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Adverse Health Effects of Cylindrospermopsin in Laboratory Animal (Mouse) Studies in Multi-day Study using Affymetrix Array

ABSTRACT: The goal of this project was to determine gene expression changes in the liver at different times post treatment with the common freshwater cyanobacterial toxin, cylindrospermopsin (CYN). The liver is generally considered to be the primary target of this toxin and all the data has been derived from livers of exposed animals as well as concurrent controls. Preliminary gene expression effects have been reported (Chernoff et al., 2010) and the current data extend the time course from 1 hour post exposure to 1-day post cessation of a 5-day period dosing period. Gene expression changes were determined in the livers of pregnant mice exposed to toxic levels of the cyanobacterial toxin, cylindrospermopsin. Animals were exposed to a dose of CYN that elicited significant hepatic toxicity. This multi-day study used tissue obtained 1-day, 2-days, 4-days, and 5-days after the initiation of a 5 consecutive day dosing regimen. Toxicity data including gross observations, necropsy findings and clinical chemistry endpoints indicative of hepatic, renal and homeostasis functions were collected at the same time liver tissues for gene expression analysis were obtained. Gene expression changes were analyzed on the GeneChipÂ® Mouse Genome 430 2.0 Array.

ORGANISM(S): Mus musculus

SUBMITTER: Neil Chernoff

PROVIDER: E-GEOD-83198 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Adverse Health Effects of Cylindrospermopsin in Laboratory Animal (Mouse) Studies in Single Day Study using Affymetrix Array

Project description:The goal of this project was to determine gene expression changes in the liver at different times post treatment with the common freshwater cyanobacterial toxin, cylindrospermopsin (CYN). The liver is generally considered to be the primary target of this toxin and all the data has been derived from livers of exposed animals as well as concurrent controls. Preliminary gene expression effects have been reported (Chernoff et al., 2010) and the current data extend the time course from 1 hour post exposure to 1-day post cessation of a 5-day period dosing period. Gene expression changes were determined in the livers of pregnant mice exposed to toxic levels of cyanobacterial toxin, cylindrospermopsin. Animals were exposed to a dose of CYN that elicited significant hepatic toxicity. This single dose study analyzed liver samples collected at 1, 3, 6, 12 and 24 hours post dose. Gene expression changes were analyzed on the GeneChipÂ® Mouse Genome 430 2.0 Array.

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Identification of key pathways involved in the toxic response of the cyanobacterial toxin cylindrospermopsin on human hepatic HepaRG cells

Project description:Cylindrospermopsin (CYN), a hepatotoxin produced by several cyanobacterial species, has been implied in human intoxications following ingestion. Although some pathways involved in CYN toxicity have been reported, the whole process is certainly more complex. In order to identify the mechanisms related to CYN toxicity on liver, we studied the transcriptomic profile of HepaRG human hepatic cells exposed to a low concentration of CYN (0.8 µM for 24 hrs). The affected pathways were further confirmed through the expression of key genes by qPCR, and the investigation of toxicity markers by high content analysis (HCA) on a broader CYN concentration range. Also CYP450 activities and cell redox homeostasis were investigated after acute and repeated exposure to CYN. Our results revealed that CYN induced up-regulation of genes implied in RNA modification and maturation, but also down-regulation of genes involved in xenobiotic metabolism and cell cycle progress. There was cell cycle disturbance characterised by an accumulation of G1/S and G2/M cells, and an increase of phospho-H3 positive cells. The cell cycle arrest was linked to the induction of DNA damage pointed out by an increase of γH2AX positive cells, but also an accumulation of sub G1 cells indicating apoptosis not involving caspase 3. While GSH content decreased strongly after acute exposure to CYN, it increased after a repeated exposure reflecting an adaptive response of cell redox homeostasis when exposure to CYN was prolonged. However, our data suggested also that CYN induced a down-regulation of phase I and II metabolism gene products, and CYP450 activities were affected following both acute and repeated exposure to CYN. Our study indicated that repeated exposure of liver cells to low concentrations of CYN may affect their detoxification capacities.

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Modulation of gene expression induced by cylindrospermopsin in human intestinal Caco-2 cells

Project description:Cylindrospermopsin (CYN) is a cyanotoxin that has been recognised as an emerging potential public health risk. Although CYN toxicity has been demonstrated, the mechanisms involved have not been fully characterised. To identify some key pathways related to this toxicity, we studied the transcriptomic profile of human intestinal Caco-2 cells exposed to a sub-toxic concentration of CYN (1.6 M-BM-5M for 24hrs) using a non-targeted approach. CYN was shown to modulate different biological functions which were related to growth arrest (with down-regulation of cdkn1a and uhrf1 genes), and DNA recombination and repair (with up-regulation of aptx and pms2 genes). Our main results reported an increased expression of some histone-modifying enzymes (histone acetyl and methyltransferases MYST1, KAT5 and EHMT2) involved in chromatin remodelling, which is essential for initiating transcription. We also detected greater levels of acetylated histone H2A (Lys5) and dimethylated histone H3 (Lys4), two products of these enzymes. In conclusion, CYN overexpressed proteins involved in DNA damage repair and transcription, including modifications of nucleosomal histones. Our results highlighted some new cell processes induced by CYN. Modulation of gene expression in differentiated Caco-2 cells was measured at 24 hours after exposure to doses of 0 and 1.6 M-BM-5M of cylindrospermopsin. Six independent experiments were performed, but only 11 samples (5 controls and 6 exposed to cylindrospermopsin) had an RNA quality indicator M-bM-^IM-% 9 and were used for the microarray experiments.

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Effect of Microcystin-LR and cylindrospermopsin (CYN) on gene expression in human hepatocytes, HepaRG cells and HCC HepG2 cells.

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