1. Characterization of cardiac stress (Takotsubo Cardiomyopathy)-related miRNAs ex vivo Anselm Derda, ESC Congress 2015 NHLI – Prof. Sian Harding IMTTS – Prof. Thomas Thum

2. 1. 2. 3. Isolation of LV rat myocytes Transfection & Incubation for 48h   Control/Inhibitor/Mimic of miR-16 Contractility/IonOptix Basal contractility

3.  Precursor of miR-16 reduced contractility  Inhibitor of miR-16 increased contractility N(rat) = 5 MEAN + SEM N(rat) = 5 MEAN + SEM p < 0.0001 p = 0.0194 17 22 16 Basal contractility

4. 1. 2. 3. Isolation of LV rat myocytes Transfection & Incubation for 48h   Control/Inhibitor/Mimic of miR-16   add Pertussis Toxin Contractility/IonOptix + PTX Contractility after block of Gi

5.  PTX removed decreasing effect of mimic of miR-16 N(rat) = 4 MEAN + SEM p = 0.53 15 20 Contractility after block of Gi

6. 1. 2. 3. Isolation of LV rat myocytes Transfection & Incubation for 48h   Control/Inhibitor/Mimic of miR-16 Contractility/IonOptix Contractility with isoprenaline simulating stress + isoprenaline β 1 β 2 + CGP

7. time length isoprenaline CGP max response min response Schematic: isoprenaline response control Inhibitor miR-16 max response min response = =  Inhibitor of miR-16 did not affect positive inotropic isoprenaline response

8. Conclusion -miR-16 had an Gi-dependent negative inotropic effect on cardiomyocytes -miR-16 did not affect the contractility of cardiomyocytes under stress conditions induced by isoprenaline -  miR-16 is possibly influencing the tonic effect of Gi on contraction -Further experiments are necessary for... -...evaluating signal cascade of miR-16 more precisely -...showing a possible interaction between miR-16 and adenylate cyclase