SWELLING ACTIVATED CURRENTS AND MYOCYTE VOLUME IN CHF

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DESCRIPTION (the applicant's description verbatim): Congestive heart failure (CHF) induces significant changes in cardiac myocyte size. Increased myocyte volume (hypertrophy) ultimately requires intracellular accumulation of osmolytes and water. Intracellular osmolarity is regulated in myocytes by multiple mechanisms, including transmembrane flux of ions through channels that are sensitive to changes in cell volume. We discovered that one of these ionic currents, the cell swelling-activated Cl- current (IC,lswell) is chronically activated under isosmotic conditions in ventricular myocytes from dogs with tachycardia-induced and rabbits with aortic regurgitation-induced CHF. Furthermore, we showed that the activity of ICl,swell and cell volume in CHF and control myocytes were regulated by protein kinase C (PKC) and protein phosphatases thought to control phosphorylation of ion channels responsible for lCl,swell. The overall objective is to understand how Icl,swell and cell volume are regulated in volume and pressure overload models of CHF and how hormonal and autocrine-paracrine factors implicated in the genesis of CHF contribute to this regulation. The effects of catecholamines, autocrine-paracrine factors including angiotensin II and cardiotrophin-1, and selected growth factors on Icl,swell and cell volume will be examined. Intracellular signaling pathways, including protein kinase C, tyrosine kinases, mitogen-activated protein kinases, and phosphatases, will be examined to evaluate their influence on lCl,swell and myocyte volume. Perforated patch voltage clamp and digital video microscopy will be used concurrently to quantify ionic currents and their effect on cell volume. Single myocytes isolated acutely from either sham operated or CHF animals will be studied because these cells better reflect the in vivo state during CHF than do cell culture models. Because no single model of CHF fully represents clinical CHF, pressure, tachycardia, and volume overload models of CHF will be used. Where appropriate, the effect of interventions on cell signaling pathways will be confirmed with western blot with phospho-antibodies. The following questions will be addressed: 1. Are lCl,swell behavior and its effect on myocyte volume different in pressure than volume overload CHF? 2. Is Icl,swell activated prior to onset of clinically apparent CHF in pressure and volume overload models? 3. Are lCl,swell and myocyte volume regulated by autocrine-paracrine factors that are important in the genesis of CHF? 4. Do intracellular signaling pathways that are important in CHF influence lCl,swell and myocyte volume? Knowledge of swelling-activated ion currents and how they influence myocyte volume in CHF may provide important insights into the pathophysiology of tachyarrhythmias and contractile and diastolic dysfunction that occur in CHF. Further, this work may lead to new approaches to treat or prevent CHF and thereby, reduce the morbidity and mortality of this common disease.