Sputum Matrix Metalloproteinases (MMP) mRNA and Montelukast
|Study Design:||Endpoint Classification: Safety/Efficacy Study
Intervention Model: Single Group Assignment
Masking: Open Label
Primary Purpose: Treatment
|Official Title:||The Effect of Montelukast Therapy on mRNA Profile of Matrix Metalloproteinases and Their Inhibitors in the Sputum of Patients With Asthma|
- Primary endpoint is the MMP and TIMP mRNA profile relative to a housekeeping gene [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
- The difference between treatment with montelukast for 8 weeks and placebo for mRNA for MMP and TIMP [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
- The difference between treatment with montelukast for 8 weeks and placebo for Spirometry - FEV1, FVC, FEV1/FVC ratio [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
- The difference between treatment with montelukast for 8 weeks and placebo for Induced sputum differential cell count [ Time Frame: 8 weeks ] [ Designated as safety issue: No ]
|Study Start Date:||July 2009|
|Study Completion Date:||December 2011|
|Primary Completion Date:||December 2011 (Final data collection date for primary outcome measure)|
Experimental: montelukast group
Identified patients with asthma to recieve Montelukast 10 mg (Merck Sharp & Dohme Ltd, Herts, UK) at 0800 am once daily for 8 weeks.
montelukast 10 mg once daily for 8 weeks
The following measurements will be performed at screening:
- Informed consent
- Clinical examination
- Induced sputum
The following will be performed after 8 weeks of study medication:
- Clinical examination
- Induced sputum
- Diary Card
This will be performed with a Microlab spirometer (Micro Medical Ltd, Rochester, Kent, UK). The procedure will be according to American Thoracic Society specifications(13).
Diary Card Data:
Patients will record their symptoms on a daily basis in the morning according to "cough", "breathlessness" and "wheeze" on a 4 point scale with 0=no symptoms and 3=maximal symptoms. A total symptom score will be calculated out of 12. Patients will also measure their peak expiratory flow on a daily basis in the morning and record the highest of three measurements. They will record that they have taken their study medication.
Sputum Induction & Examination:
Sputum will be obtained with hypertonic saline by the method described by Pizzichini et al(14) inhaling increasing concentrations of saline (3, 4 and 5%) each for 7 minutes, through a mouthpiece. After each period of inhalation, FEV1 will be measured for safety. Subjects will be asked to cough sputum into a sterile container. Total cell count of leukocytes will be obtained in a modified Neubauer haemocytometer. The cell viability will be determined by the trypan blue exclusion method. Four hundred non squamous cells will be counted in Wright-stained slides and the results will be expressed as a percentage and absolute number of the total non squamous count. Measurement of MMP-9, 12 TIMP-1 and TGFb will be performed in sputum supernatant.
Profile of mRNA of MMP and TIMPs:
Total RNA will be extracted from the cellular content of the induced sputum plug using a combination of Trizol extraction and Qiagen RNeasy spin columns in a similar way to previously described12. Quantitative RT-PCR, using previously developed primers and probes, will be used to determine the relative quantities of mRNA of MMPs and TIMPs as described12. We remain the only centre in the world to routinely profile the entire MMP and TIMP gene family in human samples. This gives an all encompassing view of the involvement of these enzymes and inhibitors in the disease process and also sheds light on potential new biomarkers. The possibility of expanding the gene profiling without the need for additional sputum collection also adds value to the research. This might include other proteinase families with roles in ECM breakdown or in inflammation.
Please refer to this study by its ClinicalTrials.gov identifier: NCT00947453
|University of East Anglia|
|Norwich, Norfolk, United Kingdom, NR47TJ|
|Principal Investigator:||Andrew M Wilson, MRCP (UK)||University of East Anglia|