Activation of protein phosphatase 1 by a selective phosphatase disrupting peptide reduces sarcoplasmic reticulum Ca2+ leak in human heart failure

Abstract

Background: Disruption of Ca²⁺ homeostasis is a key pathomechanism in heart failure. CaMKII-dependent hyperphosphorylation of ryanodine receptors in the sarcoplasmic reticulum (SR) increases the arrhythmogenic SR Ca²⁺ leak and depletes SR Ca²⁺ stores. The contribution of conversely acting serine/threonine phosphatases [protein phosphatase 1 (PP1) and 2A (PP2A)] is largely unknown.

Methods and results: Human myocardium from three groups of patients was investigated: (i) healthy controls (non-failing, NF, n = 8), (ii) compensated hypertrophy (Hy, n = 16), and (iii) end-stage heart failure (HF, n = 52). Expression of PP1 was unchanged in Hy but greater in HF compared to NF while its endogenous inhibitor-1 (I-1) was markedly lower expressed in both compared to NF, suggesting increased total PP1 activity. In contrast, PP2A expression was lower in Hy and HF compared to NF. Ca²⁺ homeostasis was severely disturbed in HF compared to Hy signified by a higher SR Ca²⁺ leak, lower systolic Ca²⁺ transients as well as a decreased SR Ca²⁺ load. Inhibition of PP1/PP2A by okadaic acid increased SR Ca²⁺ load and systolic Ca²⁺ transients but severely aggravated diastolic SR Ca²⁺ leak and cellular arrhythmias in Hy. Conversely, selective activation of PP1 by a PP1-disrupting peptide (PDP3) in HF potently reduced SR Ca²⁺ leak as well as cellular arrhythmias and, importantly, did not compromise systolic Ca²⁺ release and SR Ca²⁺ load.

Conclusion: This study is the first to functionally investigate the role of PP1/PP2A for Ca²⁺ homeostasis in diseased human myocardium. Our data indicate that a modulation of phosphatase activity potently impacts Ca²⁺ cycling properties. An activation of PP1 counteracts increased kinase activity in heart failure and successfully seals the arrhythmogenic SR Ca²⁺ leak. It may thus represent a promising future antiarrhythmic therapeutic approach.

Keywords: Arrhythmia; Ca2+ cycling; Diastolic Ca2+ leak; Heart failure; Protein phosphatases.