New drugs on the horizon for heart failure: CaMK antagonists

Transcription

1 New drugs on the horizon for heart failure: CaMK antagonists (Lars S. Maier) Gerd Hasenfuss Disclosure Information: No relationship exists related to this presentation Dept. of Cardiology and Pneumology, Heart Center Heart Research Center Göttingen, Germany

2 Maier & Bers, JMCC, 22 Maier et al., JMCC, 26 Ca/calmodulin-dependent kinase II (CaMKII) multifunctional serine/threonine protein kinase isoform predominant in heart ( C cytosol, B nucleus) N regulatory domain Thr 287 catalytic domain Inactive N C Target N ATP catalytic domain Thr 287 Direct activation Target N ATP catalytic domain Thr 287 P Autonomous (2-8% active)

3 Currently available CaMKII inhibitors KN-93 (2-[N-(2-Hydroxyethyl)]-N-(4- methoxybenzenesulfonyl)]amino-n-(4- chlorocinnamyl)-n-methylbenzylamine) AIP (Autocamtide 2-related inhibitory peptide) MW 599 MW 1498 IC 5 1 µm C 26 H 29 ClN 2 O 4 S H 3 PO 4 IC 5 1 nm Lys-Lys-Ala-Leu-Arg-Arg-Gln-Glu-Ala-Val- Asp-Ala-Leu N regulatory domain Thr 287 catalytic domain Inactive N Target N KN93 C ATP AIP catalytic domain Thr 287 Direct activation Target N ATP catalytic domain Thr 287 P Autonomous (2-8% active)

4 T-Tubule RyR AP Na 3Na Sarcolemma NCX Na P P Ca I Ca P P SR PLB SERCA P P Ca Ca Myofilaments NCX 3Na Maier & Bers, Cardiovasc Res, 27

5 8 nmol/l CaMKII C overexpression in TG mice leads to heart failure, survival & SR Ca content mouse: TG CaMKII C Percent survival (%) TGCaMKII C Age (weeks) Caffeine 1 mmol/l TG SERCA: NCX: TG 5 s Zhang et al., Circ Res, 23 Maier et al., Circ Res, 23

6 CaMKII-dependent SR Ca leak A TG TG CaSpF (pl -1 s -1 ) mouse: CaMKII 1 m TG upregulation in HF Hoch et al Circ Res F/F 1 1 ms 2 B 1 F (a.u.) 1 ms 1 m Control KN-93 CaMKII inhibition Diastolic SR Ca leak (spark frequency amplitude duration width) is 4.3 times in TG vs.! Maier et al., Circ Res, 23 Kohlhaas et al., Circ Res, 26

7 mouse: CaMKII expression and hypertrophy TAC (afterload) SHUNT (preload) CaMKII B CaMKII C 7 days Survival also HDAC-P PKD P-CaMK/GAPDH P<.5 Percent survival 1 Sham 75 5 Shunt TAC 25 # Sham TAC Sham Shunt Days Toischer et al., Circulation, 21

8 Apoptosis in CaMKIIδ-KO 7 days after TAC cells per mm activated caspase3 P<.1 P<.1 P<.1. Sham TAC Sham TAC KO

9 CaMKII-KO reduces cardiac hypertrophy CaMKII is activated in Hypertrophy and heart failure (with increased afterload) Significant contributer to pathophysiology Backs et al., PNAS, 29

10 EADs and DADs in TG CaMKII C mice mouse: 1 mv 25 ms 3 1 mv 25 ms TG CaMKII 5 mv 3 25 ms TG M ISO EADs (per 1 APs) 2 1, N=25 P<.5 TG, N=31 P<.5 TG + AIP, N=7 DADs and spontaneous APs (per 1 APs) 2 1 P<.5, N=5 TG, N=6 Sag et al., Circ Heart Fail, 29

11 Arrhythmias (%) Arrhythmias can be elicited in TG CaMKII C mice in vivo mouse: no arrhythmia Isoproterenol.25 mv 4 ms 12 1 TG 8.25 mv Monomorphic VT mv TG Polymorphic VT 2 /7 6/7 TG Wagner et al., JCI, 26

12 CaMKII inhibition proarrhythmogenic mouse: (non-stimulated) events in vitro single events sustained events NSE NSE stim late NSE (% of cells) TG control, n= 5 KN-92, n=42 KN-93, n=46 sustained NSE [% cells] n = 31 TG n = 33 TG + KN-93 n = 8 Sag et al., Circ Heart Fail, 29

13 1 1 mv 1 mv 1 mv CaMKII inhibition propensity for arrhythmias in CaMKII C TG mice in vivo mouse: ISO ISO-induced arrhythmias 5 s TG CaMKII C P=N.S. P<.5 ISO 5 s ISO 5 s ISO-induced arrhythmias +KN-92 (2µM/kg/KG) +KN-93 (2µM/kg/KG) Arrhythmias (in %) TG CaMKII C, n=5 KN-92, n=4 KN-93, n=4 Sag et al., Circ Heart Fail, 29

14 CaMKII in human heart failure Kirchhefer et al., Cardiovasc Res, 1999 Hoch et al., Circ Res, 1999

15 CaMKII expression also during LV hypertrophy in patients with aortic stenosis human: LV AKE vs. NF Ratio CaMKII/GAPDH Nonfailing Hypertrophy CaMKIIδ B CaMKIIδ C GAPDH H 1 NF 1 H 2 NF 2 H 3 NF 3 58 kda 56 kda 37 kda Toischer et al., Circulation, 21

16 CaSpF (pl -1 s -1 ) AIP 1µm 1µm CaMKII inhibition SR Ca leak in heart failure human: Basal 2 ms AIP 2 ms Control SR-Ca 2+ -leak (pf/f /µm 2 ) Control AIP Sossalla et al., Circ Res, 21

17 CaMKII inhibition contractility and SR Ca content in heart failure human:.5 Hz 1 Hz 2 Hz 3 Hz 18 KN Force (mn/mm²) Force (mn/mm²) Force (mn/mm²) Force (mn/mm²) time (ms) time (ms) time (ms) time (ms) KN Force (mn/mm²) Force (mn/mm²) Force (mn/mm²) Force (mn/mm²) time (ms) time (ms) time (ms) SR Ca content time (ms) F/F 1.5 KN KN Time (ms) Sossalla et al., Circ Res, 21

18 SR Ca leak & diastolic [Ca] during AF human: Sinus rhythm KN-92 Sinus rhythm KN-93 1 µm 2 ms Atrial fibrillation KN-92 Atrial fibrillation KN-93 calc. total SR Ca 2+ leak (nmol/l) KN-92 KN-93 Sinus rhythm n.s. KN-92 KN-93 Atrial fibrillation diastolic Ca 2+ (nmol/l) KN-92 KN-93 Sinus rhythm KN-92 KN-93 Atrial fibrillation Neef et al., Circ Res, 21

19 CaMK is activated in pressure overload and heart failure disturbed Ca cycling contractile dysfunction arrhyhtmias apoptosis and death prevention by CaMK-KO or CaMK inhibitors CaMK is promising drug target in heart failure but it is everywhere

20 Thanks to K. Toischer, Göttingen Don Bers, UCD Joan Heller Brown, UCSD Johannes Backs, Heidelberg Eric Olson, Dallas Sebastian Maier, Würzburg DFG (Emmy-Noether-Program, Heisenberg-Program, KFO) DGK (Hengstberger Stipend) GSK Research Foundation CVT/Gilead EU-Projects: EuGeneHeart, CONTICA Transatlantic Leducq networks CaMKII and ROS

21 TG CaMKII inhibition using AC3-I protects against structural heart disease mouse: N Target N KN93 C AC3-I AIP ATP catalytic domain Thr 287 Direct activation AC3-I overexpression (I=CaMKII inhibition) using the cardiac-specific Myh6 promoter vs. the inactive scrambled control peptide (AC3-C, C=control) Zhang et al.,, Nat Med, 25