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Transcriptomic alterations in a myoblast cell line infected with four distinct strains of Trypanosoma cruzi

ABSTRACT: We examined the extent to which different Trypanosoma cruzi strains induce transcriptomic changes in cultured L6E9 myoblasts 72 hours or 48 hours after infection with four strains of the parasite [Brazil (TCI); Y, CL and Tulahuen (TC II) strains]. Expression of 6,289 distinct fully annotated unigenes was quantified with 27k rat oligonucleotide arrays in each of the four replicas of all control and infected RNA samples. Considering changes >1.5-fold and p-val<0.05, the Tulahuen strain was the most disruptive to host transcriptome, with (17% significantly altered genes), while in Y strain altered only 6% of the genes were altered. The significantly altered genes in the infected cells were largely different among the four strains, with only 21 genes changed in the same direction being similarly changed by all four strains. However, when expression ratios of all genes were compared, myoblasts infected with different strains showed proportional overall genbe expression alterations. These results indicate that infection with different parasite strains modulates similar but not identical pathways in the host cells.

ORGANISM(S): Rattus norvegicus

PROVIDER: GSE18175 | GEO | 2009/09/23

SECONDARY ACCESSION(S): PRJNA119533

REPOSITORIES: GEO

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Transcriptomic alterations in a myoblast cell line infected with four distinct strains of Trypanosoma cruzi

Project description:We examined the extent to which different Trypanosoma cruzi strains induce transcriptomic changes in cultured L6E9 myoblasts 72 hours or 48 hours after infection with four strains of the parasite [Brazil (TCI); Y, CL and Tulahuen (TC II) strains]. Expression of 6,289 distinct fully annotated unigenes was quantified with 27k rat oligonucleotide arrays in each of the four replicas of all control and infected RNA samples. Considering changes >1.5-fold and p-val<0.05, the Tulahuen strain was the most disruptive to host transcriptome, with (17% significantly altered genes), while in Y strain altered only 6% of the genes were altered. The significantly altered genes in the infected cells were largely different among the four strains, with only 21 genes changed in the same direction being similarly changed by all four strains. However, when expression ratios of all genes were compared, myoblasts infected with different strains showed proportional overall genbe expression alterations. These results indicate that infection with different parasite strains modulates similar but not identical pathways in the host cells. 20 µg total RNA extracted in Trizol from each of the 20 cell culture dishes containing control or infected with (Y, CL, Tulahuen or Brazil) Trypanosoma cruzi strains (4 dishes for each strain) was reverse transcribed in the presence of fluorescent Alexa Fluor®_647 or Alexa Fluor®_555-aha-dUTPs (Invitrogen, CA) to obtain labeled cDNAs. Red and green-labeled samples of biological replicas were then co-hybridized (“multiple yellow” strategy) overnight at 50°C with Operon v3.0 Rat 27k oligonucleotide arrays printed by Duke University (GPL9207). After washing (0.1% SDS and 1% SSC) to remove the non-hybridized cDNA, each array was scanned with GenePix 4000B scanner (MDS, Toronto, Canada) and images were primarily analyzed with GenePixPro 6.0 (Axon Instruments, CA).

2009-09-22 | E-GEOD-18175 | biostudies-arrayexpress

Different genotypes of Trypanosoma cruzi produce distinctive placental environment genetic response in chronic experimental infection

Project description:Congenital infection of Trypanosoma cruzi allows transmission of this parasite through generations. Despite the problematic that this entails, little is known about the placenta environment genetic response produced against infection. We performed functional genomics by microarray analysis in C57Bl/6J mice comparing placentas from uninfected animals and from animals infected with two different T. cruzi strains: K98, a clone of the non-lethal myotropic CA-I strain (TcI), and VD (TcVI), isolated from a human case of congenital infection. Analysis of networks by GeneMANIA of differentially expressed genes showed that “Secretory Granule” was a pathway down-regulated in both infected groups, whereas “Innate Immune Response” and “Response to Interferon-gamma” were pathways up-regulated in VD infection but not in K98. Applying another approach, the GSEA algorithm that detects small changes in predetermined gene sets, we found that metabolic processes, transcription and macromolecular transport were down-regulated in infected placentas environment and some pathways related to cascade signaling had opposite regulation: over-represented in VD and down-regulated in K98 group. We also have found a stronger placental tropism by VD strain, by detection of parasite DNA and RNA, suggesting living parasites in that tissue. Our study is the first one to describe in a murine model the genetic response of placental environment to T. cruzi infection and suggests the development of a strong immune response, parasite genotype-dependent, to the detriment of cellular metabolism.

2016-08-25 | GSE85996 | GEO

Different genotypes of Trypanosoma cruzi produce distinctive placental environment genetic response in chronic experimental infection

Project description:Congenital infection of Trypanosoma cruzi allows transmission of this parasite through generations. Despite the problematic that this entails, little is known about the placenta environment genetic response produced against infection. We performed functional genomics by microarray analysis in C57Bl/6J mice comparing placentas from uninfected animals and from animals infected with two different T. cruzi strains: K98, a clone of the non-lethal myotropic CA-I strain (TcI), and VD (TcVI), isolated from a human case of congenital infection. Analysis of networks by GeneMANIA of differentially expressed genes showed that âSecretory Granuleâ was a pathway down-regulated in both infected groups, whereas âInnate Immune Responseâ and âResponse to Interferon-gammaâ were pathways up-regulated in VD infection but not in K98. Applying another approach, the GSEA algorithm that detects small changes in predetermined gene sets, we found that metabolic processes, transcription and macromolecular transport were down-regulated in infected placentas environment and some pathways related to cascade signaling had opposite regulation: over-represented in VD and down-regulated in K98 group. We also have found a stronger placental tropism by VD strain, by detection of parasite DNA and RNA, suggesting living parasites in that tissue. Our study is the first one to describe in a murine model the genetic response of placental environment to T. cruzi infection and suggests the development of a strong immune response, parasite genotype-dependent, to the detriment of cellular metabolism. Total RNA obtained from isolated placentas from mice with chronic T. cruzi infection (K98 or VD strains) at 18.5 days of gestation compared to uninfected control mice.

2016-08-25 | E-GEOD-85996 | biostudies-arrayexpress

Systematic detection of host pathways universally inhibited by Plasmodium yoelii parasites for immune intervention

Project description:Malaria is a disease with diverse symptoms depending on host immune status and pathogenicity of Plasmodium parasites. The continuous parasite growth within a host suggests mechanisms of immune evasion and/or inhibition. To identify pathways commonly inhibited by malaria infection, we infected C67BL/6 mice with four Plasmodium yoelii strains causing different disease phenotypes and 24 progeny of a genetic cross. mRNAs from mouse spleens day 1 and/or day 4 post infection (p.i.) were hybridized to a mouse microarray to identify activated or inhibited pathways, upstream regulators, and linkages to parasite genetic loci. Strong interferon responses were observed after infection with N67 strain, whereas initial inhibition and later activation of hematopoiesis pathways were found after infection with 17XNL parasite. Inhibition of pathways such as Th1 activation, dendritic cell (DC) maturation, and NFAT immune regulation were observed in mice infected with all the parasite strains day 4 p.i., suggesting universally inhibited immune pathways. Treatment of infected mice with antibodies against T cell receptors OX40 or CD28 to activate malaria-inhibited pathways enhanced host survival. Controlled activation of these pathways may provide important strategies for better disease management and for developing an effective vaccine.

2018-10-29 | GSE114718 | GEO

Transcriptomic analysis of Plasmodium PBANKA, PBSLTRiP-KO, PB268-KO parasite infected and uninfected host cell

2016-01-01 | GSE53292 | GEO

Microarray Expression Analysis of 3 Canonical Toxoplasma infections of human neuroepithelial cells

Project description:The outcome of infections with Toxoplasma gondii in humans is dependent in part on the genetic makeup of the infecting organism. Recent studies have indicated that most infecting Toxoplasma organisms fall into 1 of 3 canonical lineages. Previous studies have investigated the effects of Toxoplasma on its host cell transcriptome. Little is known, however, about the effects of three canonical lineages on brain cells, the principal site of parasite lifelong persistence. In this study, we examined the transcriptional profile of human neuroepithelioma cells in response to T. gondii infection using microarray analysis to characterize the strain-specific host cell response to 3 canonical T. gondii strains. We found that the extent of the expression changes varied considerably among the three strains. Neuroepithelial cells infected with type I exhibited the most differential gene expression, whereas type II infected cells had a substantially smaller number of genes which were differentially expressed. Cells infected with type III exhibited intermediate effects on gene expression. The three strains also differed in the individual genes and gene pathways which were altered following cellular infection. For example, gene ontology (GO) analysis indicated that type I infection largely affects genes related to central nervous system while type III infection largely alters genes which affect nucleotide metabolism; type II infection does not alter expression of a clearly defined set of genes. Moreover, Ingenuity pathway analysis (IPA) revealed the sophistication of different strain in its interactions with the host. These differences may explain some of the variation in the neurobiological effects of different strains of Toxoplasma on infected individuals.

2011-06-24 | GSE22986 | GEO

Comparative proteomic of Trypanosoma cruzi strains

Project description:Trypanosoma cruzi, the causative agent of Chagas disease, is a genetically diverse with TcI being the most widespread lineage in the Americas. The protein expression dynamics allows unravel the biological phenomena of pathogens such as disease development and evolution. A large number of studies have shown a direct relationship between genetic variability and the biological characteristics of T. cruzi. However, much remains to be explored regarding the impact of gene expression on the development and evolution of Chagas disease. Two-dimensional (2D) electrophoresis and mass-spectrometry were employed to characterize the global protein expression patterns of epimastigotes from four distinct TcI strains, which display different growth kinetics. Biological differences were identified and confirmed throug metabolic and morphometric assays. Statistical tests (parametric or non-parametric test, as appropriate for the experiment) were used to assess the significance of the observed differences. The Ascending hierarchical classification analysis shows two clusters, that were congruent with their growth kinetics. The proteins expressed in one or the other cluster were identified by mass spectrometry. These proteins were evaluated by independent biological assays, which showed that the parasite’s metabolic activity and morphology is significantly different between the two clusters. We conclude that the protein expression patterns displayed by epimastigotes from fast/slow growth TcI strains are correlated with various biological parameters, including intensity of metabolic activity, metabolic fuel usage and resistance to oxidative stress, as well as morphological characteristics, such as length of the flagellum.

2023-07-12 | PXD032748 | Pride

Transcriptome analysis of mouse hearts infected with Trypanosoma cruzi: Differential modulation of gene expression by two strains

Project description:Purpose: One important challenge in the field of Chagas disease research is to determine the main factors leading to the establishment of the parasite in some organs in the chronic stage, mainly the heart and/or digestive system. Our group previously showed that distinct strains of T. cruzi (JG and Col1.7G2) presented differential tissue distribution in the chronic stage in double-infected BALB/c mice. To investigate changes in the host triggered by the two distinct T. cruzi strains, we assessed the gene expression profile of BALB/c mouse hearts infected with either JG, Col1.7G2 or an equivalent mixture of both parasites during the initial phase of infection. Results: We demonstrate a clear distinction in host gene expression modulation by both parasites. Col1.7G2 strongly activated Th1-polarized immune response genes, whereas JG showed minor immune response activation. Moreover, JG strongly impaired the expression of genes for ribosomal proteins and mitochondrial proteins related to the electron transport chain and the citric acid cycle. Interestingly, evaluation of gene expression in mice inoculated with the mixture of parasites showed expression profiles for both up- and downregulated genes, indicating the coexistence of both parasite strains in the heart during the acute phase. Conclusions: We suggest that different strains of T. cruzi may be distinguished by their efficiency in activating the immune system, modulating host energy and reactive oxygen species production and impairing protein synthesis during early infection, which may be decisive in defining parasite persistence in specific organs.

2020-08-19 | GSE132132 | GEO

Transcriptomic analysis of Plasmodium PBANKA, PBSLTRiP-KO, PB268-KO parasite infected and uninfected host cell

Project description:Liver stage of malaria parasite exports SLTRiP and PB268 to the cytosol of parasite infected host cell. To know the host genes perturbed by WT-PBANKA, SLTRiP-KO and PB268-KO parasite growth, we did transcriptomic sequencing of infected host cells. We did mRNA sequencing of four samples for comparative analysis of WT and PB-knockout parasites infected host cells at 22 hours of post sporozoites infection. mRNA profiles of Plasmodium PBANKA, PBSLTRiP-KO, PB268-KO parasite infected and uninfected HepG2 cells after 22hrs of sporozoites infections were generated by deep sequencing using Illumina GAIIx.

2016-07-03 | E-GEOD-53292 | biostudies-arrayexpress

The redox homeostasis of skeletal muscle cells regulates stage differentiation of Toxoplasma gondii

Project description:Toxoplasma gondii is an obligatory intracellular parasite that causes persistent infections in birds and mammals including ~30% of the world’s human population. Differentiation from proliferative and metabolically active tachyzoites to largely dormant bradyzoites initiates the chronic phase of infection and occurs predominantly in brain and muscle tissues. Here we used murine skeletal muscle cells (SkMCs) to decipher host cellular factors that favor T. gondii bradyzoite formation in terminally differentiated and syncytial myotubes, but not in proliferating myoblast precursors. Genome-wide transcriptome analyses of T. gondii-infected SkMCs and non-infected controls identified ~6,500 genes which were differentially expressed (DEGs) in myotubes compared to myoblasts, largely irrespective of infection. On the other hand, genes related to central carbohydrate metabolism, to redox homeostasis, and to the Nrf2-dependent stress response pathway were enriched in both infected myoblast precursors and myotubes. Stable isotope-resolved metabolite profiling indicated increased fluxes into the oxidative branch of the pentose phosphate pathway (OxPPP) in infected myoblasts and into the TCA cycle in infected myotubes. High OxPPP activity in infected myoblasts was associated with increased NADPH/NADP+ ratio while myotubes exhibited higher ROS levels and lower expression of anti-oxidants and detoxification enzymes. Pharmacological reduction of ROS levels in SkMCs inhibited bradyzoite differentiation, while increased ROS induced bradyzoite formation. Thus, we identified a novel host cell-dependent mechanism that triggers stage conversion of T. gondii into persistent tissue cysts in its natural host cell type.

2021-11-04 | GSE133952 | GEO

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