In-Depth Quantitative Proteomics Characterization of In Vitro Selected Miltefosine Resistance in Leishmania infantum

Abstract

Visceral leishmaniasis (VL) is a neglected disease caused by Leishmania parasites. Although significant morbidity and mortality in tropical and subtropical regions of the world are associated with VL, the low investment for developing new treatment measures is chronic. Moreover, resistance and treatment failure are increasing for the main medications, but the emergence of resistance phenotypes is poorly understood at the protein level. Here, we analyzed the development of resistance to miltefosine upon experimental selection in a L. infantum strain. Time to miltefosine resistance emergence was ~six months and label-free quantitative mass-spectrometry-based proteomics analyses revealed that this process involves a remodeling of components of the membrane and mitochondrion, with significant increase in oxidative phosphorylation complexes, particularly on complex IV and ATP synthase, accompanied by increased energy metabolism mainly dependent on β-oxidation of fatty acids. Proteins canonically involved in ROS detoxification did not contribute to the resistant process whereas sterol biosynthesis enzymes could have a role in this development. Furthermore, changes in the abundance of proteins known to be involved in miltefosine resistance such as ABC transporters and phospholipid transport ATPase were detected. Together, our data show a more complete picture of the elements that make up the miltefosine resistance phenotype in L. infantum.

ABC transporter inhibition by beauvericin partially overcomes drug resistance in Leishmania tropica

“…The precise factors contributing to ML treatment failure remain a subject of ongoing investigation. Emerging evidence underscores a multifaceted interplay of factors, placing substantial emphasis not only on reduced drug accumulation ( 16 , 63 , 64 ) but also on the bolstered drug efflux mechanisms ( 15 , 64 , 75 , 76 ). The investigation into fluorescence decay in ML-treated parasites demonstrated notable variations between LS-LT2 and MLR-LT2 ( Fig.…”

“…To address the growing problem of resistance, higher doses of ML were recommended to overcome it ( 14 ), which, unfortunately, led to a doubling of the relapse rate. Resistance to ML in Leishmania can be associated with complex changes in drug transport mechanisms, including both decreased drug influx and increased drug efflux ( 15 ). The miltefosine transporter (MLT), operating as a flippase enzyme, in conjunction with its associated protein, Ros3, is demonstrated to play a pivotal role in the emergence of ML resistance by exerting control over the translocation of ML across the parasitic membrane ( 16 ).…”

Improvement of Genetic Variation of cv Grand Naeni Bananas Through Gamma-Ray Irradiation and In Vitro Selection Using Fusaric Acid

“…To accelerate the banana plant breeding program it is necessary to be supported by technological breakthroughs, one of which is mutagenesis followed by in vitro selection. In vitro selection techniques have been shown to be effective in selecting somaclone populations in selective media so as to increase the probability of producing plants with the desired traits (Damayanti et al, 2002;Roostika et al, 2013;Rao & Sandhya, 2016;Akaeze et al, 2017;Makkewicz et al, 2017;Rajoriya et al, 2018;Sivakumar & Visalakshi, 2021;Elyazid et al, 2021;Ani et al, 2022;Saboia-Vahia et al, 2022) on explants in in vitro culture using an elicitor. One of the elicitors commonly used to obtain resistance to Fusarium wilt is filtrate from fusarium culture or using pure fusarium poison, namely fusaric acid.…”

“…Increasing genetic diversity in bananas can be done through mutations of gamma ray irradiation (Abdulhafiz et al, 2018;Due et al, 2019;Ramesh et al, 2019;Ali et al, 2020;Udaya et al, 2021;Penna et al, 2021;Hasim et al, 2021). Increasing genetic diversity through mutation induction can provide populations of somaclone variants that can help develop plant varieties that are resistant to biotic and abiotic factors (Roostika et al, 2013;Nikam et al, 2014;Rao et al, 2016;Akaeze et al, 2017;Hegde et al, 2019;Upadhyay et al, 2020;Mishra et al, 2021;Saboia-Vahia et al, 2022).…”

Análisis epidemiológico, genómico y proteómico de Leishmania infantum en Colombia

“…Saboia-Vahia y colaboradores en 2022 describieron los cambios cualitativos y cuantitativos presentados por una línea de L. infantum resistente a MIL; en estos parasito además de evidenciar cambios en la cinética de crecimiento de los promastigotes, se observó un aumento significativo en los complejos de fosforilación oxidativa, principalmente, complejo IV, junto con una mayor actividad en la β-oxidación de ácidos grasos y los transportadores ABC; con una reducida concentración de proteínas asociadas a desintoxicación de especies reactivas de oxígeno (ROS) [216] . Recientemente investigadores como Gutiérrez-Guarnizo y colaboradores han realizado estudios de proteómica enfocados a comprender el traductoma en Leishmania, generando importantes hipótesis como la adaptación preventiva y la optimización del metabolismo energético de los parásitos con fenotipos de resistencia [217,218] .…”