Transcriptome and Functional Genomics Reveal the Participation of Adenine Phosphoribosyltransferase in Trypanosoma cruzi Resistance to Benznidazole
Transcriptome and Functional Genomics Reveal the Participation of Adenine Phosphoribosyltransferase in Trypanosoma cruzi Resistance to Benznidazole
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DOI：
10.1002/jcb.25978
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Triana-Chavez, Omar
中科院分区：
文献类型：
--
作者：
Garcia-Huertas, Paola;Maria Mejia-Jaramillo, Ana;Triana-Chavez, Omar
目前，治疗克氏锥虫的唯一有效方法是苯硝唑(BZ)和硝呋莫司(NFX)。这种寄生虫对这些药物的作用机制和耐药性尚不完全清楚。为了鉴定敏感寄生虫和抗性寄生虫之间的差异表达转录本，我们对克氏锥虫转录组进行了大量的焦磷酸测序。此外，还分析了敏感和抗性寄生虫的2D凝胶电泳谱，并用功能基因组学来支持这些数据。结果表明，有133个基因在抗性寄生虫中有差异表达。转录组分析揭示了不同基因在不同功能和代谢途径上的调控，这可能表明克氏木霉的抗性是一个多基因过程。此外，利用转录组学方法，发现腺嘌呤磷酸核糖基转移酶(Aprt)基因在抗性寄生虫中表达下调，并通过双向电泳法证实了其表达谱。该基因在BZ抗性中的作用已被证实，它在敏感和抗性寄生虫中过表达。有趣的是，这两种寄生虫对BZ和过氧化氢都变得更加敏感。这是首次对克氏锥虫中与BZ抗性相关的调控基因进行RNA-SEQ研究，并揭示aprt在克氏锥虫抗药性中的作用。虽然克氏木霉的调控主要是转录后调控，但转录组分析在2D凝胶分析和功能基因组学的支持下，提供了抗性条件下基因表达谱的总体思路。这些结果为进一步了解克氏毛滴虫对BZ的作用和抗性机制提供了必要的信息。(C)2017威利期刊公司。
Currently, the only available treatments for Trypanosoma cruzi are benznidazole (Bz) and nifurtimox (Nfx). The mechanisms of action and resistance to these drugs in this parasite are not complete known. In order to identify differentially expressed transcripts between sensitive and resistant parasites, a massive pyrosequencing of the T. cruzi transcriptome was carried out. Additionally, the 2D gel electrophoresis profile of sensitive and resistant parasites was analyzed and the data were supported with functional genomics. The results showed 133 differentially expressed genes in resistant parasites. The transcriptome analysis revealed the regulation of different genes with several functions and metabolic pathways, which could suggest that resistance in T. cruzi is a multigenic process. Additionally, using transcriptomics, one gene, adenine phosphoribosyltransferase (APRT), was found to be down-regulated in the resistant parasites and its expression profile was confirmed by 2D electrophoresis analysis. The role of this gene in the resistance to Bz was confirmed overexpressing it in sensitive and resistant parasites. Interestingly, both parasites became more sensitive to Bz and H2O2. This is the first RNA-seq study to identify regulated genes in T. cruzi associated with Bz resistance and to show the role of APRT in T. cruzi resistance. Although T. cruzi regulation is mainly post-transcriptional, the transcriptome analysis, supported by 2D gel analysis and functional genomic, provides an overall idea of the expression profiles of genes under resistance conditions. These results contribute essential information to further the understanding of the mechanisms of action and resistance to Bz in T. cruzi. (C) 2017 Wiley Periodicals, Inc.