André M. Leite-Moreira

Pivotal role of microRNAs in cardiac physiology and heart failure.

Cardiac hypertrophy is a hallmark of heart failure (HF), a highly prevalent, debilitating and deadly condition in Western countries. Pronounced changes in molecular pathways governing cardiac physiology underlie hypertrophy and progression to HF. MicroRNAs, small nucleotide sequences that coordinate gene expression, may have a central role in orchestrating these changes since the hypertrophic and HF myocardium clearly shows disturbed microRNA profiles. Experimental interventions targeting miR disturbances have been shown beneficial in animal models of cardiac hypertrophy and HF. This short review discusses exciting potential diagnostic and therapeutic applications of microRNAs to cardiac hypertrophy and HF, highlighting the underlying molecular pathways.

Acute Myocardial Response to Stretch: What We (don't) Know

Myocardial stretch, as result of acute hemodynamic overload, is one of the most frequent challenges to the heart and the ability of the heart to intrinsically adapt to it is essential to prevent circulatory congestion. In this review, we highlight the historical background, the currently known mechanisms, as well as the gaps in the understanding of this physiological response. The systolic adaptation to stretch is well-known for over 100 years, being dependent on an immediate increase in contractility—known as the Frank-Starling mechanism—and a further progressive increase—the slow force response. On the other hand, its diastolic counterpart remains largely unstudied. Mechanosensors are structures capable of perceiving mechanical signals and activating pathways that allow their transduction into biochemical responses. Although the connection between these structures and stretch activated pathways remains elusive, we emphasize those most likely responsible for the initiation of the acute response. Calcium-dependent pathways, including angiotensin- and endothelin-related pathways; and cGMP-dependent pathways, comprising the effects of nitric oxide and cardiac natriuretic hormones, embody downstream signaling. The ischemic setting, a paradigmatic situation of acute hemodynamic overload, is also touched upon. Despite the relevant knowledge accumulated, there is much that we still do not know. The quest for further understanding the myocardial response to acute stretch may provide new insights, not only in its physiological importance, but also in the prevention and treatment of cardiovascular diseases.

The effects of angiotensin II signaling pathway in the systolic response to acute stretch in the normal and ischemic myocardium.

Acute myocardial stretch elicits a biphasic increase in contractility: an immediate increase, known as Frank-Starling mechanism (FSM), followed by a progressive increase, regarded as slow force response (SFR). In this study, we characterized the contractile response to acute stretch from 92 to 100% Lmax in rabbit papillary muscles (n=86) under normoxic and ischemic conditions, and its modulation by angiotensin II signaling pathway. Under normoxia, the FSM was independent of Na(+)/H(+)-exchanger, reverse mode of Na(+)/Ca(2+)-exchanger (r-NCX), AT1 receptor, AT2 receptor and PKC. Regarding the SFR, it was mediated by AT1 receptor activation and its downstream effectors PKC, Na(+)/H(+)-exchanger and r-NCX. Ischemia negatively impacted on the FSM and abolished the SFR, with the muscles exhibiting a time-dependent decline in contractility. Under ischemic conditions, FSM was not influenced by AT1 and AT2 receptors or PKC activation. AT1 receptor antagonism rescued the progressive deterioration in contractility, an effect partially dependent on AT2 receptor activation.

Revisiting the slow force response: The role of the PKG signaling pathway in the normal and the ischemic heart☆

INTRODUCTION The myocardial response to acute stretch consists of a two-phase increase in contractility: an acute increase by the Frank-Starling mechanism and a gradual and time-dependent increase in force generated known as the slow force response (SFR). The SFR is actively modulated by different signaling pathways, but the role of protein kinase G (PKG) signaling is unknown. In this study we aim to characterize the role of the PKG signaling pathway in the SFR under normal and ischemic conditions. METHODS Rabbit papillary muscles were stretched from 92 to 100% of maximum length (Lmax) under basal conditions, in the absence (1) or presence of: a PKG agonist (2) and a PKG inhibitor (3); under ischemic conditions in the absence (4) or presence of: a PKG agonist (5); a nitric oxide (NO) donor (6) and a phosphodiesterase 5 (PDE5) inhibitor (7). RESULTS Under normoxia, the SFR was significantly attenuated by inhibition of PKG and remained unaltered with PKG activation. Ischemia induced a progressive decrease in myocardial contractility after stretch. Neither the PKG agonist nor the NO donor altered the myocardial response to stretch under ischemic conditions. However, the use of a PDE5 inhibitor in ischemia partially reversed the progressive deterioration in contractility. CONCLUSIONS PKG activity is essential for the SFR. During ischemia, a progressive decline in the force is observed in response to acute myocardial stretch. This dysfunctional response can be partially reversed by the use of PDE5 inhibitors.

[PP.15.22] EPINEPHRINE IS REQUIRED FOR PREVENTING BLOOD PRESSURE OVERSHOOT AND THUS CARDIAC HYPERTROPHY IN CHRONIC EXERCISE

Objective: Phenylethanolamine-N-methyltransferase-knockout (Pnmt-KO) mice are epinephrine-deficient and appear to have concentric heart remodelling. Although Pnmt-KO mice resting blood pressure is normal, it becomes higher than wild type mice during acute treadmill exercise. However, the role of epinephrine in cardiovascular response to chronic exercise remains unclear. Therefore, the aim of this study was to evaluate heart morphological, functional and molecular alterations after chronic exercise in epinephrine-deficient mice. Design and method: PCR-based genotyping was performed at the Pnmt locus of Pnmt-KO (Pnmt-/-) and wild-type (WT) mice (Pnmt+/+) (129x1/SvJ). Epinephrine and norepinephrine were quantified by RP-HPLC-ED, in adrenal glands. Animals were submitted to a 6-week chronic exercise training program performed on a motor treadmill until 20 m/min, for 55 minutes, 5 days per week. Blood pressure was determined by a photoelectric pulse detector after treadmill exercise, at rest. Mice were anaesthetized and cardiac morphology and function were evaluated by echocardiography and hemodynamics. Molecular markers of cardiac hypertrophy were evaluated by real-time PCR. Results: Systolic blood pressure was significantly increased in Pnmt-KO when compared to WT mice. A significant increase was found in left ventricular posterior wall thickness and mass in trained Pnmt-KO compared to trained WT mice, without significant differences in LV volume. Compared to basal parameters, acute &bgr;1-adrenergic stimulation with dobutamine increased cardiac index in trained WT mice, contrary to trained Pnmt-KO mice. In the left ventricle, mRNA expression of ANP and IGF-1 were significantly increased in trained Pnmt-KO mice when compared to trained WT mice. Conclusions: In conclusion, increased blood pressure overshoot in response to exercise appears to be associated with an increase in left ventricular posterior wall thickness and mass in chronic exercise, suggesting a concentric hypertrophy of the left ventricle in trained Pnmt-KO mice. In addition, acute hemodynamic stress induced by dobutamine increased systolic function index in trained WT, contrary to trained Pnmt-KO mice, suggesting a possible initial stage of pathological cardiac hypertrophy in these mice. Therefore, epinephrine appears to be essential for prevention of blood pressure overshoot and thus cardiac hypertrophy in chronic exercise.

[OP.8A.01] EXERCISE-INDUCED HYPERTENSION AND CARDIAC HYPERTROPHY IN EPINEPHRINE-DEFICIENT MICE.

Objective: Phenylethanolamine-N-methyltransferase (Pnmt) is required for the conversion of norepinephrine to epinephrine. Bao et al (2007) described that Pnmt-knockout (Pnmt-KO) mice have an increased ratio of left ventricular (LV) posterior wall thickness to internal dimensions (LVPW/LVID) but not overall cardiac hypertrophy. Pnmt-KO mice showed normal resting blood pressure (BP) but became hypertensive during treadmill exercise. The aim of this study was to evaluate cardiac morphological and functional alterations after chronic exercise in Pnmt-KO mice. Design and method: PCR-based genotyping was performed at the Pnmt locus of 10-week-old Pnmt-KO (Pnmt-/-) and WT mice (129x1/SvJ). Epinephrine and norepinephrine were quantified by RP-HPLC-ED in adrenal glands. Animals were submitted to a 6-week exercise training program. BP was measured by a photoelectric pulse detector. Mice were anesthetized (sevoflurane, 8%) and cardiac morphology and function were evaluated by echocardiography followed by morphometric analysis. Results: Epinephrine levels were vestigial in Pnmt-KO compared with WT mice. There were no differences in systolic and diastolic BP between untrained Pnmt-KO and WT mice. However, trained Pnmt-KO mice showed a significant increase in systolic BP when compared to trained WT mice. Our findings also showed that the ratios between heart weight (HW/BW) and LV weight (LVW/BW) and body weight (BW) were significantly increased in trained compared to untrained Pnmt-KO mice. Trained Pnmt-KO mice presented higher HW/BW than trained WT mice. Untrained Pnmt-KO mice display echocardiographic results similar to those reported by Bao et al (2007) while LVPW and IVS thicknesses, LV end-diastolic internal dimension (LVIDd) and LV mass indexed for body surface area (LVMi) were significantly increased in trained Pnmt-KO mice, indicating overall LV hypertrophic changes. Indexed LV end-diastolic volume (LVEDVi), indexed stroke volume (SVi) and cardiac index (CI) were significantly higher in trained Pnmt-KO than untrained Pnmt-KO mice. Conclusions: In conclusion, the increased BP in Pnmt-KO mice in response to exercise appears to be associated with an increase in LV wall thickness and chamber volume suggesting hypertrophic remodelling of the LV in Pnmt-KO mice. Epinephrine appears to be essential for maintaining normal BP, probably through &bgr;2-adrenoceptors-induced vessel relaxation, and preventing LV hypertrophy in chronic exercise.