Unknown,Transcriptomics,Genomics,Proteomics

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Microarray characterization of gene expression changes in blood during acute ethanol exposure

ABSTRACT: As part of the civil aviation safety program to define the adverse effects of ethanol on flying performance, we present results of our DNA microarray analysis of samples from a timecourse study of individuals given ethanol orally, and then evaluated by breathalyzer to monitor blood alcohol content (BAC). At five blood alcohol levels, T1-T5, blood was drawn such that the samples represented 0%, 0.04%, 0.08% BAC, and return to 0.04%, and 0.02% BAC. Microarray analysis showed that changes in gene expression could be detected across the time-course. We verified these expression changes by quantitative polymerase chain reaction (qPCR). Candidate target genes identified from the microarray analysis were clustered by expression change pattern, examined for shared functions and functional network membership. Five coordinately expressed groups were revealed and functional analysis showed shared transcription factor binding sites and functions for members of the clusters. These functions include protein synthesis and modification, expected for changes in gene expression, hematological and immune functions, expected for a blood sample, and pancreatic and hepatic function, expected as response to ethanol. The results provide a first look at changing gene expression patterns in blood during acute increase of ethanol concentration and its depletion due to metabolism or excretion and demonstrate that it is possible to detect significant changes in gene expression using total RNA isolated from whole blood. The analysis approach for this study can be utilized as part of a workflow to identify target genes by timecourse changes in gene expression that may affect pilot performance. Blood samples were collected from six male subjects given a mixture of orange and vodka such that their BAC would reach a maximum level of 0.08%. Five blood samples, T1-5, were taken from each subject, 0% prior to drinking alcohol, 0.04% and 0.08% as their BAC increased and 0.04% and 0.02% BAC as the alcohol concentration declined. Blood samples were taken from a control set of five male subjects drinking only orange juice. Five sample collections at times averaged from the ethanol blood draws were performed. Total RNA was isolated from all samples. All samples were hybridized to microarrays to examine gene expression changes over the timed exposure to ethanol. One experimental sample failed. The remaining samples for Subject 18, T1-3, T5 were included in the study. There are 29 arrays for the experimental samples and 25 arrays for the control set of samples. There are separate RMA files for the experimental series and for the control series.

ORGANISM(S): Homo sapiens

SUBMITTER: Doris Kupfer

PROVIDER: E-GEOD-20489 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Microarray characterization of gene expression changes in blood during acute ethanol exposure.

Kupfer Doris M DM White Vicky L VL Strayer David L DL Crouch Dennis J DJ Burian Dennis D

BMC medical genomics 20130725

<h4>Background</h4>As part of the civil aviation safety program to define the adverse effects of ethanol on flying performance, we performed a DNA microarray analysis of human whole blood samples from a five-time point study of subjects administered ethanol orally, followed by breathalyzer analysis, to monitor blood alcohol concentration (BAC) to discover significant gene expression changes in response to the ethanol exposure.<h4>Methods</h4>Subjects were administered either orange juice or oran ...[more]

PMID: 23883607

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Project description:Metabolites present in human blood document individual physiological states influenced by genetic, epigenetic, and lifestyle factors. Using high-resolution liquid chromatography-mass spectrometry (LC-MS), we performed nontargeted, quantitative metabolomics analysis in blood of 15 young (29 ± 4 y of age) and 15 elderly (81 ± 7 y of age) individuals. Coefficients of variation (CV = SD/mean) were obtained for 126 blood metabolites of all 30 donors. Fifty-five RBC-enriched metabolites, for which metabolomics studies have been scarce, are highlighted here. We found 14 blood compounds that show remarkable age-related increases or decreases; they include 1,5-anhydroglucitol, dimethyl-guanosine, acetyl-carnosine, carnosine, ophthalmic acid, UDP-acetyl-glucosamine, N-acetyl-arginine, N6-acetyl-lysine, pantothenate, citrulline, leucine, isoleucine, NAD+, and NADP+. Six of them are RBC-enriched, suggesting that RBC metabolomics is highly valuable for human aging research. Age differences are partly explained by a decrease in antioxidant production or increasing inefficiency of urea metabolism among the elderly. Pearson’s coefficients demonstrated that some age-related compounds are correlated, suggesting that aging affects them concomitantly. Although our CV values are mostly consistent with those CVs previously published, we here report previously unidentified CVs of 51 blood compounds. Compounds having moderate to high CV values (0.4–2.5) are often modified. Compounds having low CV values, such as ATP and glutathione, may be related to various diseases because their concentrations are strictly controlled, and changes in them would compromise health. Thus, human blood is a rich source of information about individual metabolic differences. </p> Validation of experimental procedures was performed as follows. First, we evaluated the contribution of sample handling to within-sample variation. The same blood sample preparation was injected 3x into the LC-MS at 80-min intervals. We thus obtained within-sample CVs (designated as CVwi), which were less than 0.1 in 107 of 126 compounds (85%). Only 10 compounds showed CVwi of 0.1-0.2, while 9 had CVwi >0.2. Second, we also examined sample-to-sample variation caused by sample preparation. Three samples were independently prepared from the same blood sample (one person), and CVs thus determined were designated as CVss. CVss values of HEPES and PIPES in the blood samples were very small (0.06~0.08 for HEPES and 0.04~0.08 for PIPES). The great majority (116/126=92%) of CVss were <0.3. </p> Blood samples drawn from four volunteers four times within 24 h are available under accession number MTBLS264. Whole blood metabolomic data from all 30 subjects are available under accession number MTBLS265. Plasma and RBC data from all 30 subjects can be found under MTBLS266 and MTBLS267, respectively.

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Microarray characterization of gene expression changes in blood during acute ethanol exposure

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Microarray characterization of gene expression changes in blood during acute ethanol exposure.

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