Sputum Matrix Metalloproteinases (MMP) mRNA and Montelukast | RxWiki

Sputum Matrix Metalloproteinases (MMP) mRNA and Montelukast

Overview[ - collapse ][ - ]

Purpose	Matrix metalloproteinases (MMPs) are a group of 24 zinc containing enzymes in man. These enzymes were originally described as cleaving extracellular matrix (ECM) substrates with a predominant role in ECM homeostasis, but it is now clear that they have much wider functionality. An imbalance between MMP activity and that of their inhibitors (tissue inhibitors of metalloproteinases, TIMPs) is considered to play a critical role in the synthesis or degradation of the extracellular matrix of the airway architecture which results in fixed airflow obstruction in both asthma and chronic obstructive pulmonary disease (COPD). Using quantitative real time polymerase chain reaction (RT-PCR) the investigators have identified a difference between the level of steady state mRNA for MMP-9, MMP-14 and MMP-2 in 2 patients with asthma compared to 4 healthy controls using our method. However the investigators require further refinement of the process in order to optimise RNA quality and to evaluate the effect of montelukast across the entire family of MMPs and their inhibitors (TIMPs).
Condition	Asthma
Intervention	Drug: montelukast
Phase	Phase 2
Sponsor	University of East Anglia
Responsible Party	University of East Anglia
ClinicalTrials.gov Identifier	NCT00947453
First Received	July 24, 2009
Last Updated	August 22, 2012
Last verified	August 2012

Tracking Information[ + expand ][ + ]

First Received Date	July 24, 2009
Last Updated Date	August 22, 2012
Start Date	July 2009
Estimated Primary Completion Date	December 2011
Current Primary Outcome Measures	Primary endpoint is the MMP and TIMP mRNA profile relative to a housekeeping gene [Time Frame: 8 weeks] [Designated as safety issue: No]
Current Secondary Outcome Measures	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]

Descriptive Information[ + expand ][ + ]

Brief Title	Sputum Matrix Metalloproteinases (MMP) mRNA and Montelukast
Official Title	The Effect of Montelukast Therapy on mRNA Profile of Matrix Metalloproteinases and Their Inhibitors in the Sputum of Patients With Asthma
Brief Summary	Matrix metalloproteinases (MMPs) are a group of 24 zinc containing enzymes in man. These enzymes were originally described as cleaving extracellular matrix (ECM) substrates with a predominant role in ECM homeostasis, but it is now clear that they have much wider functionality. An imbalance between MMP activity and that of their inhibitors (tissue inhibitors of metalloproteinases, TIMPs) is considered to play a critical role in the synthesis or degradation of the extracellular matrix of the airway architecture which results in fixed airflow obstruction in both asthma and chronic obstructive pulmonary disease (COPD). Using quantitative real time polymerase chain reaction (RT-PCR) the investigators have identified a difference between the level of steady state mRNA for MMP-9, MMP-14 and MMP-2 in 2 patients with asthma compared to 4 healthy controls using our method. However the investigators require further refinement of the process in order to optimise RNA quality and to evaluate the effect of montelukast across the entire family of MMPs and their inhibitors (TIMPs).
Detailed Description	The following measurements will be performed at screening: - Informed consent - Clinical examination - Spirometry - Induced sputum The following will be performed after 8 weeks of study medication: - Clinical examination - Spirometry - Induced sputum - Diary Card Spirometry: 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.
Study Type	Interventional
Study Phase	Phase 2
Study Design	Endpoint Classification: Safety/Efficacy Study, Intervention Model: Single Group Assignment, Masking: Open Label, Primary Purpose: Treatment
Condition	Asthma
Intervention	Drug: montelukast montelukast 10 mg once daily for 8 weeks
Study Arm (s)	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.

Recruitment Information[ + expand ][ + ]

Recruitment Status	Completed
Estimated Enrollment	15
Estimated Completion Date	December 2011
Estimated Primary Completion Date	December 2011
Eligibility Criteria	Inclusion Criteria: - Male or female, aged 18 to 60 years. - Diagnosed with asthma, defined as episodic chest tightness, wheezing and dyspnoea, cough. - Non-Smoker or Ex-Smoker for at least 10 years and a smoking history of less than 5 pack years. - History of asthma symptoms for more than 10years. - Receiving as required short acting bronchodilators. - Post bronchodilator FEV1 50 to 100 % predicted - Evidence of airway calibre reversibility within the previous 12 months: reversibility to salbutamol of 12% following 400mcg inhaled salbutamol, histamine PC20 < 8mg/ml, diurnal variation in peak expiratory flow of 20%. - Able to produce sputum after induction with saline. Exclusion criteria: - Cardiac or pulmonary disease other than asthma. - Respiratory infection defined as fever, nasal/sinus congestion, fatigue, cough, antibiotic use or yellow/green sputum within 4 weeks prior to study. - Receiving inhaled or oral corticosteroid therapy, long acting Beta2 agonist therapy or leukotriene modifying therapy for the previous 1 month. - Severe or uncontrolled co-morbid disease. - Pregnancy or breastfeeding. - Unable to give written informed consent
Gender	Both
Ages	18 Years
Accepts Healthy Volunteers	No
Contacts	Not Provided
Location Countries	United Kingdom

Administrative Information[ + expand ][ + ]

NCT Number	NCT00947453
Other Study ID Numbers	2009RESP01
Has Data Monitoring Committee	No
Information Provided By	University of East Anglia
Study Sponsor	University of East Anglia
Collaborators	Clinical Research and Trials Unit (Norfolk & Norwich University Hospital, UK)
Investigators	Principal Investigator: Andrew M Wilson, MRCP (UK) University of East Anglia
Verification Date	August 2012

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