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Airway remodeling and endobrochial gene expression in asthma
J ALLERGY CLIN IMMUNOL VOLUME 111, NUMBER 2
04
Airway Remodeling and Endobronchial Gene Expression in Asthma
P. Beckett, P. H. Howarth; RCMB Division, University of Southampton, Southampton, UNITED KINGDOM. RATIONALE: Asthma is associated with airway remodelling typified by myofibroblast transformation, increased airway collagen accumulation and epithelial activation associated with enhanced growth fcator expression.To investigate changes in gene expression within the airways underlying airway remodelling in asthma, we have compared gene expression in endobronchial biopsies from asthmatics and healthy controls. METHODS: Endobronchial biopsies were obtained at fiberoptic bronchoscopy from 11 healthy non-asthmatic subjects and 45 mild asthmatics, recieving as required short-acting beta-2-agonists as their sole therapy. The biopsies were snap frozen until mRNA extraction and reverse transcription were undertaken. Subsequently real-time RT-PCR (Taqman) was undertaken to measure the level of gene expression relative to the housekeeping gene 18s rRNA. RESULTS: There was significantly greater mRNA expression for smooth muscle-actin (p<0.04), collagen Ill (p<0.001), MMP-3 (p<0.005) and TIMP-! (p<0.001) in the asthmatic samples but there was no difference in the gene expression for bFGF, TGF-beta, endothelin, collagen 1, MMP-9 or PAl- 1. No differences were evident in the MMP-3/TIMP- 1 ratios. C O N C L U S I O N S : This study shows evidence of upregulated gene expression in relationship to fibroblast activation and transformation within the airways in asthma and enhanced but balanced activation of MMP3/TIMP-1. The lack of upregulated gene expression for growth factors suggests that post-translational modification of activity rather than de novo synthesis may be more relevant to their on-going biological activity within the airways.
Funding: GlaxoSmithKline
05
Airway Responsivenessto Beta-Agonist and Beta2-Adrenoceptnr Gene Polymorphisms
K. Asano I, Y. Suzuki I, T. Shiomi I, T. Nakajima j, H. Kudoh I, T. Matsuzaki I, W. Yamada 1, R, Hiraoka I, K. Yamaguchi I, A. Nagabukuro 2, Y. Harada2; lKeio University School of Medicine, Tokyo, JAPAN, 2Otsuka Pharmaceutical Co Ltd TRC, Tokushima, JAPAN. RATIONALE: Since genetic variability has been shown to modulate the expression and/or functions of 132-adrenoceptor, we examined the association of bronchodilatory response to I~-agonist, procaterol, with [32adrenoeeptor genotype/haplotype in patients with asthma. METHODS: Ninety-eight Japanese patients with moderate to severe asthma were recruited. Increase in FEVI was measured after inhalation of procaterol (AFEVI[3) and after inhalation of procaterol + oxitropium (AFEVITolal) to determine the ~2-adrenoceptor-dependent and -independent airway responsiveness. The entire coding region (1239 bp) and 5"flanking region (-1.8 kb) of ~2-adrenoceptor gene was amplified by PCR, and directly sequenced for genotyping, or subcloned into plasmids and sequenced for haplotyping. RESULTS: Fourteen single nucleotide polymorphisms have been identified; 3 major haplotypes explained more than 80% of the alleles. None of the genotypes or haplotypes of t32-adrenoceptor was significantly associated with AFEV I~ adjusted with sex, age, and height, or with the ratio of AFEVlf~ to AFEVI.rot~ I. There was a strong correlation between AFEVI~ and AFEVITota 1 (r = 0.95, p < 0.00l), suggesting that the variability of bronchodilatory response to procaterol is mostly determined by 132adrenoceptor-independent factors. CONCLUSIONS: In Japanese patients with asthma, airway responsiveness to J3-agonist is not determined by 132-adrenoceptor gene polymorphisms, but is determined by the 132-adrenoceptor-independent factors such as smooth muscle contractility and airway remodeling.
Funding: Self-funded
Abstracts
$269
06 vsComparative Efficacyof AsthmaTreatments:Impactof Allergic Non-allergic Status on Responseto Therapy K. Riekard, P. Dorinsky, S. Yancey, A. Emmett; GlaxoSmithKline, Research Triangle Park, NC. RATIONALE: The purpose of this retrospective analysis was to evaluate whether allergic vs. non-allergic asthma patients differed in their response to asthma therapies. METHODS: A retrospective analysis of three recently published, randomized, double-blind, 12-week trials was conducted. Responses to treatment among asthma patients with allergies were compared with those of non-allergic asthma patients, Patients from two trials received fluticasone propionate (FP)/salmeterol combination (FSC) 100/50 BID (n=427) or montelukast I0 mg QD (n=428), Patients in the other trial received FSC 100/50 BID (n=222) or FP 100 meg + montelukast 10 mg QD (n=225). RESULTS: There were no differences in treatment responses in allergic vs. non-allergic patients for any treatment group in any study. Overall improvements from baseline in AM PEF were 82.7 vs. 88.5 L/min in allergic and non-allergic patients treated with FSC compared with 39.8 vs. 35.7 L/min in allergic and non-allergic patients treated with montelukast, Likewise, improvements from baseline in AM PEF were 29.1 vs. 24.8 L/min in allergic and non-allergic patients treated with FSC compared with 11.1 vs. 12.0 L/rain in allergic and non-allergic patients treated with FP 100 mcg + montelukast. Similar results were seen for other endpoints including FEVt, symptom-free days and albuterot use. C O N C L U S I O N S : There was no evidence that FSC or montelukast, whether given as an initial maintenance treatment or as add-on therapy to FP, was associated with differential effectiveness in allergic vs. non-allergic patients with asthma. In addition, improvements in overall asthma control were greater following treatment with FSC than with montelukast or FP 100 mcg + montelukast.
Funding: GlaxoSmithKline
807 Lower-Leg Growth in Children wi,h Asthma DuringTreatment with Inhaled Corticosteroids S. Pedersen ], L. Agertoft z, T. Lee 2, H. Staudinger2; ~Department of Pediatrics, Kolding Hospital, University of Odense, Kolding, DENMARK, 2Schering-Plough Research Institute, Kenilworth, NJ. RATIONALE: Knemometry is a highly sensitive measurement of shortterm linear growth in children, although not a predictor of final adult height. A 4-way, active- and placebo-controlled, evaluator-blind crossover study was designed to determine the effects of treatment with inhaled corticosteroids (ICS) in children with mild asthma who previously used only short-acting 1~2-agonists. METHODS: Children (N=51) 6 to 12 years of age received mometasone furoate administered by dry powder inhaler (MF-DPI) 100 mcg QD pm, MF-DPI 100 mcg BID, budesonide (BUD-DPI) 200 mcg BID, or placebo. Assessment of lower-leg growth (knemometry) and efficacy were examined during 4 sequential 2-week treatment periods with intervening 2-week washout periods. RESULTS: All 3 active treatment groups showed statistically significant decreases in lower-leg growth compared with placebo during the 2-week treatment periods (P<.05). The slowest mean growth occurred in the BUD-DP1200 mcg BID group (0.544 mm _+0.598 mm) compared with MF-DPI 100 mcg BID (0.667 mm ___0.427 ram), MF-DPI 100 meg QD pm group (0.700 mm -_+ 0.472 mm), and placebo (0.886 mm _+0.490 ram). Sm',dl improvements in pulmonary function were noted with all active treatments (NS). A slight decrease relative to baseline was observed with placebo. All treatments were well tolerated, with no treatment-related adverse events reported. CONCLUSIONS: All active treatments were associated with significantly less lower-leg growth compared with placebo, with BUD-DPI treatment generating the largest difference l~'om placebo. These findings are consistent with those observed in knemometry studies evaluating other ICS. Clinical significance of this finding requires further study.
Funding: Schering Corporation
04
Airway Remodeling and Endobronchial Gene Expression in Asthma
P. Beckett, P. H. Howarth; RCMB Division, University of Southampton, Southampton, UNITED KINGDOM. RATIONALE: Asthma is associated with airway remodelling typified by myofibroblast transformation, increased airway collagen accumulation and epithelial activation associated with enhanced growth fcator expression.To investigate changes in gene expression within the airways underlying airway remodelling in asthma, we have compared gene expression in endobronchial biopsies from asthmatics and healthy controls. METHODS: Endobronchial biopsies were obtained at fiberoptic bronchoscopy from 11 healthy non-asthmatic subjects and 45 mild asthmatics, recieving as required short-acting beta-2-agonists as their sole therapy. The biopsies were snap frozen until mRNA extraction and reverse transcription were undertaken. Subsequently real-time RT-PCR (Taqman) was undertaken to measure the level of gene expression relative to the housekeeping gene 18s rRNA. RESULTS: There was significantly greater mRNA expression for smooth muscle-actin (p<0.04), collagen Ill (p<0.001), MMP-3 (p<0.005) and TIMP-! (p<0.001) in the asthmatic samples but there was no difference in the gene expression for bFGF, TGF-beta, endothelin, collagen 1, MMP-9 or PAl- 1. No differences were evident in the MMP-3/TIMP- 1 ratios. C O N C L U S I O N S : This study shows evidence of upregulated gene expression in relationship to fibroblast activation and transformation within the airways in asthma and enhanced but balanced activation of MMP3/TIMP-1. The lack of upregulated gene expression for growth factors suggests that post-translational modification of activity rather than de novo synthesis may be more relevant to their on-going biological activity within the airways.
Funding: GlaxoSmithKline
05
Airway Responsivenessto Beta-Agonist and Beta2-Adrenoceptnr Gene Polymorphisms
K. Asano I, Y. Suzuki I, T. Shiomi I, T. Nakajima j, H. Kudoh I, T. Matsuzaki I, W. Yamada 1, R, Hiraoka I, K. Yamaguchi I, A. Nagabukuro 2, Y. Harada2; lKeio University School of Medicine, Tokyo, JAPAN, 2Otsuka Pharmaceutical Co Ltd TRC, Tokushima, JAPAN. RATIONALE: Since genetic variability has been shown to modulate the expression and/or functions of 132-adrenoceptor, we examined the association of bronchodilatory response to I~-agonist, procaterol, with [32adrenoeeptor genotype/haplotype in patients with asthma. METHODS: Ninety-eight Japanese patients with moderate to severe asthma were recruited. Increase in FEVI was measured after inhalation of procaterol (AFEVI[3) and after inhalation of procaterol + oxitropium (AFEVITolal) to determine the ~2-adrenoceptor-dependent and -independent airway responsiveness. The entire coding region (1239 bp) and 5"flanking region (-1.8 kb) of ~2-adrenoceptor gene was amplified by PCR, and directly sequenced for genotyping, or subcloned into plasmids and sequenced for haplotyping. RESULTS: Fourteen single nucleotide polymorphisms have been identified; 3 major haplotypes explained more than 80% of the alleles. None of the genotypes or haplotypes of t32-adrenoceptor was significantly associated with AFEV I~ adjusted with sex, age, and height, or with the ratio of AFEVlf~ to AFEVI.rot~ I. There was a strong correlation between AFEVI~ and AFEVITota 1 (r = 0.95, p < 0.00l), suggesting that the variability of bronchodilatory response to procaterol is mostly determined by 132adrenoceptor-independent factors. CONCLUSIONS: In Japanese patients with asthma, airway responsiveness to J3-agonist is not determined by 132-adrenoceptor gene polymorphisms, but is determined by the 132-adrenoceptor-independent factors such as smooth muscle contractility and airway remodeling.
Funding: Self-funded
Abstracts
$269
06 vsComparative Efficacyof AsthmaTreatments:Impactof Allergic Non-allergic Status on Responseto Therapy K. Riekard, P. Dorinsky, S. Yancey, A. Emmett; GlaxoSmithKline, Research Triangle Park, NC. RATIONALE: The purpose of this retrospective analysis was to evaluate whether allergic vs. non-allergic asthma patients differed in their response to asthma therapies. METHODS: A retrospective analysis of three recently published, randomized, double-blind, 12-week trials was conducted. Responses to treatment among asthma patients with allergies were compared with those of non-allergic asthma patients, Patients from two trials received fluticasone propionate (FP)/salmeterol combination (FSC) 100/50 BID (n=427) or montelukast I0 mg QD (n=428), Patients in the other trial received FSC 100/50 BID (n=222) or FP 100 meg + montelukast 10 mg QD (n=225). RESULTS: There were no differences in treatment responses in allergic vs. non-allergic patients for any treatment group in any study. Overall improvements from baseline in AM PEF were 82.7 vs. 88.5 L/min in allergic and non-allergic patients treated with FSC compared with 39.8 vs. 35.7 L/min in allergic and non-allergic patients treated with montelukast, Likewise, improvements from baseline in AM PEF were 29.1 vs. 24.8 L/min in allergic and non-allergic patients treated with FSC compared with 11.1 vs. 12.0 L/rain in allergic and non-allergic patients treated with FP 100 mcg + montelukast. Similar results were seen for other endpoints including FEVt, symptom-free days and albuterot use. C O N C L U S I O N S : There was no evidence that FSC or montelukast, whether given as an initial maintenance treatment or as add-on therapy to FP, was associated with differential effectiveness in allergic vs. non-allergic patients with asthma. In addition, improvements in overall asthma control were greater following treatment with FSC than with montelukast or FP 100 mcg + montelukast.
Funding: GlaxoSmithKline
807 Lower-Leg Growth in Children wi,h Asthma DuringTreatment with Inhaled Corticosteroids S. Pedersen ], L. Agertoft z, T. Lee 2, H. Staudinger2; ~Department of Pediatrics, Kolding Hospital, University of Odense, Kolding, DENMARK, 2Schering-Plough Research Institute, Kenilworth, NJ. RATIONALE: Knemometry is a highly sensitive measurement of shortterm linear growth in children, although not a predictor of final adult height. A 4-way, active- and placebo-controlled, evaluator-blind crossover study was designed to determine the effects of treatment with inhaled corticosteroids (ICS) in children with mild asthma who previously used only short-acting 1~2-agonists. METHODS: Children (N=51) 6 to 12 years of age received mometasone furoate administered by dry powder inhaler (MF-DPI) 100 mcg QD pm, MF-DPI 100 mcg BID, budesonide (BUD-DPI) 200 mcg BID, or placebo. Assessment of lower-leg growth (knemometry) and efficacy were examined during 4 sequential 2-week treatment periods with intervening 2-week washout periods. RESULTS: All 3 active treatment groups showed statistically significant decreases in lower-leg growth compared with placebo during the 2-week treatment periods (P<.05). The slowest mean growth occurred in the BUD-DP1200 mcg BID group (0.544 mm _+0.598 mm) compared with MF-DPI 100 mcg BID (0.667 mm ___0.427 ram), MF-DPI 100 meg QD pm group (0.700 mm -_+ 0.472 mm), and placebo (0.886 mm _+0.490 ram). Sm',dl improvements in pulmonary function were noted with all active treatments (NS). A slight decrease relative to baseline was observed with placebo. All treatments were well tolerated, with no treatment-related adverse events reported. CONCLUSIONS: All active treatments were associated with significantly less lower-leg growth compared with placebo, with BUD-DPI treatment generating the largest difference l~'om placebo. These findings are consistent with those observed in knemometry studies evaluating other ICS. Clinical significance of this finding requires further study.
Funding: Schering Corporation