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Soluble CD40 ligand in pulmonary arterial hypertension - possible pathogenic role of interaction between platelets and endothelial cells
S54
Abstracts
enhance recovery is to combine LVAD support with pharmacologic agents to maximise reverse remodelling followed by the selective 2-adreneric agonist clenbuterol (CLEN) to induce a mild hypertrophy and improve cardiac function (the ‘Harefield Regime’). We previously showed that CLEN given systemically to rats induces mild physiological hypertrophy with normal contractile function and elevated atrial natiuretic peptide (ANP) mRNA but without change in ␣ skeletal actin (␣SkA), a marker of pathological growth. This study examines the direct effects of CLEN on cardiac myocytes compared to pathological hypertrophy induced by isoproterenol (ISO) or phenyephrine (PE). Methods: Neonatal rat ventricular cardiac myocytes were cultured in serum free medium with or without CLEN (10 M) ISO (50M) or PE (10 M). Cell size was measured by image analysis, morphology by laser scanning confocal microscopy and protein incorporation by 3 H-phenylalanine incorporation. mRNA abundance was measured by real time quantitative PCR (TaqMan) using an ABI7700. Results: CLEN induced increased cell size (25% increase over control, p ⬍ 0.001), protein accumulation (30% increase, p ⬍ 0.001) and myofibrillar organisation, but growth was less than with either ISO or PE. In CLEN treated cells ANP and BNP mRNAs were elevated (1.8 and 2.1 fold p ⬍ 0.001) but there was no change in either ␣SkA or glucose transporter 1 (GLUT1). In contrast, ISO and PE increased ANP, BNP ␣SkA and GLUT1 indicating pathological growth. CLEN treated cells also showed elevated insulin like growth factor I (IGF-I) mRNA (3.34 fold, p ⬍ 0.001). Moreover, addition of either an IGF-I receptor blocking antibodies or LY49002, an inhibitor of PI3 kinase, the downstream target of the IGF-I receptor, inhibited the hypertrophic response to CLEN. Conclusion: These data show that clenbuterol acts directly on cardiac myocytes to induce a physiological form of hypertrophy and that this is mediated, at least in part, by IGF-I. 36 TRACLEER威 (BOSENTAN), FOR THE TREATMENT OF PULMONARY ARTERIAL HYPERTENSION (PAH) – 6 MONTH QUALITY OF LIFE DATA A. Keogh, P. Macdonald, T. Williams, K. McNeil, E. Gabbay, S. Proudman, P. Steele, J. Wlodarczyk, K. Trimble, St Vincent’s Hospital, Sydney, NSW, Australia; Alfred Hospital, Melbourne, VIC, Australia; Prince Charles Hospital, Brisbane, QLD, Australia; Royal Perth Hospital, Perth, WA, Australia; Royal Adelaide Hospital, Adelaide, SA, Australia; JWCS, Sydney, NSW, Australia; Actelion Pharmaceuticals Pty Ltd, Sydney, NSW, Australia Bosentan (TRACLEER威), has demonstrated efficacy in pulmonary arterial hypertension (PAH) in placebo-controlled trials and extends the time before clinical worsening. This multicentre, open-label, single-arm Australian study was designed to assess the efficacy and safety of bosentan in patients with primary PAH (PPH) and PAH due to connective tissue disease, and to measure quality of life changes (SF36 and AQOL) with treatment. Bosentan oral therapy was commenced at 62.5mg bid for 4 weeks, increasing to a 125mg bid maintenance dose. Results: An interim analysis was conducted on the results for the 82 patients, at 6 months following start of drug. PPH represented 70%, scleroderma 25% and SLE 5%. Mean age was 51 years (range 13– 81), median PAH duration was 1.4 years (mean 3.7), and 73% of patients were female. At baseline 87% had WHO Class III symptoms and 13% Class IV. By 6 months, of 82 patients starting bosentan, 7 had died, 1 was transplanted and 17 had been withdrawn. Last outcome was carried forward for the patients withdrawn and worst case score allocated for those who died. Improvements in QOL were marked and significant on SF36 (data shown) and AQOL (not shown) in both Class III and Class IV patients.
The Journal of Heart and Lung Transplantation February 2004
Conclusion: Patients treated with open label bosentan showed very significant improvements in all aspects of quality of life on SF36 and AQOL determined after 6 months of therapy. SF36
Baseline
Month 6
p Value
Physical functioning Role physical General health Vitality Social function Role emotional Bodily pain Mental health Health Transition
27.7 16.4 30.4 34.9 49.7 48.6 60.9 64.9 3.7
37.11 31.6 33.0 42.5 57.3 59.3 61.6 64.7 2.5
0.0001 0.0001 0.0468 0.0003 0.0007 0.0182 0.0126 0.01 0.0001
37 SOLUBLE CD40 LIGAND IN PULMONARY ARTERIAL HYPERTENSION - POSSIBLE PATHOGENIC ROLE OF INTERACTION BETWEEN PLATELETS AND ENDOTHELIAL CELLS J.K. Damas,1,2 K. Otterdal,1 A. Yndestad,1 H. Aass,2 S.S. Froland,1,3 S. Simonsen,2 P. Aukrust,1,3 A.K. Andreassen,2 1Research Institute for Internal Medicine, University of Oslo, Oslo, Norway; 2 Department of Cardiology, Rikshospitalet, Oslo, Norway; 3Section of Clinical Immunology and Infectious Diseases, Rikshospitalet, Oslo, Norway Background: Inflammatory processes seem to be involved in the development and progression of pulmonary arterial hypertension (PAH). CD40 ligand (L) may induce various biological responses such as matrix degradation and thrombus formation in several inflammatory disorders. We hypothesized that CD40L also could be involved in the immunopathogenic mechanisms of PAH. Methods and Results: By comparing soluble (s) CD40L levels in 32 PAH patients and 8 controls, we found several significant differences: 1) Patients with primary pulmonary hypertension (n ⫽ 13) and secondary PAH (n ⫽ 11) had increased sCD40L levels comparing controls (p ⬍ 0.05 and p ⬍ 0.01, respectively). 2) In contrast, sCD40L levels were normal in patients with chronic thromboembolic pulmonary hypertension (n ⫽ 8) and in other PAH patients using warfarin. 3) sCD40L levels were higher in arterial (femoral artery) comparing mixed venous blood (pulmonary artery), suggesting production of CD40L in the pulmonary vasculature. 4) Platelets from patients with PAH showed higher intracellular levels, enhanced spontaneous and thrombin-stimulated release of sCD40L comparing controls (p ⬍ 0.01). Finally, in vitro studies demonstrated that sCD40L induced chemokine expression (i.e. MCP-1 and IL-8) in endothelial cells. Indeed, MCP-1 and IL-8 were elevated and correlated to pulmonary vasculature resistance in PAH (p ⬍ 0.01). Surprisingly, prostacycline enhanced MCP-1 levels in PAH patients in vivo and in CD40Lstimulated HUVEC in vitro, suggesting some unfavorable effects of this medication in PAH. Conclusion: Raised platelet-derived CD40L levels in PAH patients may (i) reflect enhanced platelet activation and thrombus formation in this disorder, and (ii) induce chemokine expression in endothelial cells. These processes could contribute to the pathogenesis of PAH. 38 A PROSPECTIVE LONGITUDINAL STUDY OF AIRWAY VASCULATURE IN THE PATHOGENESIS OF BRONCHIOITIS OBLITERANS SYNDROME(BOS) S.Y. Langenbach,2 L. Zheng,2 T. McWilliams,2 B. Levvey,1 B. Orsida,2 M. Bailey,2 T.J. Williams,1, 2 G.I. Snell,1, 2 1Department of Allergy Immunology and Respiratory Medicine, Alfred Hospital, Melbourne, VIC, Australia; 2Department of Medicine, Monash University, Melbourne, VIC, Australia
Abstracts
enhance recovery is to combine LVAD support with pharmacologic agents to maximise reverse remodelling followed by the selective 2-adreneric agonist clenbuterol (CLEN) to induce a mild hypertrophy and improve cardiac function (the ‘Harefield Regime’). We previously showed that CLEN given systemically to rats induces mild physiological hypertrophy with normal contractile function and elevated atrial natiuretic peptide (ANP) mRNA but without change in ␣ skeletal actin (␣SkA), a marker of pathological growth. This study examines the direct effects of CLEN on cardiac myocytes compared to pathological hypertrophy induced by isoproterenol (ISO) or phenyephrine (PE). Methods: Neonatal rat ventricular cardiac myocytes were cultured in serum free medium with or without CLEN (10 M) ISO (50M) or PE (10 M). Cell size was measured by image analysis, morphology by laser scanning confocal microscopy and protein incorporation by 3 H-phenylalanine incorporation. mRNA abundance was measured by real time quantitative PCR (TaqMan) using an ABI7700. Results: CLEN induced increased cell size (25% increase over control, p ⬍ 0.001), protein accumulation (30% increase, p ⬍ 0.001) and myofibrillar organisation, but growth was less than with either ISO or PE. In CLEN treated cells ANP and BNP mRNAs were elevated (1.8 and 2.1 fold p ⬍ 0.001) but there was no change in either ␣SkA or glucose transporter 1 (GLUT1). In contrast, ISO and PE increased ANP, BNP ␣SkA and GLUT1 indicating pathological growth. CLEN treated cells also showed elevated insulin like growth factor I (IGF-I) mRNA (3.34 fold, p ⬍ 0.001). Moreover, addition of either an IGF-I receptor blocking antibodies or LY49002, an inhibitor of PI3 kinase, the downstream target of the IGF-I receptor, inhibited the hypertrophic response to CLEN. Conclusion: These data show that clenbuterol acts directly on cardiac myocytes to induce a physiological form of hypertrophy and that this is mediated, at least in part, by IGF-I. 36 TRACLEER威 (BOSENTAN), FOR THE TREATMENT OF PULMONARY ARTERIAL HYPERTENSION (PAH) – 6 MONTH QUALITY OF LIFE DATA A. Keogh, P. Macdonald, T. Williams, K. McNeil, E. Gabbay, S. Proudman, P. Steele, J. Wlodarczyk, K. Trimble, St Vincent’s Hospital, Sydney, NSW, Australia; Alfred Hospital, Melbourne, VIC, Australia; Prince Charles Hospital, Brisbane, QLD, Australia; Royal Perth Hospital, Perth, WA, Australia; Royal Adelaide Hospital, Adelaide, SA, Australia; JWCS, Sydney, NSW, Australia; Actelion Pharmaceuticals Pty Ltd, Sydney, NSW, Australia Bosentan (TRACLEER威), has demonstrated efficacy in pulmonary arterial hypertension (PAH) in placebo-controlled trials and extends the time before clinical worsening. This multicentre, open-label, single-arm Australian study was designed to assess the efficacy and safety of bosentan in patients with primary PAH (PPH) and PAH due to connective tissue disease, and to measure quality of life changes (SF36 and AQOL) with treatment. Bosentan oral therapy was commenced at 62.5mg bid for 4 weeks, increasing to a 125mg bid maintenance dose. Results: An interim analysis was conducted on the results for the 82 patients, at 6 months following start of drug. PPH represented 70%, scleroderma 25% and SLE 5%. Mean age was 51 years (range 13– 81), median PAH duration was 1.4 years (mean 3.7), and 73% of patients were female. At baseline 87% had WHO Class III symptoms and 13% Class IV. By 6 months, of 82 patients starting bosentan, 7 had died, 1 was transplanted and 17 had been withdrawn. Last outcome was carried forward for the patients withdrawn and worst case score allocated for those who died. Improvements in QOL were marked and significant on SF36 (data shown) and AQOL (not shown) in both Class III and Class IV patients.
The Journal of Heart and Lung Transplantation February 2004
Conclusion: Patients treated with open label bosentan showed very significant improvements in all aspects of quality of life on SF36 and AQOL determined after 6 months of therapy. SF36
Baseline
Month 6
p Value
Physical functioning Role physical General health Vitality Social function Role emotional Bodily pain Mental health Health Transition
27.7 16.4 30.4 34.9 49.7 48.6 60.9 64.9 3.7
37.11 31.6 33.0 42.5 57.3 59.3 61.6 64.7 2.5
0.0001 0.0001 0.0468 0.0003 0.0007 0.0182 0.0126 0.01 0.0001
37 SOLUBLE CD40 LIGAND IN PULMONARY ARTERIAL HYPERTENSION - POSSIBLE PATHOGENIC ROLE OF INTERACTION BETWEEN PLATELETS AND ENDOTHELIAL CELLS J.K. Damas,1,2 K. Otterdal,1 A. Yndestad,1 H. Aass,2 S.S. Froland,1,3 S. Simonsen,2 P. Aukrust,1,3 A.K. Andreassen,2 1Research Institute for Internal Medicine, University of Oslo, Oslo, Norway; 2 Department of Cardiology, Rikshospitalet, Oslo, Norway; 3Section of Clinical Immunology and Infectious Diseases, Rikshospitalet, Oslo, Norway Background: Inflammatory processes seem to be involved in the development and progression of pulmonary arterial hypertension (PAH). CD40 ligand (L) may induce various biological responses such as matrix degradation and thrombus formation in several inflammatory disorders. We hypothesized that CD40L also could be involved in the immunopathogenic mechanisms of PAH. Methods and Results: By comparing soluble (s) CD40L levels in 32 PAH patients and 8 controls, we found several significant differences: 1) Patients with primary pulmonary hypertension (n ⫽ 13) and secondary PAH (n ⫽ 11) had increased sCD40L levels comparing controls (p ⬍ 0.05 and p ⬍ 0.01, respectively). 2) In contrast, sCD40L levels were normal in patients with chronic thromboembolic pulmonary hypertension (n ⫽ 8) and in other PAH patients using warfarin. 3) sCD40L levels were higher in arterial (femoral artery) comparing mixed venous blood (pulmonary artery), suggesting production of CD40L in the pulmonary vasculature. 4) Platelets from patients with PAH showed higher intracellular levels, enhanced spontaneous and thrombin-stimulated release of sCD40L comparing controls (p ⬍ 0.01). Finally, in vitro studies demonstrated that sCD40L induced chemokine expression (i.e. MCP-1 and IL-8) in endothelial cells. Indeed, MCP-1 and IL-8 were elevated and correlated to pulmonary vasculature resistance in PAH (p ⬍ 0.01). Surprisingly, prostacycline enhanced MCP-1 levels in PAH patients in vivo and in CD40Lstimulated HUVEC in vitro, suggesting some unfavorable effects of this medication in PAH. Conclusion: Raised platelet-derived CD40L levels in PAH patients may (i) reflect enhanced platelet activation and thrombus formation in this disorder, and (ii) induce chemokine expression in endothelial cells. These processes could contribute to the pathogenesis of PAH. 38 A PROSPECTIVE LONGITUDINAL STUDY OF AIRWAY VASCULATURE IN THE PATHOGENESIS OF BRONCHIOITIS OBLITERANS SYNDROME(BOS) S.Y. Langenbach,2 L. Zheng,2 T. McWilliams,2 B. Levvey,1 B. Orsida,2 M. Bailey,2 T.J. Williams,1, 2 G.I. Snell,1, 2 1Department of Allergy Immunology and Respiratory Medicine, Alfred Hospital, Melbourne, VIC, Australia; 2Department of Medicine, Monash University, Melbourne, VIC, Australia