Inflammation in heart failure: biomarker, bystander or mediator
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1 Inflammation in heart failure: biomarker, bystander or mediator Novel matricellular proteins to target Javier Díez, MD, PhD. Centre of Applied Medical Research and University Clinic School of Medicine, University of Navarra Pamplona, Spain cima No conflict of interest
2 MATRICELLULAR PROTEINS AND THE HEART Tenascins (C, X) Thrombospondins (1,2) Osteonectin/SPARC Osteopontin Periostin CCN1 Structural proteins Fibronectin Laminin Cell surface receptors Fibroblasts Cardiomyocytes Endothelial cells Soluble EC factors Cytokines Growth factors Proteinases Cope with environmental insults regulating inflammation, ECM remodelling, and cell survival (Schellings MWM et al, Cardiovasc Res 24;64:24-31)
3 ASSOCIATION OF OSTEOPONTIN WITH ECM IN THE HEART Experimental evidence Myocardial OPN deficiency is associated with impaired fibrotic response OPN KO mice after myocardial infarction (Circ Res 21;88:18-187) OPN KO mice submitted to ANG II infusion (Hypertension 24;43: ) OPN KO mice submitted to ANG II infusion (J Am Coll Cardiol 24;41: ) OPN KO mice submitted to aortic binding (Hypertension 24;44: ) Increased myocardial expression of OPN is associated with myocardial fibrosis Rodents with LVH of hypertensive origin (Circulation 1997;96: ) Rats after myocardial infarction (Am J Pathol 1995;145: ) Syriam hamsters with heritable DCM (Circulation 1995;92:75-79) Rats with HF due to hypertensive heart diasease (Hypertension 1999;33:663-67) Clinical evidence Increased myocardial expression of OPN is associated with myocardial fibrosis Patients with LVH of hypertensive origin (Circulation 1997;96: ) Patients with systolic HF (due to DCM of non ischemic origin) (Eur J Heart Fail 22;4: ) Patients with diastolic HF (due to HHD, HCM, and RD) (Eur J Heart Fail 22;4: )
4 OSTEOPONTIN AND MYOCARDIAL FIBROSIS Biomechanical stress or ischemic injury Mediators (ANG II, other) MAPK pathway (ERK1/2 & JNKs) Cardiomyocytes, fibroblasts, vascular cells, macrophages > Osteopontin TGF-β??? IL-1β > Myofibroblasts > Collagen formation < MMP-2, -9 Myocardial fibrosis (Singh M et al, J Mol Cell Cardiol 24;48: )
5 ROLE OF EXTRACELLUALR PROCOLLAGEN TYPE I PROTEINASES IN THE FORMATION OF COLLAGEN TYPE I MOLECULES Myofibroblast N-propeptide PCPE N-propeptide N-propeptide C-propeptide C-propeptide C-propeptide PNP N-propeptide N-propeptide N-propeptide Mature Mature Collagen Collagen type type I I Mature PCP C-propeptide C-propeptide C-propeptide PCP Procollagen type I C-terminal proteinase (or BMP-1) PCPE PCP enhancer PNP Procollagen type I N-terminal proteinase (or ADAMTS 2/14) (López B et al, Circulation 21;121: )
6 ROLE OF EXTRACELLULAR LYSYL OXIDASE IN THE FORMATION OF COLLAGEN TYPE I FIBERS Mature collagen type I Mature collagen type I Mature collagen type I LOX Fibril LOX Fiber Insoluble, high stiffness, resistant to degradation LOX Lysyl oxidase Lysine-derived cross-links (López B et al, Circulation 21;121: )
7 THE PCP/PCPE SYSTEM IN DIASTOLIC HEART FAILURE DUE TO HHD Control Diastolic HF PCP Zymogen 112 kda Mature 96 kda Controls DHF patients 1572± ±444 * 22916± ±1426 * PCPE Full length 55 kda Proteolytic fragment 36 kda.32±.3.35±.3.28±.2.39±.4 * Values (expressed as M±SEM) are given as ADU. * P<.1 vs Controls (López B et al, J Am Coll Cardiol 27;5: )
8 LOX IN DIASTOLIC HEART FAILURE DUE TO HHD Control Diastolic HF LOX 36-kDa 1.1±.9 5.2±.2 * Controls DHF patients Values (expressed as M±SEM) are given as ADU. * P<.1 vs Controls (López B et al, Hypertension 29;52: )
9 LOX IN DIASTOLIC HEART FAILURE LIKELY DUE TO HHD Parameter LOX Insoluble collagen Collagen cross-linking E -.6 (.18) -.47 (.31) -.55 (.1) E /A -.35 (.198) -.3 (.186) -.28 (.23) E/E.71 (.3).55 (.7).72 (<.1) Data are expressed as Pearson correlation coefficient (P value) (Kassner M et al, J Am Coll Cardiol 211;57: )
10
11 CARDIAC OSTEOPONTIN IN DIASTOLIC HEART FAILURE DUE TO HHD Diastolic HF Control High expression Cardiomyocytes Vascular cells Low expression Fibroblasts Macrophages (López B et al, in preparation)
12 CARDIAC OSTEOPONTIN IN DIASTOLIC HEART FAILURE DUE TO HHD Myocardial expression of osteopontin Controls DHF patients P 66 kda.61±.3.174±.8 <.5 32 kda.38±.16 * 1.52±.53 * <.5 * P<.1 vs 66kDa. Values are M±SEM. Results are given in A.U. 45. P< Osteopontin in coronary sinus blood (ng/ml) (López B et al, in preparation) Controls DHF patients
13 36 kda PCPE (A.U. x1-2 ) LOX (A.U.) Mature PCP (A.U. x1 3 ) MYOCARDIAL OSTEOPONTIN AND THE PCP/LOX AXIS IN DIASTOLIC HEART FAILURE DUE TO HHD r=.11 N.S r=.46 P< kda Osteopontin (A.U.) 14 7 r=.8 N.S kda Osteopontin (A.U.) (López B et al, in preparation)
14 MYOCARDIAL OSTEOPONTIN AND COLLAGEN AND LV STIFFNESS IN DIASTOLIC HEART FAILURE DUE TO HHD Collagen cross-lnking (mmhg/ml) Insoluble collagen (µg/mg) r=.534 P< r=.517 P< kda Osteopontin (A.U.) 32 kda Osteopontin (A.U.) LV stifness constant * r=.458 P<.5 * The LV stiffness constant was calculated from the deceleration time (AJPHCP 21;28:H554) kda Osteopontin (A.U.) (López B et al, in preparation)
15 DOES OSTEOPONTIN REGULATES CARDIAC LYSYL OXIDASE? HIF-1α AGEs + DNA OPN increases by 25% (P=.5) LOX mrna expression LOX mrna + TGF-β PGE 2 LOX precursor OPN increases by 75% (P<.1) the expression of 66 kda LOX In vitro data from experiments performed in HL-1 cardiomyocytes Procollagen C proteinase Tolloid-like 1 protein Aminopeptidase B Fibronectin + OPN does not alter the 32 kda:66 kda LOX ratio ROS LOX active form + Homocysteine Collagen cross-linking (López B et al, Am J Physiol 21;299:H1-H9) (López B et al, in preparation)
16 OSTEOPONTIN CONTRIBUTES TO MYOCARDIAL FIBROSIS VIA UPREGULATION OF LYSYL OXIDASE Biomechanical stress or ischemic injury Mediators (ANG II, other) MAPK pathway (ERK1/2 & JNKs) Cardiomyocytes, fibroblasts, vascular cells, macrophages > Osteopontin TGF-β LOX IL-1β > Myofibroblasts > Collagen formation < MMP-2, -9 Myocardial fibrosis (Singh M et al, J Mol Cell Cardiol 24;48: )
17 ACKNOWLEDGEMENT University Clinic Centre of Applied Medical Research University of Navarra & University of Navarra Pamplona, Spain Pamplona, Spain Teresa Arias, PhD Javier Beaumont, PhD Oscar Beloqui, MD, PhD José Mª. Castellano, MD, PhD Idoia Gallego, BSc María González, T Ignacio García-Bolao, MD, PhD Ana Huerta, MD Laura Martínez, BSc María Basagoiti, T Ana Egea, BSc Arantxa González, PhD Nerea Hermida, PhD Concepción Laviades, MD, PhD Begoña López, PhD Sonia Martínez, T Mika Matsumoto, MD, PhD Ramón Querejeta, MD, PhD Susana Ravassa, PhD Amaia Zudaire, BSc
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