Metabolomic Reprogramming of C57BL/6-Macrophages during Early Infection with L. amazonensis

Abstract

Leishmania survival inside macrophages depends on factors that lead to the immune response evasion during the infection. In this context, the metabolic scenario of the host cell–parasite relationship can be crucial to understanding how this parasite can survive inside host cells due to the host’s metabolic pathways reprogramming. In this work, we aimed to analyze metabolic networks of bone marrow-derived macrophages from C57BL/6 mice infected with Leishmania amazonensis wild type (La-WT) or arginase knocked out (La-arg−), using the untargeted Capillary Electrophoresis-Mass Spectrometry (CE-MS) approach to assess metabolomic profile. Macrophages showed specific changes in metabolite abundance upon Leishmania infection, as well as in the absence of parasite-arginase. The absence of L. amazonensis-arginase promoted the regulation of both host and parasite urea cycle, glycine and serine metabolism, ammonia recycling, metabolism of arginine, proline, aspartate, glutamate, spermidine, spermine, methylhistidine, and glutathione metabolism. The increased L-arginine, L-citrulline, L-glutamine, oxidized glutathione, S-adenosylmethionine, N-acetylspermidine, trypanothione disulfide, and trypanothione levels were observed in La-WT-infected C57BL/6-macrophage compared to uninfected. The absence of parasite arginase increased L-arginine, argininic acid, and citrulline levels and reduced ornithine, putrescine, S-adenosylmethionine, glutamic acid, proline, N-glutamyl-alanine, glutamyl-arginine, trypanothione disulfide, and trypanothione when compared to La-WT infected macrophage. Moreover, the absence of parasite arginase leads to an increase in NO production levels and a higher infectivity rate at 4 h of infection. The data presented here show a host-dependent regulation of metabolomic profiles of C57BL/6 macrophages compared to the previously observed BALB/c macrophages infected with L. amazonensis, an important fact due to the dual and contrasting macrophage phenotypes of those mice. In addition, the Leishmania-arginase showed interference with the urea cycle, glycine, and glutathione metabolism during host–pathogen interactions.

miR-294 and miR-410 Negatively Regulate Tnfa, Arginine Transporter Cat1/2, and Nos2 mRNAs in Murine Macrophages Infected with Leishmania amazonensis

“…The IL-1β and TNF levels also induced NO production [49]. Our group showed increased levels of arginine, citrulline, and polyamines inside the macrophages infected with L. amazonensis [50]. These data increased the modulation expectation of genes that can compete for arginine to produce polyamines, such as Cats, Arg1, Arg2, and Odc1.…”

Polyamines shift expression of macrophage L-arginine metabolism related-genes during Leishmania amazonensis infection

“…Metabolomic data reported by our group reveal increased levels of L-arginine, ornithine, putrescine, spermine and glutamine in BALB/c and C57BL/6 macrophages after 4h of infection with L. amazonensis (35,45). Infection of BALB/c macrophages with L. amazonensis knockout for arginase leads to the accumulation of L-arginine during infection, while proline, ornithine, and putrescine were diminished relative to infections with wild type parasite (35,45). These results reinforce that parasite arginase consumes L-arginine from the host, as described for amastigote forms consuming nutrients from phagolysosome, affecting the supply for ARG1 and NOS2 (46).…”

Polyamine Metabolism in Leishmania Parasites: A Promising Therapeutic Target

“…However, in L. donovani , only one of these gene copies, LdAAP3.2 , is regulated in response to changes in host arginine levels and notably, when deleted from the L. donovani genome, it impeded the infection of host macrophages and BALB/C mice [ 91 , 200 ]. This suggests that targeting of this transporter may be of therapeutic value, particularly considering that higher levels of host arginine have been linked to increased nitric oxide production and reduced parasite survival [ 80 , 102 , 206 ].…”