Metallome of cerebrovascular endothelial cells infected with <i>Toxoplasma gondii</i> using μ-XRF imaging and inductively coupled plasma mass spectrometry

Al-sandaqchi, Alaa; Brignell, Chris; Collingwood, Joanna F.; Geraki, Kalotina; Mirkes, Evgeny M.; Kong, Kenny; Castellanos, Marcos; May, Sean; Stevenson, Carl W.; Elsheikha, Hany M.

doi:10.1039/c8mt00136g

Cited by 15 publications

(17 citation statements)

References 42 publications

(40 reference statements)

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“…This finding is consistent with previous transcriptome studies in murine macrophages [49] and human fibroblast cells [50], reaffirming the deregulation of the immune response following T. gondii infection of hBMECs. The increased expression of the MT1M gene was also reported previously [51], where MT1M upregulation has been suggested to contribute to the cellular homeostasis of transition metals, particularly zinc, during T. gondii infection. The down-regulation of microRNA 29a has also been reported in a mouse brain infected by the cystogenic strain (type II) [52], which is less virulent than the RH (type I) strain used in our study, suggesting that inhibition of microRNA 29a is common in T. gondii infection, independent of the parasite virulence.…”

Section: Discussionsupporting

confidence: 82%

Transcriptome Profiling of Toxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin

et al. 2020

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Central to the progression of cerebral toxoplasmosis is the interaction of Toxoplasma gondii with the blood-brain barrier (BBB) endothelial cells. In the present work, we tested the hypothesis that inhibition of Wnt pathway signalling by the monovalent ionophore monensin reduces the growth of T. gondii infecting human brain microvascular endothelial cells (hBMECs) or microglial cells. The anti-parasitic effect of monensin (a Wnt signalling inhibitor) on the in vitro growth of T. gondii tachyzoites was investigated using two methods (Sulforhodamine B staining and microscopic parasite counting). The monensin inhibited T. gondii growth (50% inhibitory concentration [IC50] = 0.61 μM) with a selective index = 8.48 when tested against hBMECs (50% cytotoxic concentration [CC50] = 5.17 μM). However, IC50 of monensin was 4.13 μM with a SI = 13.82 when tested against microglia cells (CC50 = 57.08 μM), suggesting less sensitivity of microglia cells to monensin treatment. The effect of T. gondii on the integrity of the BBB was assessed by the transendothelial electrical resistance (TEER) assay using an in vitro human BBB model. The results showed that T. gondii infection significantly decreased hBMECs’ TEER resistance, which was rescued when cells were treated with 0.1 µM monensin, probably due to the anti-parasitic activity of monensin. We also investigated the host-targeted effects of 0.1 µM monensin on global gene expression in hBMECs with or without T. gondii infection. Treatment of hBMECs with monensin did not significantly influence the expression of genes involved in the Wnt signalling pathway, suggesting that although inhibition of the Wnt signalling pathway did not play a significant role in T. gondii infection of hBMECs, monensin was still effective in limiting the growth of T. gondii. On the contrary, monensin treatment downregulated pathways related to steroids, cholesterol and protein biosynthesis and their transport between endoplasmic reticulum and Golgi apparatus, and deregulated pathways related to cell cycle and DNA synthesis and repair mechanisms. These results provide new insight into the host-modulatory effect of monensin during T. gondii infection, which merits further investigation.

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Section: Discussionsupporting

confidence: 82%

Transcriptome Profiling of Toxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin

et al. 2020

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“…Recent studies show that T. gondii infection can induce significant structural, functional and metabolic changes in the brain microvascular endotehlial cells (Al-Sandaqchi et al, 2018;Hu et al, 2018;Ma et al, 2019;Al-Sandaqchi et al, 2020;Harun et al, 2020a;Harun et al, 2020b) and can change the neuron subpopulations (Odorizzi et al, 2010). However, the exact mechanisms of behavioral abnormalities and change in the subpopulations of neurons induced by T. gondii infection remains to be clearly defined.…”

Section: Rop18-mediated Transcriptional Reprogramming Of Hek293t Cellsmentioning

confidence: 99%

ROP18-Mediated Transcriptional Reprogramming of HEK293T Cell Reveals New Roles of ROP18 in the Interplay Between Toxoplasma gondii and the Host Cell

Elsheikha

et al. 2020

Front. Cell. Infect. Microbiol.

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Toxoplasma gondii secretes a number of virulence-related effector proteins, such as the rhoptry protein 18 (ROP18). To further broaden our understanding of the molecular functions of ROP18, we examined the transcriptional response of human embryonic kidney cells (HEK293T) to ROP18 of type I T. gondii RH strain. Using RNA-sequencing, we compared the transcriptome of ROP18-expressing HEK293T cells to control HEK293T cells. Our analysis revealed that ROP18 altered the expression of 750 genes (467 upregulated genes and 283 downregulated genes) in HEK293T cells. Gene ontology (GO) and pathway enrichment analyses showed that differentially expressed genes (DEGs) were significantly enriched in extracellular matrix– and immune–related GO terms and pathways. KEGG pathway enrichment analysis revealed that DEGs were involved in several disease-related pathways, such as nervous system diseases and eye disease. ROP18 significantly increased the alternative splicing pattern “retained intron” and altered the expression of 144 transcription factors (TFs). These results provide new insight into how ROP18 may influence biological processes in the host cells via altering the expression of genes, TFs, and pathways. More in vitro and in vivo studies are required to substantiate these findings.

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“…Likewise, the use of spectroscopic techniques, such as Fourier Transform Infrared (FTIR), can reveal alterations in the chemical constituents of infected cells [ 14 ]. Additionally, synchrotron radiation X-ray fluorescence (μSR-XRF) is a chemical element imaging technique that can be used to generate X-ray fluorescence elemental maps of biological tissues [ 15 , 16 , 17 ], with detection sensitivity and spatial resolution well-suited to characterize host–parasite interactions. Together, transcriptomics, spectroscopic FTIR and μSR-XRF approaches can provide an integrated global view of the transcriptional, chemical, and elemental changes that occur in BBB endothelial cells in response to N. caninum infection.…”

Section: Introductionmentioning

confidence: 99%

Impact of Neospora caninum Infection on the Bioenergetics and Transcriptome of Cerebrovascular Endothelial Cells

et al. 2020

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In this work, the effects of the protozoan Neospora caninum on the bioenergetics, chemical composition, and elemental content of human brain microvascular endothelial cells (hBMECs) were investigated. We showed that N. caninum can impair cell mitochondrial (Mt) function and causes an arrest in host cell cycling at S and G2 phases. These adverse effects were also associated with altered expression of genes involved in Mt energy metabolism, suggesting Mt dysfunction caused by N. caninum infection. Fourier Transform Infrared (FTIR) spectroscopy analysis of hBMECs revealed alterations in the FTIR bands as a function of infection, where infected cells showed alterations in the absorption bands of lipid (2924 cm−1), amide I protein (1649 cm−1), amide II protein (1537 cm−1), nucleic acids and carbohydrates (1092 cm−1, 1047 cm−1, and 939 cm−1). By using quantitative synchrotron radiation X-ray fluorescence (μSR-XRF) imaging and quantification of the trace elements Zn, Cu and Fe, we detected an increase in the levels of Zn and Cu from 3 to 24 h post infection (hpi) in infected cells compared to control cells, but there were no changes in the level of Fe. We also used Affymetrix array technology to investigate the global alteration in gene expression of hBMECs and rat brain microvascular endothelial cells (rBMVECs) in response to N. caninum infection at 24 hpi. The result of transcriptome profiling identified differentially expressed genes involved mainly in immune response, lipid metabolism and apoptosis. These data further our understanding of the molecular events that shape the interaction between N. caninum and blood-brain-barrier endothelial cells.

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Metallome of cerebrovascular endothelial cells infected with Toxoplasma gondii using μ-XRF imaging and inductively coupled plasma mass spectrometry

Cited by 15 publications

References 42 publications

Transcriptome Profiling of Toxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin

Transcriptome Profiling of Toxoplasma gondii-Infected Human Cerebromicrovascular Endothelial Cell Response to Treatment with Monensin

ROP18-Mediated Transcriptional Reprogramming of HEK293T Cell Reveals New Roles of ROP18 in the Interplay Between Toxoplasma gondii and the Host Cell

Impact of Neospora caninum Infection on the Bioenergetics and Transcriptome of Cerebrovascular Endothelial Cells

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