Quantitative proteomics of Trypanosoma cruzi during metacyclogenesis

Abstract

Trypanosoma cruzi is the etiologic agent of Chagas disease, which is estimated to affect over eight million people around the world. Trypanosoma cruzi has a complex life cycle, involving insect and mammalian hosts and four distinct developmental stages: epimastigotes, metacyclic trypomastigotes, amastigotes, and bloodstream trypomastigotes. Metacyclogenesis is the process by which T. cruzi epimastigotes differentiate into metacyclic trypomastigotes and acquire infectivity, and involves differential gene expression associated with acquisition of virulence. In T. cruzi, gene expression regulation is achieved mainly posttranscriptionally. Therefore, proteomics-based approaches are extremely useful for gaining a better understanding of the changes that occur in the stage-regulated gene expression program of the parasite at the molecular level. Here, we performed an in-depth quantitative MS-based proteomic study of T. cruzi metacyclogenesis and quantified almost 3000 proteins expressed during the process of differentiation. To the best of our knowledge, this work is the most comprehensive quantitative proteomics study of different cell populations of T. cruzi available so far. We identified relevant proteins and pathways involved in the parasite's differentiation and infectivity acquisition, opening new perspectives for further studies that could, ultimately, lead to the identification of new targets for chemotherapy.

Metabolomic profiling reveals a finely tuned, starvation-induced metabolic switch in Trypanosoma cruzi epimastigotes

“…4 and supplemental Table S2). By con- 5 The abbreviations used are: EPE, exponential phase epimastigote; SPE, stationary phase epimastigote; TCA, tricarboxylic acid; P5C, ⌬ 1 -pyrroline-5-carboxylate; P5CDH, P5C dehydrogenase; TSH, trypanothione; RTP, reverse trans-sulfuration pathway; MRM, multiple-reaction monitoring; PCA, principal component analysis; BCAA, branched-chain amino acid; ␥-GS, ␥-glutamate semi-aldehyde; CBS, cystathionine ␤-synthase; Pyr, pyruvate; ␣-KG, ␣-ketoglutarate; ESI, electrospray ionization. trast, ␣-ketoglutarate (␣-KG) was the only derivative from the amino acid oxidation that appeared to exhibit around six times higher abundance in the SPE.…”

“…In addition, a previous work comparing exponential and stationary phases of T. cruzi epimastigotes (EPE 5 and SPE, respectively) has assessed the physiological changes occurring at each stage and how some genes are regulated during the epimastigote growth to understand their possible participation in the metacyclogenesis (4 -6). However, little is known about metabolic changes that occur along with this physiological transition, and there are no reported studies supporting a broader view of the adaptive changes that epimastigotes undergo before they develop into metacyclic trypomastigote stage.…”

Comprehensive glycoprofiling of the epimastigote and trypomastigote stages of Trypanosoma cruzi

“…It is known that the adaptation to distinct environments (e.g. the insect's digestive tract and the interior of mammalian cells), with different temperatures, nutrient availability and immune response, requires major molecular and cellular changes of T. cruzi [4][5][6].…”

Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis

“…However, metacyclogenesis can be mimicked in vitro using well-defined media that stimulate the differentiation process by simulating the vector's gut physicochemical conditions (10). This process results in important changes that have been studied intensively through proteomic approaches (11,12).…”