Different Mutations in a P-type ATPase Transporter in Leishmania Parasites are Associated with Cross-resistance to Two Leading Drugs by Distinct Mechanisms

Abstract

Leishmania infantum is an etiological agent of the life-threatening visceral form of leishmaniasis. Liposomal amphotericin B (AmB) followed by a short administration of miltefosine (MF) is a drug combination effective for treating visceral leishmaniasis in endemic regions of India. Resistance to MF can be due to point mutations in the miltefosine transporter (MT). Here we show that mutations in MT are also observed in Leishmania AmB-resistant mutants. The MF-induced MT mutations, but not the AmB induced mutations in MT, alter the translocation/uptake of MF. Moreover, mutations in the MT selected by AmB or MF have a major impact on lipid species that is linked to cross-resistance between both drugs. These alterations include changes of specific phospholipids, some of which are enriched with cyclopropanated fatty acids, as well as an increase in inositolphosphoceramide species. Collectively these results provide evidence of the risk of cross-resistance emergence derived from current AmB-MF sequential or co-treatments for visceral leishmaniasis.

Chemogenomic profiling of anti-leishmanial efficacy and resistance in the related kinetoplastid parasite Trypanosoma brucei

“…Consistent with this, T. brucei RNAi library selection in miltefosine led to enrichment for RNAi fragments mapping to the syntenic sequence in T. brucei ( Tb927.11.3350 ). RNAi library selection in amphotericin-B also enriched for RNAi fragments mapping to this gene, consistent with recent findings in Leishmania (46), as well as two other flippases and a putative β-subunit (Tb927.11.13200). Interestingly, the β-subunit targeted was not the syntenic orthologue of Ros3, previously shown to interact with the MT (18).…”

“…This is consistent with disruption of membranes by amphotericin-B. Miltefosine uptake in Leishmania is dependent on a flippase (17, 18), which also contributes to the anti-leishmanial action of amphotericin-B (46). RNAi fragments targeting the syntenic locus in T. brucei , Tb927.11.3350 , were enriched following selection in amphotericin-B and miltefosine (Fig.…”

Leishmania dual‐specificity tyrosine‐regulated kinase 1 (DYRK1) is required for sustaining Leishmania stationary phase phenotype

“…Tandem mass spectra (MS/MS) were obtained for precursors of phosphatidylinositol (PI)/inositol–phosphoceramide (IPC) species in negative ion mode, with parent‐ion scanning of m / z 241 (collision energy of 70V); and phosphatidylcholine (PC)/sphingomyelin (SM) species in positive mode, with parent ion scanning of m / z 184 (collision energy of 50V). Assignment of phospholipid species is based upon a combination of survey, daughter and precursor scans, as well as previous assignments (Fernandez‐Prada et al, ; Richmond et al, ) and use of the LIPID MAPS: Nature Lipidomics Gateway (://www.lipidmaps.org).…”

“…Mass spectra were acquired from 50–550 amu. Identification was carried out by comparison of the retention times and fragmentation patterns with sterols in the NIST/EPA/NIH Mass Spectral Library and those reported in previous publications (Cauchetier et al, ; Fernandez‐Prada et al, ; Yao & Wilson, ). To examine changes in composition, the area under each sterol peak was used for relative quantification.…”

Comparative Transcriptomic Analysis of Antimony Resistant and Susceptible Leishmania Infantum Lines

“…ATPases are membrane proteins that perform active ion transport across biological membranes, which are found in bacteria and all eukaryotic cells, including Leishmania [89]. Fernandez-Prada et al (2016) demonstrated that different point mutations in a P-type ATPase transporter in L. infantum are implicated with cross-resistance to miltefosine and amphotericin B [90].…”