Transcriptome Remodeling in Trypanosoma cruzi and Human Cells during Intracellular Infection.
Transcriptome Remodeling in Trypanosoma cruzi and Human Cells during Intracellular Infection.
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DOI：
10.1371/journal.ppat.1005511
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发表时间：
2016-04
期刊：
影响因子：
6.7
通讯作者：
Burleigh BA
中科院分区：
文献类型：
--
作者：
Li Y;Shah-Simpson S;Okrah K;Belew AT;Choi J;Caradonna KL;Padmanabhan P;Ndegwa DM;Temanni MR;Corrada Bravo H;El-Sayed NM;Burleigh BA
细胞内定植和持续感染的动质体原生动物寄生虫，克氏锥虫，人类恰加斯病的发病机制的基础。为了获得对T.在Cruzi感染过程中，在人成纤维细胞的感染时间过程中，在寄生虫和宿主细胞中同时捕获转录组动态。大规模改造的T。在细胞内感染的早期建立期间观察到Cruzi转录组，与寄生虫的主要发育转变一致。与这种早期反应相反，一旦T细胞成熟，在稳态mRNA水平上几乎没有检测到额外的变化。形成克氏无鞭毛体。我们的研究结果表明，转录组重塑需要建立一个修改的模板，以指导寄生虫的发育过渡，而自我平衡功能的调节独立的转录组的变化，类似于相关的锥虫报道。尽管T. cruzi，来自不同发育阶段的转录组签名反映了T.克鲁兹生物学聚焦于能量代谢，我们能够验证mRNA表达谱中的预测。我们证明了T的不同感染阶段的生物能量特性的可测量的差异。cruzi的研究，并提出了额外的发现，强调线粒体电子传递的重要性，在T。cruzi无鞭毛体的生长和存活。T的后果。宿主的克氏定殖包括免疫应答基因和细胞周期调节因子的动态表达，以及宿主胆固醇和脂质合成途径的上调，这可能有助于刺激细胞内T细胞。克鲁奇生长因此，除了从寄生虫和宿主中差异表达基因的基因本体论和功能富集分析获得的生物学推断之外，我们的全面的高分辨率转录组数据集提供了对T. Cruzi感染生物学，并为未来的药物和疫苗发现工作提供了基础。深入了解宿主定植和病原微生物传播的功能过程，对于推进有效的干预战略至关重要。这项研究的重点是克氏锥虫，媒介传播的原生动物寄生虫负责人类恰加斯病和全球感染性心肌炎的主要原因。为了全面了解这种寄生虫的生物学及其与哺乳动物宿主细胞的相互作用，我们利用深度测序方法生成了T哺乳动物感染阶段的全面、高分辨率转录组图谱。cruzi与跨感染时程的人类宿主细胞转录组的同时询问。我们证明了细胞内T.哺乳动物宿主细胞中的Cruzi感染伴随着寄生虫和宿主细胞转录组的广泛重塑。尽管锥虫缺乏转录控制机制，我们的分析确定了在T。与寄生虫的已知或预测的生物学特征一致的cruzi。细胞内T细胞生物学的新见解。本研究中获得的cruzi感染和无鞭毛体发育调控为功能网络分析奠定了独特的基础，这将有助于提供寄生虫依赖性和宿主功能途径之间的功能联系，这些途径有可能用于干预。
Intracellular colonization and persistent infection by the kinetoplastid protozoan parasite, Trypanosoma cruzi, underlie the pathogenesis of human Chagas disease. To obtain global insights into the T. cruzi infective process, transcriptome dynamics were simultaneously captured in the parasite and host cells in an infection time course of human fibroblasts. Extensive remodeling of the T. cruzi transcriptome was observed during the early establishment of intracellular infection, coincident with a major developmental transition in the parasite. Contrasting this early response, few additional changes in steady state mRNA levels were detected once mature T. cruzi amastigotes were formed. Our findings suggest that transcriptome remodeling is required to establish a modified template to guide developmental transitions in the parasite, whereas homeostatic functions are regulated independently of transcriptomic changes, similar to that reported in related trypanosomatids. Despite complex mechanisms for regulation of phenotypic expression in T. cruzi, transcriptomic signatures derived from distinct developmental stages mirror known or projected characteristics of T. cruzi biology. Focusing on energy metabolism, we were able to validate predictions forecast in the mRNA expression profiles. We demonstrate measurable differences in the bioenergetic properties of the different mammalian-infective stages of T. cruzi and present additional findings that underscore the importance of mitochondrial electron transport in T. cruzi amastigote growth and survival. Consequences of T. cruzi colonization for the host include dynamic expression of immune response genes and cell cycle regulators with upregulation of host cholesterol and lipid synthesis pathways, which may serve to fuel intracellular T. cruzi growth. Thus, in addition to the biological inferences gained from gene ontology and functional enrichment analysis of differentially expressed genes in parasite and host, our comprehensive, high resolution transcriptomic dataset provides a substantially more detailed interpretation of T. cruzi infection biology and offers a basis for future drug and vaccine discovery efforts. In-depth knowledge of the functional processes governing host colonization and transmission of pathogenic microorganisms is essential for the advancement of effective intervention strategies. This study focuses on Trypanosoma cruzi, the vector-borne protozoan parasite responsible for human Chagas disease and the leading cause of infectious myocarditis worldwide. To gain global insights into the biology of this parasite and its interaction with mammalian host cells, we have exploited a deep-sequencing approach to generate comprehensive, high-resolution transcriptomic maps for mammalian-infective stages of T. cruzi with the simultaneous interrogation of the human host cell transcriptome across an infection time course. We demonstrate that the establishment of intracellular T. cruzi infection in mammalian host cells is accompanied by extensive remodeling of the parasite and host cell transcriptomes. Despite the lack of transcriptional control mechanisms in trypanosomatids, our analyses identified functionally-enriched processes within sets of developmentally-regulated transcripts in T. cruzi that align with known or predicted biological features of the parasite. The novel insights into the biology of intracellular T. cruzi infection and the regulation of amastigote development gained in this study establish a unique foundation for functional network analyses that will be instrumental in providing functional links between parasite dependencies and host functional pathways that have the potential to be exploited for intervention.