Source: International Congress 2014 – Novel epigenetic regulators for targeting via nanomedicine
Disease area: Interstitial lung diseases

Abstract

Background: MicroRNAs (miRNAs) are implicated in pathogenesis of pulmonary sarcoidosis, namely in posttranscriptional regulation of (pro)inflammatory cytokines. Circulatory miRNAs are stable miRNAs packaged in exosomes during cellular communication that protect miRNA molecules from degradation. There has been no information on the presence of exosomal miRNA in sarcoid lung. We have, therefore, investigated 11candidate miRNAs in exosomes from sarcoid bronchoalveolar (BAL) space. Method: Exosomes were isolated from BAL fluid (BALF) from sarcoidosis patients (n=39) by differential centrifugation (3000g followed by 10000g) for 30 min each at 4°C. The supernatant was 0.2 µm filtered and centrifuged at 140000g (2 hrs at 4°C). From the obtained exosomal pellet, miRNAs was isolated and quantified using RT-qPCR. The expression data was normalized to spike-in control and BALF input volume.Result: In BALF exosomes, miR-21, -26a and -146a expression was detected with concentration (ng/µl)/ sample volume (ml) ratio ranging from 0.004-0.116 (Ct=16.4-27.3) for miR-21; 0.002-0.225 (18.9-26.9) miR-26a and 0.002-0.185 (17.9-25.0) miR-146a. Sub-analysis comparing non-smokers in radiological stage I (n=6) with stage II (n=17) showed no difference in their expression (p>0.05). MiR-15a, -129, -133a, -133b, -134, -195, -452 and -589 were not detectable in BALF exosomes. Conclusions: The implemented methodology for detection of miRNA in exosomes from BALF enabled to detect exosomal miR-21, -26a and -146a in sarcoidosis. Future analyses will focus on detailed characterization of exosomal expression profile in sarcoidosis, also in comparison with healthy subjects / diffuse lung disease. Grant support: CZ.1.07/2.3.00/30.0041, CZ.1.05./2.1.00/01.0030.

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Citations should be made in the following way:
A. Kishore, Z. Navratilova, V. Kolek, M. Petrek (Olomouc, Czech Republic). Detection of exosomal miRNA in pulmonary sarcoidosis. Eur Respir J 2014; 44: Suppl. 58, 199

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