Epigenetic Regulation of Exercise Induced Asthma

Epigenetic Regulation of Excercise Induced Asthma

The purpose of this study is to investigate the micro ribonucleic acid (mRNA) profiles of patients with EIA without allergic sensitization and EIA with house dust mite sensitization compared to that of healthy controls.

Study Overview

• Status: Recruiting
• Conditions: House Dust Mite Allergy; Exercise Induced Asthma
• Intervention / Treatment: Diagnostic test: Exercise challenge in the cold chamber (ECC); Diagnostic test: mRNA profile; Diagnostic test: standard diagnostic

Detailed Description

Patients with known EIA are periodically reexamined by medical history, clinical examination, bodyplethysmography, spirometry, exhaled nitric oxide (eNO), skin prick test, methacholin challenge test and exercise-challenge in a cold-chamber at 2-4°C (ECC). Additionally the investigators gathers the questionnaires: Asthma Control Test (ACT) and Dyspnoe Index (DI).

Besides these standard procedures the investigators want to investigate the micro-RNA profiles in two EIA subgroups: EIA with house-dust allergy (n = 24) and EIA without house-dust allergy (n = 24).

Both groups are characterized by different eNO levels. The patients with EIA and house-dust allergy should have an eNO > 30 ppb, the patients with EIA without house-dust allergy an eNO < 20 ppb.

The micro-RNA profiles of the both EIA subgroups will be compared the micro RNA profiles of 20 healthy controls .

Therefore blood will be taken (for a complete blood count and micro RNA analysis) at three points of time: before ECC, directly after ECC and after 24 hours ± 4 hours after ECC.

• Study Type: Interventional
• Enrollment (Anticipated): 68
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Melanie Dreßler, MD; Phone Number: +49-69-6301-4588; Email: melanie.dressler@kgu.de
• Study Contact Backup: Name: Helena Donath, MD; Phone Number: +49-69-6301-4588; Email: helena.donath@kgu.de

Study Locations

• Germany
  • Hessen
    • Frankfurt, Hessen, Germany, 60590
      • Recruiting
      • Universitätsklinikum Frankfurt
      • Contact: Melanie Dreßler, MD; Phone Number: +49-69-6301-4588; Email: melanie.dressler@kgu.de
      • Contact: Helena Donath, MD; Phone Number: +49-69-6301-4588; Email: helena.donath@kgu.de

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 12 years to 24 years (ADULT, CHILD)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

For EIA patients

Inclusion Criteria:

• written agreement
• age >=12 and <= 24
• known exercise induced asthma
• Group 1: skin prick test positiv house-dust allergy, eNO > 30 ppb, MCT PD20 < 0,1 mg
• Group 2: skin prick test negative house-dust allergy, eNO < 20 ppb, MCT PD20 < 1 mg
• lung function before ECC forced vital capacity (FVC) ≥ 75% and forced exspiratory pressure in one second (FEV1) ≥ 70%

Exclusion Criteria:

• age < 12 und > 24 years
• lung function FVC < 75% und FEV1< 70%
• inability to understand the range of the study
• chronic asthma with systemic cortisone therapy
• regular therapy with inhalative corticosteroids or leukotriene-antagonists <14 days before visit 1
• intake of long acting beta-agonists (LABA) 48 h before examination
• intake of short acting beta-agonists (SABA) 8 h before examination
• acute severe infection (pneumonia) within the last 4 weeks
• other chronic diseases or infections (HIV, Tbc)
• pregnancy

For healthy controls

Inclusion Criteria:

• written agreement
• age >=18 and <= 24

Exclusion Criteria:

• age < 18 and > 24 years
• known asthma bronchiale or other chronic lung diseases
• lung function FVC < 90% and FEV1 < 80%
• allergic sensitization in skin prick test
• eNO > 30 ppb
• inability to understand the range of the study
• acute severe infection (pneumonia) within the last 4 weeks
• other chronic diseases or infections (HIV, Tbc)
• pregnancy

Study Plan

How is the study designed?

Design Details

• Primary Purpose: DIAGNOSTIC
• Allocation: NON_RANDOMIZED
• Interventional Model: PARALLEL
• Masking: NONE

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
OTHER: EIA wih house dust mite; Patients with EIA and house-dust mite allergy and an eNO > 30 ppb	Diagnostic test: Exercise challenge in the cold chamber (ECC); Exercise challenge (defined as running on a treadmill for 6-8 minutes on submaximal work load in a cold chamber. Gradient: 10%, temperature 2-4°C. After the exercise challenge controlling bodyplethysmography and spirometry after 5, 10, 15 and 30 minutes. A positive reaction is a decrease of ≥ 10% of FEV1 in comparison to the initial value.; Diagnostic test: mRNA profile; Blood taking with full blood exam and PAXgene tube for investigation oft the micro RNA profiles. The PAXgene Blood RNA kit insulates total RNA inclusive miRNA and small RNA which are centrifugated, pelletised, washed and incubated to detach proteins. For mapping the particular miRNA there will be generated libraries with specifical codes to match the analysis to the original probe. Next generation sequencing will be performed by MISeq REagent Kit v3 and the analysis oft the different expressions in the R Version 3.2.3. The divergent expressions will be refurbished by computer and databased.; Diagnostic test: standard diagnostic; bodyplethysmography, spirometry, exhaled NO, skin prick test, asthma control test (ACT)
OTHER: EIA without sensitization; Patients with EIA without allergic sensitization and an eNO < 20 ppb	Diagnostic test: Exercise challenge in the cold chamber (ECC); Exercise challenge (defined as running on a treadmill for 6-8 minutes on submaximal work load in a cold chamber. Gradient: 10%, temperature 2-4°C. After the exercise challenge controlling bodyplethysmography and spirometry after 5, 10, 15 and 30 minutes. A positive reaction is a decrease of ≥ 10% of FEV1 in comparison to the initial value.; Diagnostic test: mRNA profile; Blood taking with full blood exam and PAXgene tube for investigation oft the micro RNA profiles. The PAXgene Blood RNA kit insulates total RNA inclusive miRNA and small RNA which are centrifugated, pelletised, washed and incubated to detach proteins. For mapping the particular miRNA there will be generated libraries with specifical codes to match the analysis to the original probe. Next generation sequencing will be performed by MISeq REagent Kit v3 and the analysis oft the different expressions in the R Version 3.2.3. The divergent expressions will be refurbished by computer and databased.; Diagnostic test: standard diagnostic; bodyplethysmography, spirometry, exhaled NO, skin prick test, asthma control test (ACT)
OTHER: Healthy controls; Healthy controls without allergic sensitization or known asthma	Diagnostic test: mRNA profile; Blood taking with full blood exam and PAXgene tube for investigation oft the micro RNA profiles. The PAXgene Blood RNA kit insulates total RNA inclusive miRNA and small RNA which are centrifugated, pelletised, washed and incubated to detach proteins. For mapping the particular miRNA there will be generated libraries with specifical codes to match the analysis to the original probe. Next generation sequencing will be performed by MISeq REagent Kit v3 and the analysis oft the different expressions in the R Version 3.2.3. The divergent expressions will be refurbished by computer and databased.; Diagnostic test: standard diagnostic; bodyplethysmography, spirometry, exhaled NO, skin prick test, asthma control test (ACT)

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Dysregulation of miRNA-146	The primary endpoint is the dysregulation of miRNA-146 in comparison to the control group and within the subgroups which play an determined role in the bronchial inflammation. Therefore the total RNA, including miRNA, will be isolated using the PAXgene Blood miRNA Kit (Qiagen, Hilden, Germany). Concentration of RNA will be assessed using Nanodrop Lite spectrometry (Thermo Scientific, Dreieich, Germany). MiRNA libraries are generated with the QIAseq miRNA Library Kit (Qiagen, Hilden, Germany). Next generation sequencing (NGS) will be performed with the MiSeq Reagent Kit v3, the PhiX Sequencing Control v3 and the MiSeq™ Desktop Sequencer (all Illumina Inc., San Diego, CA, USA). Differential expression analysis will be performed in RStudio 1.2.1335 (https://cran.r-project.org/). False discovery rate correction will be applied and miRNAs are considered to be differentially expressed with p < 0.05.	1 Year

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Dysregulation miRNAs affecting Th2 mediated bronchial inflammation (miRNA-126, miRNA-21, miRNA-145, let7-mimic)	Dysregulation of other miRNAs affecting the Th2 mediated bronchial inflammation (miRNA-126, miRNA-21, miRNA-145, let7-mimic) and compare to the control group. MiRNA preparation and measurement processes are described in Outcome 1.	1 Year
Cange of eNO	Change of eNO after ECC in the EIA subgroups compared to the healthy controls	1 Year
Change of leucocytes and neutrophile granulocytes	Change of leucocytes and neutrophile granulocytes after ECC in the EIA subgroups compared to the healthy controls	1 Year
FEV1-decrease in ECC	Comparison of the FEV1-decrease in ECC with significant dysregulated miRNA in the EIA subgroups compared to the healthy controls	1 Year
Comparison eNO with significant dysregulated miRNA	Comparison of the initial value of exhaled NO with significant dysregulated miRNA in the EIA subgroups compared to the healthy controls	1 Year
Comparison of change of leucocytes and neutrophile granulocytes with significant dysregulated miRNA	Comparison of the increase of leucocytes and neutrophile granulocytes after ECC significant dysregulated miRNA in the EIA subgroups compared to the healthy controls	1 Year
Comparison of ACT with significant dysregulated miRNA	Comparison of the score in ACT with significant dysregulated miRNA in the EIA subgroups compared to the healthy controls	1 Year
Comparison of DI with significant dysregulated miRNA	Comparison of the scoresin DI with significant dysregulated miRNA in the EIA subgroups compared to the healthy controls	1 Year

Collaborators and Investigators

• Sponsor: Johann Wolfgang Goethe University Hospital
• Investigators: Principal Investigator: Melanie Dreßler, MD, Johann Wolfgang Goethe University Hospital

Publications and helpful links

General Publications

• Parsons JP, Hallstrand TS, Mastronarde JG, Kaminsky DA, Rundell KW, Hull JH, Storms WW, Weiler JM, Cheek FM, Wilson KC, Anderson SD; American Thoracic Society Subcommittee on Exercise-induced Bronchoconstriction. An official American Thoracic Society clinical practice guideline: exercise-induced bronchoconstriction. Am J Respir Crit Care Med. 2013 May 1;187(9):1016-27. doi: 10.1164/rccm.201303-0437ST.
• Dressler M, Friedrich T, Lasowski N, Herrmann E, Zielen S, Schulze J. Predictors and reproducibility of exercise-induced bronchoconstriction in cold air. BMC Pulm Med. 2019 May 16;19(1):94. doi: 10.1186/s12890-019-0845-3.
• Dressler M, Salzmann-Manrique E, Zielen S, Schulze J. Exhaled NO as a predictor of exercise-induced asthma in cold air. Nitric Oxide. 2018 Jun 1;76:45-52. doi: 10.1016/j.niox.2018.03.004. Epub 2018 Mar 8.
• Schulze J, Smith HJ, Fuchs J, Herrmann E, Dressler M, Rose MA, Zielen S. Methacholine challenge in young children as evaluated by spirometry and impulse oscillometry. Respir Med. 2012 May;106(5):627-34. doi: 10.1016/j.rmed.2012.01.007. Epub 2012 Feb 10.
• Schulze J, Rosewich M, Riemer C, Dressler M, Rose MA, Zielen S. Methacholine challenge--comparison of an ATS protocol to a new rapid single concentration technique. Respir Med. 2009 Dec;103(12):1898-903. doi: 10.1016/j.rmed.2009.06.007. Epub 2009 Jul 10.
• Schulze J, Rosewich M, Dressler M, Riemer C, Rose MA, Zielen S. Bronchial allergen challenge using the Medicaid dosimeter. Int Arch Allergy Immunol. 2012;157(1):89-97. doi: 10.1159/000324473. Epub 2011 Sep 7.
• Schulze J, Voss S, Zissler U, Rose MA, Zielen S, Schubert R. Airway responses and inflammation in subjects with asthma after four days of repeated high-single-dose allergen challenge. Respir Res. 2012 Sep 19;13(1):78. doi: 10.1186/1465-9921-13-78.
• Anderson SD, Charlton B, Weiler JM, Nichols S, Spector SL, Pearlman DS; A305 Study Group. Comparison of mannitol and methacholine to predict exercise-induced bronchoconstriction and a clinical diagnosis of asthma. Respir Res. 2009 Jan 23;10(1):4. doi: 10.1186/1465-9921-10-4.
• Driessen JM, van der Palen J, van Aalderen WM, de Jongh FH, Thio BJ. Inspiratory airflow limitation after exercise challenge in cold air in asthmatic children. Respir Med. 2012 Oct;106(10):1362-8. doi: 10.1016/j.rmed.2012.06.017. Epub 2012 Jul 11.
• Zielen S, Lieb A, De La Motte S, Wagner F, de Monchy J, Fuhr R, Munzu C, Koehne-Voss S, Riviere GJ, Kaiser G, Erpenbeck VJ. Omalizumab protects against allergen- induced bronchoconstriction in allergic (immunoglobulin E-mediated) asthma. Int Arch Allergy Immunol. 2013;160(1):102-10. doi: 10.1159/000339243. Epub 2012 Aug 28.
• Rosewich M, Zissler UM, Kheiri T, Voss S, Eickmeier O, Schulze J, Herrmann E, Ducker RP, Schubert R, Zielen S. Airway inflammation in children and adolescents with bronchiolitis obliterans. Cytokine. 2015 May;73(1):156-62. doi: 10.1016/j.cyto.2014.10.026. Epub 2015 Mar 6.
• Jerkic SP, Michel F, Donath H, Herrmann E, Schubert R, Rosewich M, Zielen S. Calprotectin as a New Sensitive Marker of Neutrophilic Inflammation in Patients with Bronchiolitis Obliterans. Mediators Inflamm. 2020 May 1;2020:4641585. doi: 10.1155/2020/4641585. eCollection 2020.
• Fussbroich D, Kohnle C, Schwenger T, Driessler C, Ducker RP, Eickmeier O, Gottwald G, Jerkic SP, Zielen S, Kreyenberg H, Beermann C, Chiocchetti AG, Schubert R. A combination of LCPUFAs regulates the expression of miRNA-146a-5p in a murine asthma model and human alveolar cells. Prostaglandins Other Lipid Mediat. 2020 Apr;147:106378. doi: 10.1016/j.prostaglandins.2019.106378. Epub 2019 Nov 4.
• Tan Z, Randall G, Fan J, Camoretti-Mercado B, Brockman-Schneider R, Pan L, Solway J, Gern JE, Lemanske RF, Nicolae D, Ober C. Allele-specific targeting of microRNAs to HLA-G and risk of asthma. Am J Hum Genet. 2007 Oct;81(4):829-34. doi: 10.1086/521200. Epub 2007 Aug 20. Erratum In: Am J Hum Genet. 2008 Jan;82(1):251.
• Zhang YY, Zhong M, Zhang MY, Lv K. [Expression and clinical significance of miR-155 in peripheral blood CD4(+);T cells of patients with allergic asthma]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2012 May;28(5):540-3. Chinese.
• Lu TX, Munitz A, Rothenberg ME. MicroRNA-21 is up-regulated in allergic airway inflammation and regulates IL-12p35 expression. J Immunol. 2009 Apr 15;182(8):4994-5002. doi: 10.4049/jimmunol.0803560.
• Mattes J, Collison A, Plank M, Phipps S, Foster PS. Antagonism of microRNA-126 suppresses the effector function of TH2 cells and the development of allergic airways disease. Proc Natl Acad Sci U S A. 2009 Nov 3;106(44):18704-9. doi: 10.1073/pnas.0905063106. Epub 2009 Oct 20.
• Williams AE, Larner-Svensson H, Perry MM, Campbell GA, Herrick SE, Adcock IM, Erjefalt JS, Chung KF, Lindsay MA. MicroRNA expression profiling in mild asthmatic human airways and effect of corticosteroid therapy. PLoS One. 2009 Jun 12;4(6):e5889. doi: 10.1371/journal.pone.0005889.
• Polikepahad S, Knight JM, Naghavi AO, Oplt T, Creighton CJ, Shaw C, Benham AL, Kim J, Soibam B, Harris RA, Coarfa C, Zariff A, Milosavljevic A, Batts LM, Kheradmand F, Gunaratne PH, Corry DB. Proinflammatory role for let-7 microRNAS in experimental asthma. J Biol Chem. 2010 Sep 24;285(39):30139-49. doi: 10.1074/jbc.M110.145698. Epub 2010 Jul 14.
• Chiba Y, Tanabe M, Goto K, Sakai H, Misawa M. Down-regulation of miR-133a contributes to up-regulation of Rhoa in bronchial smooth muscle cells. Am J Respir Crit Care Med. 2009 Oct 15;180(8):713-9. doi: 10.1164/rccm.200903-0325OC. Epub 2009 Jul 30.
• Kumar M, Mabalirajan U, Agrawal A, Ghosh B. Proinflammatory role of let-7 miRNAs in experimental asthma? J Biol Chem. 2010 Nov 26;285(48):le19: author reply le20. doi: 10.1074/jbc.L110.145698. No abstract available.
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• Liu X, Nelson A, Wang X, Kanaji N, Kim M, Sato T, Nakanishi M, Li Y, Sun J, Michalski J, Patil A, Basma H, Rennard SI. MicroRNA-146a modulates human bronchial epithelial cell survival in response to the cytokine-induced apoptosis. Biochem Biophys Res Commun. 2009 Feb 27;380(1):177-82. doi: 10.1016/j.bbrc.2009.01.066. Epub 2009 Jan 22.
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Study record dates

Study Major Dates

• Study Start (ACTUAL): August 1, 2021
• Primary Completion (ANTICIPATED): August 1, 2022
• Study Completion (ANTICIPATED): September 1, 2022

Study Registration Dates

• First Submitted: August 30, 2021
• First Submitted That Met QC Criteria: December 14, 2021
• First Posted (ACTUAL): January 3, 2022

Study Record Updates

• Last Update Posted (ACTUAL): January 3, 2022
• Last Update Submitted That Met QC Criteria: December 14, 2021
• Last Verified: December 1, 2021

More Information

Terms related to this study

• Keywords: eNO; Exercise Induced Asthma; exercise challenge; micro RNAs
• Additional Relevant MeSH Terms: Respiratory Tract Diseases; Immune System Diseases; Lung Diseases; Hypersensitivity, Immediate; Bronchial Diseases; Lung Diseases, Obstructive; Respiratory Hypersensitivity; Hypersensitivity; Asthma; Asthma, Exercise-Induced
• Other Study ID Numbers: E68/21

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No