Differential expression of polyamine biosynthetic pathways in skin lesions and in plasma reveals distinct profiles in diffuse cutaneous leishmaniasis

Hayna Malta-Santos,Jackson M Costa,Amanda Macedo,Ricardo Khouri,Eny Iochevet Segal Floh,Kiyoshi F Fukutani,Bruno B Andrade,Johan Van Weyenbergh,Sandra Marcia Muxel,Jaqueline França-Costa,Aldina Barral,Artur T L Queiroz,Lucile M Floeter-Winter,Viviane Boaventura,Valéria M Borges

doi:10.1038/s41598-020-67432-5

Abstract

Tegumentary leishmaniasis (TL) is a parasitic disease that can result in wide spectrum clinical manifestations. It is necessary to understand host and parasite determinants of clinical outcomes to identify novel therapeutic targets. Previous studies have indicated that the polyamine biosynthetic pathway is critical for Leishmania growth and survival. Despite its importance, expression of the such pathway has not been previously investigated in TL patients. We performed an exploratory analysis employing Systems Biology tools to compare circulating polyamines and amino acid concentration as well as polyamine pathway gene expression in cutaneous lesions patients presenting with distinct TL disease presentations. Diffuse cutaneous leishmaniasis (DCL) was associated with higher concentrations of amino acids, polyamines and its substrate transporters than mucosal cutaneous leishmaniasis or localized cutaneous leishmaniasis. In addition, the RNA expression of polyamine-related genes of patients lesions from two separate cohorts demonstrated that differential activation of this pathway is associated with parasite loads and able to discriminate the clinical spectrum of TL. Taken together, our findings highlight a new aspect of DCL immunopathogenesis indicating that the polyamine pathway may be explored as a novel therapeutic target to control disease burden.

Highlights

Tegumentary leishmaniasis (TL) is a parasitic disease that can result in wide spectrum clinical manifestations
We have previously shown high concentrations of arginase-1 (ARG1), ornithine decarboxylase (ODC), prostaglandin E2 (PGE2) and transforming growth factor β (TGF-β) in Diffuse cutaneous leishmaniasis (DCL) patients[4], which could contribute to an ineffective immune response unable to hamper parasite replication
We observed that the relative systemic concentrations of arginase-1, cadaverine and spermidine, but not of putrescine, were significantly higher in DCL, compared with either localized cutaneous leishmaniasis (LCL), mucocutaneous leishmaniasis (MCL) patients or health controls (Fig. 1B, C and Table S1)

Summary

Results

DCL patients exhibit high plasma levels of polyamines and amino acids. Initially we tested if there is a distinct systemic profile of plasma concentrations of arginase-1 (protein), amino acids and free polyamines in patients with TL. To validate our findings on the polyamine pathway in TL, we re-analyzed the gene expression data from an independent patient cohort, which was recently p ublished[16] This approach revealed that skin lesions from TL patients generally exhibit a distinct gene expression profile compared to normal skin from uninfected healthy endemic controls (Fig. 3A). Hierarchical cluster analysis of the Leishmania transcripts quantified by the RNAseq indicated that patients with DCL were the ones who displayed the highest levels of parasite transcripts (Fig. 4A) Our analyses demonstrated that while expression values of several genes exhibited similar association profiles with parasite loads between DCL and LCL patient groups, such as ARG1, other targets displayed a divergent pattern, such as NOS1 and NOS3 (Fig. 4B). GCH1 SLC3A2 ASS1 OAT SLC7A1 SLC7A5 SRM SLC7A7 PYCR1 ASL AGMAT SAT1 NOS1 NOS3 ARG2 PTS AMD1 SLC7A2 SMS PAOX ARG1 ALDH5A1 fold-difference (log2)

Discussion

Parasite Load

Methods