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Florindi, C.

Publications and source records attributed to Florindi, C..

2 recordsLinked to original sources

Cellular consequences of non-ablative radiotherapy, a novel approach to ventricular tachycardias

BackgroundRadiotherapy (RT) with a single focused application of ionizing radiation (STAR) has been suggested as a non-invasive alternative to radiofrequency in ablating ventricular tachycardia (VT). Emerging data reveal that STAR may suppress VT without substrate destruction, by enhancing impulse conduction instead, through increased expression of NaV1.5 channels and connexin 43. AimsTo investigate electrophysiology and intracellular Ca2+ dynamics in cardiomyocytes (CMs) from mice subjected to in-vivo RT. The emerging data led us to evaluate biochemical changes potentially linking electrophysiological response to ionizing irradiation. MethodsCMs isolated 2 weeks after RT with low-dose (15 Gy) or high-dose (25 Gy) were compared to those of sham-treated mice (CTRL). We evaluated: i) INaT and INasus properties; ii) AP parameters, including the prevalence of Early After-Depolarizations (EADs); iii) intracellular Ca2+ dynamics; iv) CaMKII phosphorylation and v) ROS content. Results25 Gy RT i) increased INaT and, to a larger extent, INasus (increased INasus/INaT ratio); ii) increased AP amplitude, +dV/dtmax and duration (APD) and facilitated EADs; iii) depressed intracellular Ca2+ dynamics. 15 Gy RT had similar but smaller effects (dose-dependency). 25 Gy RT reduced CaMKII phosphorylation but increased cell ROS content, thus providing a mechanism for INaL enhancement. ConclusionsThe results support the view that STAR may supress VT by increasing conduction velocity, with APD prolongation providing an additional mechanism. On the other hand, INasus enhancement (likely by ROS) and Ca2+ handling depression may impair electrical stability and contractility in the irradiated region.

physiology↗

Early consequences of the phospholamban mutation PLN-R14del+/- in a transgenic mouse model

AimsThe heterozygous phospholamban (PLN) mutation R14del (PLN R14del+/-) is associated with a severe arrhythmogenic cardiomyopathy (ACM) developing in the adult. "Superinhibition" of SERCA2a by PLN R14del is widely assumed to underlie the pathogenesis, but alternative mechanisms such abnormal energy metabolism have also been reported. This work aims to 1) to evaluate Ca2+ dynamics and energy metabolism in a transgenic (TG) mouse model of the mutation prior to cardiomyopathy development; 2) to test whether they are causally connected. Methods and ResultsCa2+ dynamics, energy metabolism parameters, reporters of mitochondrial integrity, energy and redox homeostasis were measured in ventricular myocytes of 8-12 weeks-old, phenotypically silent, TG mice. Mutation effects were compared to pharmacological PLN antagonism and analysed during modulation of sarcoplasmic reticulum (SR) and cytosolic Ca2+ compartments. Transcripts and proteins of relevant signalling pathways were evaluated. The mutation was characterized by hyperdynamic Ca2+ handling, similar to that induced by PLN antagonism. Albeit all components of energy metabolism were depressed at rest, functional signs of mitochondrial damage or energy starvation were absent and cell energy charge was preserved. The response of mitochondrial O2 consumption to SERCA2a blockade was lost in mutant myocytes (SR-mitochondrial uncoupling) and ER-stress signalling was activated. Conclusions1) PLN R14del+/- loses its ability to inhibit SERCA2a, which argues against SERCA2a superinhibition as a mechanism of ACM; 2) depression of resting energy metabolism may at least partly reflect impairment of SR-mitochondrial coupling; 3) ER-stress may be an early factor in the pathogenesis.

physiology↗