Andrew Remppis

Therapeutic safety of high myocardial expression levels of the molecular inotrope S100A1 in a preclinical heart failure model

Low levels of the molecular inotrope S100A1 are sufficient to rescue post-ischemic heart failure (HF). As a prerequisite to clinical application and to determine the safety of myocardial S100A1 DNA-based therapy, we investigated the effects of high myocardial S100A1 expression levels on the cardiac contractile function and occurrence of arrhythmia in a preclinical large animal HF model. At 2 weeks after myocardial infarction domestic pigs presented significant left ventricular (LV) contractile dysfunction. Retrograde application of AAV6-S100A1 (1.5 × 1013 tvp) via the anterior cardiac vein (ACV) resulted in high-level myocardial S100A1 protein peak expression of up to 95-fold above control. At 14 weeks, pigs with high-level myocardial S100A1 protein overexpression did not show abnormalities in the electrocardiogram. Electrophysiological right ventricular stimulation ruled out an increased susceptibility to monomorphic ventricular arrhythmia. High-level S100A1 protein overexpression in the LV myocardium resulted in a significant increase in LV ejection fraction (LVEF), albeit to a lesser extent than previously reported with low S100A1 protein overexpression. Cardiac remodeling was, however, equally reversed. High myocardial S100A1 protein overexpression neither increases the occurrence of cardiac arrhythmia nor causes detrimental effects on myocardial contractile function in vivo. In contrast, this study demonstrates a broad therapeutic range of S100A1 gene therapy in post-ischemic HF using a preclinical large animal model.

Purification of the Ca2+-binding protein S100A1 from myocardium and recombinant Escherichia coli

S100A1 is a new regulatory protein of myocardial contractility that is differentially expressed in early and late stages of myocardial hypertrophy. In order to further investigate the multiple functions of S100A1 in various assay systems we developed a new strategy for isolating biologically active S100A1 protein. After EDTA extraction of myocardium or recombinant bacteria, S100A1 was purified by Octyl-Sepharose hydrophobic interaction chromatography and HiTrapQ anion-exchange chromatography yielding 1.4-2.0 mg/100 g wet tissue and 0.7-1.0 mg/100 ml bacterial culture. Native porcine as well as human recombinant S100A1 revealed biological activity in physiological and biochemical assays.

Wertigkeit myofibrillärer Proteine in der Diagnostik des akuten Myokardinfarktes

Die Diagnose eines akuten Myokardinfarktes kann bei Patienten mit typischer pektanginoser Symptomatik und signifikanten monophasischen ST-Hebungen im EKG mit so hoher Sicherheit gestellt werden, das eine Bestatigung der Diagnose durch laborchemische Methoden vor Einleitung einer thrombolytischen Therapie nicht erforderlich ist (Yusuf et al. 1984). Dagegen hangt die Diagnose kleiner Myokardinfarkte oder nichtischamischer Herzmuskelnekrosen entscheident von der Effizienz laborchemischer Verfahren ab.