Comparative transcriptome profiling of virulent and non-virulent Trypanosoma cruzi underlines the role of surface proteins during infection.
Comparative transcriptome profiling of virulent and non-virulent Trypanosoma cruzi underlines the role of surface proteins during infection.
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DOI：
10.1371/journal.ppat.1006767
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发表时间：
2017-12
期刊：
影响因子：
6.7
通讯作者：
Teixeira SMR
中科院分区：
文献类型：
--
作者：
Belew AT;Junqueira C;Rodrigues-Luiz GF;Valente BM;Oliveira AER;Polidoro RB;Zuccherato LW;Bartholomeu DC;Schenkman S;Gazzinelli RT;Burleigh BA;El-Sayed NM;Teixeira SMR
克氏锥虫是引起恰加斯病的原生动物，具有复杂的生命周期，涉及几个形态和生物化学上不同的阶段，这些阶段与各种昆虫和哺乳动物宿主建立了复杂的相互作用。它还具有异质性种群结构，包括具有不同性质的菌株，如毒力、对药物的敏感性、抗原谱和组织趋向性。我们对两种克隆的克氏T. cruzi菌株进行了转录组比较分析，它们在动物感染模型中显示出不同的毒力表型：CL Brener是一个毒力克隆，CL-14是一个在体内感染模型中既没有感染性也没有致病性的克隆。对人类成纤维细胞感染后60和96小时收获的锥体和胞内锥体的基因表达分析显示，在感染哺乳动物细胞期间，寄生虫对不同环境的适应存在巨大差异，包括能量来源、氧化应激反应、细胞周期控制和细胞表面成分的变化。虽然在将这两种菌株的密乳线虫与60 hpi的密乳线虫进行比较时观察到广泛的转录组重塑，但在将96 hpi的密乳线虫与CL Brener的密乳线虫进行比较时，基因表达的差异要小得多。相反，从96 hpi无乳线虫向细胞外色乳线虫的分化与基因表达的显著变化有关，特别是编码表面蛋白的基因家族，如反式唾液酸酶、粘蛋白和粘蛋白相关表面蛋白（MASPs）。因此，我们的比较转录组分析表明，CL-14的无毒表型可能至少在一定程度上是由于编码表面蛋白的基因表达减少或延迟，这些基因与无乳线虫向色乳线虫的转变有关，这是在哺乳动物宿主中建立感染的重要步骤。证实了反式唾液酸酶家族成员在寄生虫分化中的作用，与野生型CL-14相比，转染后表达反式唾液酸酶基因的CL-14在感染细胞的上清液中显示出更快的trypmastigote释放动力学。恰加斯病由原生动物寄生虫克氏锥虫引起，是一种发生在若干拉丁美洲国家的感染，导致轻微疾病或对心脏和肠道造成严重损害。在恰加斯病患者中观察到的如此广泛的临床表现可能是由于宿主易感性的差异以及克鲁兹锥虫分离株之间存在很大的异质性。鉴定在寄生虫种群中差异表达的毒力因子是了解这种病原体与其宿主之间发生的独特相互作用的一种有价值的策略，这种相互作用可能导致发病，也可能不会导致发病。通过比较在感染动物模型中表现出不同毒力表型的两种克氏T.菌株的基因表达谱，我们确定了编码表面蛋白的基因的核心作用，该基因与细胞内复制性无尾线虫向感染性锥乳线虫的分化有关。我们发现这些基因的表达在两个菌株中发生差异，这种差异可能是影响寄生虫在哺乳动物宿主中生存和传播的一个因素。
Trypanosoma cruzi, the protozoan that causes Chagas disease, has a complex life cycle involving several morphologically and biochemically distinct stages that establish intricate interactions with various insect and mammalian hosts. It has also a heterogeneous population structure comprising strains with distinct properties such as virulence, sensitivity to drugs, antigenic profile and tissue tropism. We present a comparative transcriptome analysis of two cloned T. cruzi strains that display contrasting virulence phenotypes in animal models of infection: CL Brener is a virulent clone and CL-14 is a clone that is neither infective nor pathogenic in in vivo models of infection. Gene expression analysis of trypomastigotes and intracellular amastigotes harvested at 60 and 96 hours post-infection (hpi) of human fibroblasts revealed large differences that reflect the parasite’s adaptation to distinct environments during the infection of mammalian cells, including changes in energy sources, oxidative stress responses, cell cycle control and cell surface components. While extensive transcriptome remodeling was observed when trypomastigotes of both strains were compared to 60 hpi amastigotes, differences in gene expression were much less pronounced when 96 hpi amastigotes and trypomastigotes of CL Brener were compared. In contrast, the differentiation of the avirulent CL-14 from 96 hpi amastigotes to extracellular trypomastigotes was associated with considerable changes in gene expression, particularly in gene families encoding surface proteins such as trans-sialidases, mucins and the mucin associated surface proteins (MASPs). Thus, our comparative transcriptome analysis indicates that the avirulent phenotype of CL-14 may be due, at least in part, to a reduced or delayed expression of genes encoding surface proteins that are associated with the transition of amastigotes to trypomastigotes, an essential step in the establishment of the infection in the mammalian host. Confirming the role of members of the trans-sialidase family of surface proteins for parasite differentiation, transfected CL-14 constitutively expressing a trans-sialidase gene displayed faster kinetics of trypomastigote release in the supernatant of infected cells compared to wild type CL-14. Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is an infection that occurs in several Latin American countries, resulting in a mild illness or in severe damage of the heart and intestinal tract. Such a broad spectrum of clinical manifestations observed in Chagas disease patients is likely due to differences in host susceptibility as well as to a large heterogeneity among T. cruzi isolates. The identification of virulence factors that are differentially expressed in the parasite population is a valuable strategy for understanding of the distinct interactions that occur between this pathogen and its host, which may or may not lead to pathogenesis. By comparing the gene expression profiles of two T. cruzi strains that display contrasting virulence phenotypes in animal models of infection, we identified a central role for genes encoding surface proteins that is associated with the differentiation from intracellular replicative amastigotes to infective trypomastigotes. We showed that the expression of these genes occurs differentially within the two strains and this difference may be a factor that impacts parasite survival and dissemination in the mammalian host.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
2.1
作者：
Caro, Florence;Bercovich, Natalia;Vazquez, Martin P.
通讯作者：
Vazquez, Martin P.
影响因子：
6.7
作者：
Archer SK;Luu VD;de Queiroz RA;Brems S;Clayton C
通讯作者：
Clayton C
通过替代变量分析捕获基因表达研究中的异质性。
影响因子：
4.5
作者：
Leek, Jeffrey T.;Storey, John D.
通讯作者：
Storey, John D.
影响因子：
6.7
作者：
Erben ED;Fadda A;Lueong S;Hoheisel JD;Clayton C
通讯作者：
Clayton C