Development of Decompensated Dilated Cardiomyopathy Is Associated With Decreased Gene Expression and Activity of the Milrinone-Sensitive cAMP Phosphodiesterase PDE3A

Abstract

Background Phosphodiesterase III (PDE3) inhibitors are inotropic agents used to treat congestive heart failure (CHF) and are less effective in patients with severe CHF. Little is known about relative changes in PDE3 activity or gene expression during the evolution of cardiomyopathy. Methods and Results In the present study, we evaluated temporal changes in PDE3A gene expression before and after pacing-induced CHF in nine mongrel dogs. Three weeks of left ventricular (LV) pacing produced LV end-diastolic pressures of 15±1.7 mm Hg, whereas overt CHF at 4 to 5 weeks was associated with LV end-diastolic pressures of 24±1.7 mm Hg; prepacing values were 6.6±0.6 mm Hg. Total RNA isolated from LV tissues was analyzed on Northern blots; 10 unpaced normal hearts served as tissue controls. Signals for PDE3A mRNAs (7, 8, and 10 kb) or PDE4D (7.6 kb) were normalized against glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) or ribosomal 18S RNA. Before the onset of CHF, PDE3A / GAPDH ratios were not different between the control and 3-week paced groups. In contrast, all PDE3A / GAPDH ratios were selectively reduced by 52%, and PDE3A /18S was reduced by 70% ( P <.05) in CHF; PDE4D / GAPDH (or 18S) was unchanged. LV tissues from four control and four CHF dogs were also processed to isolate cytosolic and microsomal membrane protein for cAMP PDE3 activity assays. CHF was associated with a significant 54% reduction ( P <.05) in microsomal but not cytosolic PDE3 activity. Conclusions Selective downregulation of PDE3A may account in part for the ineffectiveness of milrinone in the treatment of severe CHF.