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| Status |
Public on Oct 20, 2021 |
| Title |
Expression profiles of epicardial and subcutaneous adipose tissue |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
The epicardial adipose tissue (EAT) is a visceral adipose tissue in close contact with coronary vessels whose increase is associated with coronary artery disease (CAD). Our goal was to identify candidate molecule(s) characterizing EAT which could intervene in the pathogenesis of CAD. An approach combining microarrays and bioinformatic sequence analysis tools for predicting secreted proteins (TargetP) was applied to paired biopsies of subcutaneous adipose tissue (SAT) and EAT, obtained from patients with or without CAD (NCAD). Results were validated in 3 independent groups of subjects by RT-qPCR, western blot, immuno histochemistry and explants secretion. sPLA2-IIA ranked first among genes coding for potentially secreted proteins with the highest overexpression in EAT in both CAD and NCAD. RT-qPCR confirmed its increased expression in EAT ( p<0.01) and in EAT from CAD as compared to NCAD (49.3 ±13 vs. 17.4 ± 9.7 p<0.01). sPLA2-IIA protein level was higher in EAT than in SAT. EAT explants demonstrated also significantly higher sPLA2-IIA secretion levels than SAT ones (4.37±2.7 vs 0.67± 0.28 ng.ml-1/g tissue/24h,p<0.03). sPLA2-IIA labeling was evidenced in EAT in the stroma vascular fraction between adipocytes and in connective capsules, with no immunostaining of the adipocytes. SAT was weakly labeled following the same pattern.
Conclusion: We evidenced for the first time an increased expression of sPLA2-IIA in EAT from patients with CAD, a phospholipase that was shown to be an independent risk factor for CAD. These findings suggest a potentially pathophysiological role of EAT in CAD.
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| Overall design |
We studied three independent groups of subjects. Group 1 and 2 were composed of 30 subjects who underwent cardiac surgery for coronary artery disease (CAD) or for valve replacement, or myxoma resection (NCAD). Patients from the NCAD group had no clinical or angiographic sign of CAD, and had a left ventricular ejection fraction > 50%. Group 3 was composed of 15 non diabetic patients with CAD. EAT and SAT samples were obtained within 20 min after the start of surgery respectively near the proximal tract of the right coronary artery and the site of chest incision. Samples were frozen in liquid nitrogen and stored at -80 °C. In groups 1 and 2, total RNA obtained from SAT and EAT samples of 7 subjects (4 CAD and 3 NCAD) was prepared using the RNeasy total RNA Mini kit (Qiagen). RNA concentration and integrity were assessed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Massy, France). For microarray experiments, 1 µg of total RNA from each total RNA sample preparation was amplified by MessageAmp RNA Kit (Ambion, Austin, TX, USA) and 3 µg of amplified RNA was labeled with cyanin dyes (Cy) using the CyScribe First-Strand cDNA labeling kit (Amersham Biosciences, Orsay, France). (available at http://cmgm.stanford.edu/pbrown/protocols/index). Using pangenomic Agilent® cDNA microarrays, we compared the expression profiles of EAT and SAT from CAD and NCAD subjects. To this purpose we used a common reference pool that was generated by mixing equal amounts of total RNA extracted from adipose tissue samples of patients undergoing plastic surgery. Amplified RNA (aRNA) from the reference pool was labeled with Cy3, and aRNA from the 7 subjects (either EAT or SAT) was labeled with Cy5. A total of 13 pangenomic microarray hybridizations were performed for this differential comparison. After scanning the 13 arrays, the images were analyzed and data were normalized using the Stanford Microarray Database Online resources (http://genomewww5.stanford.edu/MicroArray/SMD/).
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| Contributor(s) |
Henegar C, Clement K, Pelloux V, Dutour A, Achard V, Silaghi A, Jacques F |
| Citation(s) |
20008021 |
| Submission date |
Oct 17, 2009 |
| Last update date |
Oct 20, 2021 |
| Contact name |
Corneliu Henegar |
| E-mail(s) |
corneliu@henegar.info, flavien.jacques@upmc.fr
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| Organization name |
Institut national de la santé et de la recherche médicale
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| Department |
U872 Eq7
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| Lab |
Eq7 Nutriomique
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| Street address |
15 Rue de l'Ecole de Medecine
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| City |
Paris |
| State/province |
Ile de France |
| ZIP/Postal code |
75005 |
| Country |
France |
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| Platforms (1) |
| GPL4133 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Feature Number version) |
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| Samples (13)
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| Relations |
| BioProject |
PRJNA121441 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE18612_RAW.tar |
88.3 Mb |
(http)(custom) |
TAR (of GPR) |
| Processed data included within Sample table |
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