Metabolomics-based studies in the field of Leishmania/leishmaniasis

Peer review under responsibility of Alexandria University Faculty of Medicine.

Available online 2 July 2018

Figures & data

Fig. 1 Different forms of Leishmania parasites during their infectious cycle. Procyclic promastigotes exist in the gut of sand-fly vector and divide quickly. Then this form develops into the metacyclic promastigotes forms. Transformation of procyclic to metacyclic is named metacyclogenesis process. In mammalian hosts metacyclic transforms in to intracellular amastigote forms in macrophage cells.

Fig. 2 The metabolomics-based study of Leishmania parasite process for NMR and LC-MS analysis. Sample preparation step of cells for NMR and MS-base analysis is different from each other which applying appropriate protocol could be obtain better results. The study of the metabolome of Leishmania-infected cells using metabolomics, can lead to the identification of metabolite targets for chemotherapeutic treatment and introducing of novel molecular markers that can be used for the diagnosis of different diseases.

Fig. 3 A summary of the most common analytical techniques (with advantages and disadvantages) used in metabolomics.

Table 1 Summary of ¹H NMR-based metabolomics studies conducted in the field of leishmania research.

Sample (Method)	Study Target	Metabolites/Metabolic Pathways	Ref.
L. major (^1H NMR)	Examination of the metabolites changes in the metacyclogenesis process (procyclic VS metacyclic)	Valine, leucine, and isoleucine biosynthesis: Citraconic acid, isopropylmalic acid, L-leucine Pyruvate metabolism: Acetic acid, isopropanol Glutathione metabolism: Ascorbic acid, ornithine Fatty acid biosynthesis: Caprylic acid, capric acid Sulfur metabolism: Acetic acid	33
L. major (^1H NMR)	Assessment of Artemisinin drug on metabolites profile (control VS case)	Galactose metabolism: α-D-glucose 6-phosphate, melibiose, raffinose, lactose staciose starch and sucrose metabolism: Amylase, α-D-glucose 6-phosphate sphingolipid metabolism: Sphingosine, Sphingaine valine, leucine, isoleucine biosynthesis: Threonine biotin metabolism : pimelate, 6-carboxyhexanol coA	36
L. major (^1H NMR)	Metabolic Characterization of L. major Infection in Activated and Nonactivated Macrophages	Discriminating Metabolites Associated with L. major Infection: Cell culture supernatants: Acetate, Alanine, Citrate, Fumarate, Glucose, Lactate, Ornithine, Pyruvate, Succinate Cell Extracts: Acetate, β-Alanine, Citrate, Creatine, Creatine Phosphate, Glycerophosphocholine, Glycine, Lactate, Phosphocholine, Taurine Discriminating Metabolites Associated with Macrophage Activation: Cell culture supernatants: Acetate, Alanine, Citrate, Glucose, Glycine, Lactate, Ornithine, Pyruvate, Succinate Cell Extracts: Acetate, β-Alanine, Citrate, Creatine, Creatine Phosphate, Glucose, Glycerophosphocholine, Glycine, Lactate, Phosphocholine, Taurine	37
L. donovani (^1H NMR)	Comparing of axenic amastigotes metabolites with those of promastigotes and intracellular amastigotes	Common metabolites in three stages: succinate, lactate, α-glycerophosphoryl choline (α-GPC), betaine, arginine, alanine, valine Promastigote and axenic amastigte metabolites: βhydroxybutyrate Axenic and intracellular amastigote metabolites: acetoacetate	38
Blood serum (^1H NMR) (atomic absorption)	Metabonomic Study on Samples of Cutaneous Leishmaniasis and its Correlation with the Selenium Level in the Blood Serum	Threonine, Lipid, Lactate, Alanine	39

¹H NMR; one dimensional proton nuclear magnetic resonance.

Table2 Summary of MS-based metabolomics studies conducted in the field of leishmania research.

Leishmania spp. (Method)	Study target	Metabolites	Ref.
L. Mexicana (^*LC-MS) (^**GC-MS)	Elucidation of metabolic adaptation in mutant model of Leishmania	Glycerol, Nicotinate, Phosphoethanol-amine, Glutamate 5-semialdehyde, Ovothiol A disulfide, Cytosine, Phosphocholine, Betaine, 2-Oxoglutaramate, Dihydroorotate, Deoxyribose, L-Glutamate, L-Aspartate, L-Alanine, Glutathione, Succinate	41
L. donovani L. mexicana L. major (LC-MS)	Comparative analysis of the metabolomes of promastigotes of L. donovani, L. major and L. mexicana	Differentiated confirmed metabolites between the cell extracts of three species: 64 metabolites Differentiated confirmed metabolites between the cell extracts of three species: 43 metabolites Number of confirmed and putative metabolites: 355 metabolites Large differences metabolites : tryptophan, aspartate, arginine and proline	42
L. infantum (LC-MS)	Elucidation of the metabolic changes involved in miltefosine action on JPCM5 promastigote cell line	Over 800 metabolites detected, 10% of which were significantly altered after 3.75 h(many of the changes related to an increase in alkane fragment and sugar release)	43
L. donovani (LC-MS)	Identification of metabolic differences on three different levels: (i) during in vitro promastigote growth; (ii) between strains with a different drug susceptibility; and (iii) when a resistant strain is maintained under drug pressure	Differential Significant decreased metabolites between BPK275_SbIII and BPK275: LOG: butyryl-, capryloyl-, valerylglycine, 2 fatty acyls, 2-oxoarginine and acetyllysine STAT: Xanthosine 5′-phosphate, trypanothione, low unsaturated GPL Differential metabolites change in resistant VS. sensitive strains growth stages: Up LOG R/LOG S: GPL building blocks Up STAT R/STAT S: Unsaturated GPLs, Amino acids, Purine nucleosides, Sugar phosphates Down LOG R/LOG S: Nucleobases and nucleosides Down STAT R/STAT S: Saturated GPLs	44
L. donovani (LC-MS)	Untargeted metabolite profiling and comparison their levels between L. donovani Promastigotes during in vitro culturing from logarithmic to stationary phase	Alchols, amines, amino acids, carbohydrates, fatty acyls, glycerolipids, glycerophospholipids, heterocyclic moleculs, ketones and aldehydes, nucleosides, organic acids, SL and sphingoid bases, sterol and prenol lipids, vitamins and cofactors	45
L. donovani (LC-MS)	Characterization of the global metabolic differences between antimonial-sensitive and antimonial-resistant L. donovani isolates	Glycerophospholipids, sphingolipids and bases, glycerolipids, sterol and prenol lipids, amino acids, polypeptides and conjugates, purines/ pyrimidines and conjugates, miscellaneous	46

L; Leishmania, LC-MS; liquid chromatography, GC-MS; gas chromatography, LOG; logarithmic growth stage, STAT; stationary growth stage, up; significant increase, down; significant decrease.

* Untargeted metabolomics platform.

** Targeted metabolomics platform.