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Transcriptional profiling identifies the long noncoding RNA plasmacytoma variant translocation (PVT1) as a novel regulator of the asthmatic phenotype.

Published bySucianty Yuwono Modified over 6 years ago

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Presentation on theme: "Transcriptional profiling identifies the long noncoding RNA plasmacytoma variant translocation (PVT1) as a novel regulator of the asthmatic phenotype."— Presentation transcript:

1 Transcriptional profiling identifies the long noncoding RNA plasmacytoma variant translocation (PVT1) as a novel regulator of the asthmatic phenotype in human airway smooth muscle  Philip J. Austin, MSc, Eleni Tsitsiou, PhD, Charlotte Boardman, MD, Simon W. Jones, PhD, Mark A. Lindsay, PhD, Ian M. Adcock, PhD, Kian Fan Chung, MD, PhD, Mark M. Perry, PhD  Journal of Allergy and Clinical Immunology  Volume 139, Issue 3, Pages (March 2017) DOI: /j.jaci Copyright © 2016 The Authors Terms and Conditions

2 Fig 1 Venn diagrams showing pathway analysis of differentially expressed mRNAs in ASM from asthmatic patients. A-C, Pathway analysis showing intergroup comparisons at baseline (Fig 1, A), after stimulation with FCS (2.5%; Fig 1, B), and after stimulation with FCS (2.5%) plus TGF-β (1 ng/mL; Fig 1, C). Data represent 9 members of each patient type. D, Expression of 9 mRNAs was confirmed by using TaqMan RT-PCR to validate the array data. Bars represent means ± SEMs from 9 primary ASMC donors. *P < .05, **P < .01, and ***P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

3 Fig 2 Effect of dexamethasone and FCS plus TGF-β on PVT1 lncRNA expression in ASMCs from asthmatic patients. A, FCS plus TGF-β–induced PVT1 lncRNA expression was measured by using quantitative RT-PCR over 1, 3, 6, and 24 hours. B, Dexamethasone (Dex) and FCS plus TGF-β–induced PVT1 expression was measured by using quantitative RT-PCR at 24 hours. Points/bars represent means ± SEMs of 9 ASMC donors. **P < .01 and ***P < .001. ##P < .01. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

4 Fig 3 Effect of targeting PVT1 with siRNAs on IL-6 release and BrdU incorporation in ASMCs from healthy subjects and patients with severe asthma. A and B, PVT1 lncRNA expression was measured by using RT-PCR after ASM from healthy subjects and patients with severe asthma was transfected with siRNA (300 nmol/L), which was designed to target PVT1. C-F, BrdU incorporation (Fig 3, C and D) and IL-6 release (Fig 3, E and F) were measured by using the BrdU ELISA and DuoSet ELISA, respectively, at 8 days. Bars/points represent means ± SEMs of 9 ASMC donors. */#P < .05, ##/++P < .01, and ***/###P < .001. Dex, Dexamethasone. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

5 Fig 4 Effect of targeting PVT1 with siRNAs on IL6 and c-MYC mRNA in ASMCs from healthy subjects and patients with severe asthma. A and B, IL6 and c-MYC mRNA expression was measured by using RT-PCR after exposure to dexamethasone (Dex; 10−7 mol/L) and stimulation with FCS (2.5%) plus TGF-β (1 ng/mL) for 24 hours. C-F, ASMCs from healthy subjects and patients with severe asthma were transfected with siRNAs designed to target PVT1. Also, the expression of IL6 (Fig 4, C and E) and c-MYC (Fig 4, D and F) mRNA was measured by using RT-PCR. Bars represent means ± SEMs of 9 ASMC donors. #P < .05, **/##/++P < .01, and ***/###/+++P < .001. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

6 Fig 5 A and B, Potential mechanisms for PVT1 contribution to ASMC proliferation and IL-6 release in asthmatic patients. Dex, Dexamethasone; FCS, fetal calf serum; TGF-β, transforming growth factor-beta. Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

7 Fig E1 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

8 Fig E2 Journal of Allergy and Clinical Immunology  , DOI: ( /j.jaci ) Copyright © 2016 The Authors Terms and Conditions

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