Volume 23, Issue 12 pp. 1166-1172

Original Article

Expression analysis of extracellular microRNA in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis

Amit Kishore,

Amit Kishore

orcid.org/0000-0002-8051-1593

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Zdenka Navratilova,

Zdenka Navratilova

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Vitezslav Kolek,

Vitezslav Kolek

Department of Respiratory Medicine and Tuberculosis, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Eva Novosadova,

Eva Novosadova

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Klára Čépe,

Klára Čépe

Faculty of Science, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Roland M. du Bois,

Roland M. du Bois

Imperial College, London, UK

Search for more papers by this author

Martin Petrek,

Corresponding Author

Martin Petrek

[email protected]

orcid.org/0000-0002-4517-7825

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Correspondence: Martin Petrek, Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Hnevotinska 3, 77515 Olomouc, Czech Republic. Email: [email protected]Search for more papers by this author

Amit Kishore,

Amit Kishore

orcid.org/0000-0002-8051-1593

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Zdenka Navratilova,

Zdenka Navratilova

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Vitezslav Kolek,

Vitezslav Kolek

Department of Respiratory Medicine and Tuberculosis, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Eva Novosadova,

Eva Novosadova

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Klára Čépe,

Klára Čépe

Faculty of Science, Palacky University, Olomouc, Czech Republic

Search for more papers by this author

Roland M. du Bois,

Roland M. du Bois

Imperial College, London, UK

Search for more papers by this author

Martin Petrek,

Corresponding Author

Martin Petrek

[email protected]

orcid.org/0000-0002-4517-7825

Department of Pathological Physiology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

First published: 29 June 2018

https://doi.org/10.1111/resp.13364

Citations: 26

(Associate Editor: Darryl Knight; Senior Editor: Chris Grainge)

Share a link

Email
Wechat
Bluesky

ABSTRACT

Background and objective

MicroRNA (miRNA) are transcriptional regulators implicated in pulmonary sarcoidosis and packaged in extracellular vesicles (EV) during cellular communication. We characterized EV and investigated miRNA expression in bronchoalveolar lavage (BAL) fluid from sarcoidosis patients.

Methods

EV were characterized for size(s) using dynamic light scattering and transmission electron microscopy (TEM) analysis and protein markers by immunoblotting. Twelve extracellular and 5 cellular miRNA were investigated in BAL from 16 chest X-ray stage-I (CXR-I) and 17 CXR stage-II (CXR-II) sarcoidosis patients. Associations between miRNA and disease characteristics (extrapulmonary involvement, pulmonary function and BAL cell profile) were statistically analysed.

Results

BAL from sarcoidosis patients contained exosomes and microvesicles (MV) as EV. In these EV, expression of miR-146a (P = 0.007), miR-150 (P = 0.003) and BAL cellular miR-21 (P = 0.01) was increased in CXR-II compared with CXR-I. Other detected EV (miR-21 and miR-26a) and cellular (miR-31, miR-129-3p, miR-146a and miR-452) miRNA were not differentially expressed. The investigated miRNA did not reflect extrapulmonary involvement, but EV miR-146a and miR-150 were negatively correlated with pulmonary function (miR-146a with vital capacity (VC; Spearman's correlation coefficient (r_s), P = −0.657, 0.007), percent predicted forced expiratory volume in 1 s (FEV₁; −0.662, 0.006) and FEV₁/forced vital capacity (FVC) ratio (−0.649, 0.008); miR-150 correlated negatively with VC (−0.584, 0.019) and FEV₁/FVC ratio (−0.746, 0.001) in CXR-II cases).

Conclusion

Our data provide evidence that exosomes and microvesicles as extracellular vesicles are present in the bronchoalveolar space of sarcoidosis patients and they differentially express EV miRNA (miR-146a and miR-150), the expression of which correlates negatively with pulmonary function indices. The significance of these findings for disease pathophysiology and clinical course require further investigation.

Supporting Information

Filename

Description

resp13364-sup-0001-AppendixS1.docxWord 2007 document , 368.5 KB

Appendix S1 Additional methods’ details.

Figure S1 Spearman's correlation coefficient analysis of significantly expressed BAL miRNA with lung function indices (VC, FEV₁ (% predicted) and FEV₁/FVC) in patients with CXR-I.

Figure S2 Spearman's correlation coefficient analysis of extracellular miR-146a (left) and miR-150 (right) with lung function FEV₁/FVC ratio in sarcoidosis patients.

Table S1 Spearman correlation coefficient analysis of extracellular miR-146a and miR-150 with CD3⁺T cell numbers and cellular miR-21 with neutrophils in BAL samples from patients with CXR-I and CXR-II sarcoidosis.

Table S2 Spearman's correlation coefficient analysis for EV, cellular miRNA with the cellular profile of BAL samples and with pulmonary function indices in sarcoidosis patients.

resp13364-sup-0002-VisualAbstract.pptxPowerPoint 2007 presentation , 278.2 KB

Visual Abstract Extracellular microRNA (miRNA) expression in bronchoalveolar lavage (BAL) fluid from sarcoidosis patients.

Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

REFERENCES

1Ascoli C, Huang Y, Schott C, Turturice BA, Metwally A, Perkins DL, Finn PW. A circulating micro-RNA signature serves as a diagnostic and prognostic indicator in sarcoidosis. Am. J. Respir. Cell Mol. Biol. 2017; 58: 40–54.
10.1165/rcmb.2017-0207OC
Web of Science® Google Scholar
2Crouser ED, Julian MW, Crawford M, Shao G, Yu L, Planck SR, Rosenbaum JT, Patrick Nana-Sinkam S. Differential expression of microRNA and predicted targets in pulmonary sarcoidosis. Biochem. Biophys. Res. Commun. 2012; 417: 886–91.
10.1016/j.bbrc.2011.12.068
CAS PubMed Web of Science® Google Scholar
3Jazwa A, Kasper L, Bak M, Sobczak M, Szade K, Jozkowicz A, Sladek K, Dulak J. Differential inflammatory microRNA and cytokine expression in pulmonary sarcoidosis. Arch. Immunol. Ther. Exp. (Warsz.) 2015; 63: 139–46.
10.1007/s00005-014-0315-9
CAS PubMed Web of Science® Google Scholar
4Maertzdorf J, Weiner J, Mollenkopf H-J, TBornotTB Network, Bauer T, Prasse A, Müller-Quernheim J, Kaufmann SHE. Common patterns and disease-related signatures in tuberculosis and sarcoidosis. Proc. Natl. Acad. Sci. U. S. A. 2012; 109: 7853–8.
10.1073/pnas.1121072109
CAS PubMed Web of Science® Google Scholar
5Kiszałkiewicz J, Piotrowski WJ, Pastuszak-Lewandoska D, Górski P, Antczak A, Górski W, Domańska-Senderowska D, Migdalska-Sęk M, Czarnecka KH, Nawrot E et al. Altered miRNA expression in pulmonary sarcoidosis. BMC Med. Genet. 2016; 17: 2.
10.1186/s12881-016-0266-6
PubMed Web of Science® Google Scholar
6Petrek M, Tomankova T, Zurkova M, Kolek V, Kriegova E. MicroRNA profiling revealed downregulated miR-204 expression in bronchoalveolar cells from patients with pulmonary sarcoidosis. Am. J. Respir. Crit. Care Med. 2013; 187: A1207.
Web of Science® Google Scholar
7Dyskova T, Fillerova R, Novosad T, Kudelka M, Zurkova M, Gajdos P, Kolek V, Kriegova E. Correlation network analysis reveals relationships between microRNAs, transcription factor T-bet, and deregulated cytokine/chemokine-receptor network in pulmonary sarcoidosis. Mediators Inflamm. 2015; 2015: 121378.
10.1155/2015/121378
PubMed Web of Science® Google Scholar
8Novosadova E, Chabronova A, Kolek V, Petrek M, Navratilova Z. The serum expression of selected miRNAs in pulmonary sarcoidosis with/without Löfgren's syndrome. Mediators Inflamm. 2016; 2016: 1–12.
10.1155/2016/1246129
Web of Science® Google Scholar
9Levänen B, Bhakta NR, Torregrosa Paredes P, Barbeau R, Hiltbrunner S, Pollack JL, Sköld CM, Svartengren M, Grunewald J, Gabrielsson S et al. Altered microRNA profiles in bronchoalveolar lavage fluid exosomes in asthmatic patients. J. Allergy Clin. Immunol. 2013; 131: 894–903.
10.1016/j.jaci.2012.11.039
CAS PubMed Web of Science® Google Scholar
10Sinha A, Yadav AK, Chakraborty S, Kabra SK, Lodha R, Kumar M, Kulshreshtha A, Sethi T, Pandey R, Malik G et al. Exosome-enclosed microRNAs in exhaled breath hold potential for biomarker discovery in patients with pulmonary diseases. J. Allergy Clin. Immunol. 2013; 132: 219–22.
10.1016/j.jaci.2013.03.035
CAS PubMed Web of Science® Google Scholar
11Kulshreshtha A, Ahmad T, Agrawal A, Ghosh B. Proinflammatory role of epithelial cell-derived exosomes in allergic airway inflammation. J. Allergy Clin. Immunol. 2013; 131: 1194–203, 1203.e1–14.
10.1016/j.jaci.2012.12.1565
CAS PubMed Web of Science® Google Scholar
12Kishore A, Navratilova Z, Kolek V, Petrek M. Detection of exosomal miRNA in pulmonary sarcoidosis. Eur. Respir. J. 2014; 44: 199.
Web of Science® Google Scholar
13de Candia P, Torri A, Pagani M, Abrignani S. Serum microRNAs as biomarkers of human lymphocyte activation in health and disease. Front. Immunol. 2014; 5: 43.
10.3389/fimmu.2014.00043
PubMed Web of Science® Google Scholar
14Navratilova Z, Novosadova E, Smitalova D, Kishore A, Kolek V, Petrek M. Serum and bronchoalveolar exosomal miRNAs in pulmonary sarcoidosis. Eur. Respir. J. 2015; 46: PA3319.
Google Scholar
15Brown D, Rahman M, Nana-Sinkam SP. MicroRNAs in respiratory disease. A clinician's overview. Ann. Am. Thorac. Soc. 2014; 11: 1277–85.
10.1513/AnnalsATS.201404-179FR
PubMed Google Scholar
16Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ, Lötvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat. Cell Biol. 2007; 9: 654–9.
10.1038/ncb1596
CAS PubMed Web of Science® Google Scholar
17Mittelbrunn M, Gutiérrez-Vázquez C, Villarroya-Beltri C, González S, Sánchez-Cabo F, González MÁ, Bernad A, Sánchez-Madrid F. Unidirectional transfer of microRNA-loaded exosomes from T cells to antigen-presenting cells. Nat. Commun. 2011; 2: 282.
10.1038/ncomms1285
CAS PubMed Web of Science® Google Scholar
18Stoorvogel W. Functional transfer of microRNA by exosomes. Blood 2012; 119: 646–8.
10.1182/blood-2011-11-389478
CAS PubMed Web of Science® Google Scholar
19Qazi KR, Torregrosa Paredes P, Dahlberg B, Grunewald J, Eklund A, Gabrielsson S. Proinflammatory exosomes in bronchoalveolar lavage fluid of patients with sarcoidosis. Thorax 2010; 65: 1016–24.
10.1136/thx.2009.132027
PubMed Web of Science® Google Scholar
20Martinez-Bravo M-J, Wahlund CJE, Qazi KR, Moulder R, Lukic A, Rådmark O, Lahesmaa R, Grunewald J, Eklund A, Gabrielsson S. Pulmonary sarcoidosis is associated with exosomal vitamin D-binding protein and inflammatory molecules. J. Allergy Clin. Immunol. 2017; 139: 1186–94.
10.1016/j.jaci.2016.05.051
CAS PubMed Web of Science® Google Scholar
21Kowal J, Arras G, Colombo M, Jouve M, Morath JP, Primdal-Bengtson B, Dingli F, Loew D, Tkach M, Théry C. Proteomic comparison defines novel markers to characterize heterogeneous populations of extracellular vesicle subtypes. Proc. Natl. Acad. Sci. U. S. A. 2016; 113: E968–77.
10.1073/pnas.1521230113
CAS PubMed Web of Science® Google Scholar
22Kanada M, Bachmann MH, Hardy JW, Frimannson DO, Bronsart L, Wang A, Sylvester MD, Schmidt TL, Kaspar RL, Butte MJ et al. Differential fates of biomolecules delivered to target cells via extracellular vesicles. Proc. Natl. Acad. Sci. U. S. A. 2015; 112: E1433–42.
10.1073/pnas.1418401112
CAS PubMed Web of Science® Google Scholar
23Statement on sarcoidosis. Joint statement of the American Thoracic Society (ATS), the European Respiratory Society (ERS) and the World Association of Sarcoidosis and Other Granulomatous Disorders (WASOG) adopted by the ATS Board of Directors and by ERS Executive Committee, February 1999. Am. J. Respir. Crit. Care Med. 1999; 160: 736–55.
10.1164/ajrccm.160.2.ats4-99
CAS PubMed Web of Science® Google Scholar
24Meyer KC, Raghu G, Baughman RP, Brown KK, Costabel U, du Bois RM, Drent M, Haslam PL, Kim DS, Nagai S et al.; American Thoracic Society Committee on BAL in Interstitial Lung Disease. An official American Thoracic Society clinical practice guideline: the clinical utility of bronchoalveolar lavage cellular analysis in interstitial lung disease. Am. J. Respir. Crit. Care Med. 2012; 185: 1004–14.
Google Scholar
25Théry C, Amigorena S, Raposo G, Clayton A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol. 2006; Chapter 3: Unit 3.22.
PubMed Google Scholar
26Foster PS, Plank M, Collison A, Tay HL, Kaiko GE, Li J, Johnston SL, Hansbro PM, Kumar RK, Yang M et al. The emerging role of microRNAs in regulating immune and inflammatory responses in the lung. Immunol. Rev. 2013; 253: 198–215.
10.1111/imr.12058
CAS PubMed Web of Science® Google Scholar
27Kishore A, Borucka J, Petrkova J, Petrek M. Novel insights into miRNA in lung and heart inflammatory diseases. Mediators Inflamm. 2014; 2014: 259131.
10.1155/2014/259131
CAS PubMed Web of Science® Google Scholar
28Zhou T, Garcia JGN, Zhang W. Integrating microRNAs into a system biology approach to acute lung injury. Transl. Res. 2011; 157: 180–90.
10.1016/j.trsl.2011.01.010
CAS PubMed Web of Science® Google Scholar
29Taganov KD, Boldin MP, Chang K-J, Baltimore D. NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc. Natl. Acad. Sci. U. S. A. 2006; 103: 12481–6.
10.1073/pnas.0605298103
CAS PubMed Web of Science® Google Scholar
30Perry MM, Moschos SA, Williams AE, Shepherd NJ, Larner-Svensson HM, Lindsay MA. Rapid changes in microRNA-146a expression negatively regulate the IL-1beta-induced inflammatory response in human lung alveolar epithelial cells. J. Immunol. 2008; 180: 5689–98.
10.4049/jimmunol.180.8.5689
CAS PubMed Web of Science® Google Scholar
31Karrich JJ, Jachimowski LCM, Libouban M, Iyer A, Brandwijk K, Taanman-Kueter EW, Nagasawa M, de JEC, Uittenbogaart CH, Blom B. MicroRNA-146a regulates survival and maturation of human plasmacytoid dendritic cells. Blood 2013; 122: 3001–9.
10.1182/blood-2012-12-475087
CAS PubMed Web of Science® Google Scholar
32Ghisi M, Corradin A, Basso K, Frasson C, Serafin V, Mukherjee S, Mussolin L, Ruggero K, Bonanno L, Guffanti A et al. Modulation of microRNA expression in human T-cell development: targeting of NOTCH3 by miR-150. Blood 2011; 117: 7053–62.
10.1182/blood-2010-12-326629
CAS PubMed Web of Science® Google Scholar
33Oglesby IK, McElvaney NG, Greene CM. MicroRNAs in inflammatory lung disease – master regulators or target practice? Respir. Res. 2010; 11: 148.
10.1186/1465-9921-11-148
CAS PubMed Web of Science® Google Scholar
34Lu TX, Munitz A, Rothenberg ME. MicroRNA-21 is up-regulated in allergic airway inflammation and regulates IL-12p35 expression. J. Immunol. 2009; 182: 4994–5002.
10.4049/jimmunol.0803560
CAS PubMed Web of Science® Google Scholar
35Wu X-B, Wang M-Y, Zhu H-Y, Tang S-Q, You Y-D, Xie Y-Q. Overexpression of microRNA-21 and microRNA-126 in the patients of bronchial asthma. Int. J. Clin. Exp. Med. 2014; 7: 1307–12.
CAS PubMed Web of Science® Google Scholar
36Moschos SA, Williams AE, Perry MM, Birrell MA, Belvisi MG, Lindsay MA. Expression profiling in vivo demonstrates rapid changes in lung microRNA levels following lipopolysaccharide-induced inflammation but not in the anti-inflammatory action of glucocorticoids. BMC Genomics 2007; 8: 240.
10.1186/1471-2164-8-240
CAS PubMed Web of Science® Google Scholar
37Liu G, Friggeri A, Yang Y, Milosevic J, Ding Q, Thannickal VJ, Kaminski N, Abraham E. miR-21 mediates fibrogenic activation of pulmonary fibroblasts and lung fibrosis. J. Exp. Med. 2010; 207: 1589–97.
10.1084/jem.20100035
CAS PubMed Web of Science® Google Scholar

Citing Literature

Volume23, Issue12

December 2018

Pages 1166-1172

Expression analysis of extracellular microRNA in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis

ABSTRACT

Background and objective

Methods

Results

Conclusion

Supporting Information

REFERENCES

Citing Literature

References

Information

About Wiley Online Library

Help & Support

Opportunities

Connect with Wiley

Expression analysis of extracellular microRNA in bronchoalveolar lavage fluid from patients with pulmonary sarcoidosis

ABSTRACT

Background and objective

Methods

Results

Conclusion

Supporting Information

REFERENCES

Citing Literature

References

Related

Information