Genomewide Analysis of Mode of Action of the S -Adenosylmethionine Analogue Sinefungin in Leishmania infantum

Abstract

The two main cellular metabolic one-carbon donors are reduced folates and S-adenosylmethionine, whose biosynthetic pathways have proven highly effective in chemotherapeutic interventions in various cell types. Sinefungin, a nucleoside analogue of S-adenosylmethionine, was shown to have potent activity against the protozoan parasite Leishmania. Here, we studied resistance to sinefungin using whole-genome approaches as a way to further our understanding of the role of S-adenosylmethionine in this parasite and to reveal novel potential drug targets. These approaches allowed the characterization of novel features related to S-adenosylmethionine function in Leishmania which could further help in the development of sinefungin-like compounds against this pathogenic parasite.

Experimental Strategies to Explore Drug Action and Resistance in Kinetoplastid Parasites

“…‘Omics’ approaches offer tools to identify drug targets in an untargeted manner in an either direct (affinity) or indirect approach, resulting in the generation of a (long) list of potential target candidates. Although this may seem to be a disadvantage, combining results of complementary techniques permits fine-tuning of acquired data [ 18 , 30 , 56 , 114 , 232 ]. Target validation by either chemical or genetic means can further confirm a potential role in the MoA [ 19 , 120 , 233 ].…”

“…Resistant strains are generally obtained upon in vitro selection on axenic parasites by stepwise increase of drug pressure. In some cases, mutagenesis prior to selection may generate more resistant mutants [ 26 , 56 , 57 , 58 , 59 , 60 ]. Selection on axenic stages has the advantage of being simple and relatively fast but lacks biological relevance in T. cruzi and Leishmania as these are intracellular pathogens in the vertebrate host.…”

“…The downside of experimental resistance selection is that a lot of spontaneous mutations occur because of the high continuous drug pressure or the prior mutagenesis, further complicating the identification of the actual gene(s) responsible for the loss in drug susceptibility [ 50 , 63 ]. The use of multiple parallel resistant strains could help in finding the needle in the haystack by identifying commonly mutated gene(s) [ 27 , 32 , 46 , 55 , 56 , 64 , 65 ].…”

Characterization of the NiRAN domain from RNA-dependent RNA polymerase provides insights into a potential therapeutic target against SARS-CoV-2

“…While Sorafernib and Sunitinib as approved for medical use in cases of renal, hepatocellular and gastro-intestinal cancers, the compound SU6656 is an experimental molecule used to study the role of Src kinases in cell cycle [56,57]. In addition to the kinase inhibitors, the compound 65482 (Sinefungin), is broad specificity microbial nucleotidyl transferase inhibitor, and is known to inhibit RNA replication in flaviviruses and herpes viruses [58,59]. The compound 122108 (Ribavirin 5’-triphosphate), inhibits the formation of g-capping of RNA in a wide range of viruses, such as Dengue virus, Hantaan virus and Hepatitis C virus [8,60,61].…”

Antioxidant Function and Metabolomics Study in Mice after Dietary Supplementation with Methionine

“…Methionine is an essential amino acid and mainly contributes to protein and S-adenosylmethionine (SAM) synthesis in the liver. SAM serves as a versatile methyl donor for nucleic acids and proteins, and the methylation of DNA and proteins is a major regulating mechanism for various physiological functions, including heat shock response [ 1 , 2 ], regulation of intestinal barrier integrity [ 3 ], development [ 4 ], and gene expression [ 5 ]. Meanwhile, methionine has been widely demonstrated to involve in oxidative stress via the transsulfuration pathway for cysteine generation [ 6 ].…”