an extracellular parasite responsible for African trypanosomiasis, must adapt to distinct environments in its mammalian hosts and the tsetse fly vector. In the mammalian bloodstream, glucose serves as the primary carbon source, fueling the parasite's central carbon metabolism and ATP production, which supports its rapid growth. Once the parasites reach high cell densities, a quorumsensing mechanism induces a transition from proliferative slender forms to growtharrested stumpy forms (stumpy-QS). These stumpy forms help prevent host mortality by limiting parasitaemia and are primed for transmission to the tsetse fly. However, it has been demonstrated that glycerol can effectively replace glucose in feeding the parasite's central carbon metabolism, suggesting a significant role in vivo. This aligns with findings that trypanosomes predominantly reside in the extravascular spaces of tissues such as the skin and adipose tissue, where interstitial glycerol concentrations are 5 to 20 times higher than in plasma. Glycerol is released from adipocytes through both lipolysis and lipolysis-independent processes such as glycolysis, and it has been suggested that trypanosome-induced adipocyte lipolysis may even protect the host against trypanosome infection. Together, these data suggest that interactions between adipocytes and trypanosomes, potentially mediated by glycerol, play a critical role in the parasite's life cycle. This thesis explores the impact of glycerol on bloodstream form (BSF) Trypanosoma brucei. Our findings demonstrated that glycerol induces the differentiation of slender BSF into growth-arrested forms that resemble stumpy-QS, but with enhanced survival. Furthermore, under tissue-like conditions, characterized by glycerol levels between 0.2-0.5 mM and glucose at 4 mM, proliferative intermediate forms were generated, which were capable of differentiating into the insect vector stage (procyclics) and sustaining infections in tsetse flies. Additionally, glycerol extended the lifespan of quorum-sensing-induced stumpy forms, which normally have a limited lifespan of a few days. All these data led us to propose a revised model for transmission, in which quorum sensing-induced stumpy-QS forms protect the host from high parasitaemia, while glycerol from adipocytes induces intermediate-Glyc or long-lived stumpy forms that facilitate transmission to the fly. </div> <div>v Another key aspect of my thesis concerns the dissection of the signaling pathway involved in glycerol-induced differentiation. By exploiting the extended lifespan of stumpy-Glyc cells in culture, we selected mutants resistant to glycerol-induced differentiation through extended in vitro culturing in a glycerol-containing medium. Comparative genomic analyses between these mutants and cells grown in glucose, which are sensitive to glycerol-induced differentiation, identified candidate mutations associated with the resistance phenotype. Notably, these mutations were found to affect the protein kinase A regulatory subunit (PKAR), whose role in the signaling pathway was validated. Finally, we explored whether T. brucei can metabolize glycerol secreted by adipocytes even in the presence of excess glucose. To investigate this, we used an in vitro coculture system using a transwell assay, which allowed us to analyse the interactions between parental and mutant trypanosomes and adipocytes. We examined growth and exometabolome profiles using nuclear magnetic resonance (NMR)-based metabolite profiling, coupled with 13 C-labeling to trace specific metabolites. Our data showed that T. brucei efficiently utilized glycerol secreted by adipocytes to support its central carbon metabolism, even when glucose was abundant. Together, these data demonstrated that glycerol is a key player in the biology of Trypanosoma brucei. </div>