Calpain inhibition modulates glycogen synthase kinase 3β pathways in ischemic myocardium: A proteomic and mechanistic analysis

Abstract

Objectives Calpain inhibition has an enhancing effect on myocardial perfusion and improves myocardial density by up regulating by inhibition of GSK-3β and up regulating downstream signaling pathways including the insulin/PI3K and WNT/β-catenin pathways in a pig model of chronic myocardial ischemia in the setting of metabolic syndrome. Methods Pigs were fed a high fat diet for 4 weeks, then underwent placement of an ameroid constrictor to their left circumflex artery. Three weeks later animals received either: no drug (high cholesterol control: HCC), a high (HCI) or low dose (LCI) calpain inhibitor, or a GSK-3β inhibitor (GSK-3βI). The diets and CI/GSK-3βI were continued for 5 weeks and the myocardial tissue was harvested. Results Calpain and GSK-3β inhibition caused an increase in myocardial perfusion ratios at rest and during pacing compared to the control. Pigs in the LCI and HCI groups had increased vessel densities in the ischemic and pigs in the GSK-3βI group had increased vessel densities in the ischemic and non-ischemic myocardium compared to the control. Calpain inhibition modulates proteins involved in the insulin/PI3K and WNT/βcatenin pathways. Quantitative proteomics revealed that CI and GSK-3βI significantly modulated expression of proteins enriched in cytoskeletal regulation, metabolism and respiration, and calcium binding pathways. Conclusions In the setting of Metabolic Syndrome, calpain or GSK-3β inhibition increase vessel density in both the ischemic and non-ischemic myocardial tissue. Calpain inhibition may exert these effects by inhibition of GSK-3β and up regulating downstream signaling pathways including the insulin/PI3K and WNT/β-catenin pathways.

Proteomics/phosphoproteomics of left ventricular biopsies from patients with surgical coronary revascularization and pigs with coronary occlusion/reperfusion: remote ischemic preconditioning

“…Also, the phosphorylation of sarcomeric Z-disc associated proteins was increased immediately after the RIPC procedure compared to that after sham procedure in mice hearts 44 . Pharmacological interventions induced long-term expression changes of proteins related to cytoskeletal regulation in pig hearts 45 .…”

Novel molecular targets for coronary angiogenesis and ischemic heart disease

“…30,31 In a pig model of chronic myocardial ischemia in the setting of diet-induced metabolic syndrome, calpain inhibition has been shown to improve myocardial perfusion, increase endothelial dependent microvascular relaxation, decrease apoptosis and improve oxidative stress in ischemic myocardial tissue. 32–34 In the same pig model, a downstream molecular target of calpain, glycogen synthase kinase-3B, inhibition was found to have the same beneficial effects. 35 …”

Systems biology of angiogenesis signaling: Computational models and omics

“…Studies in the cardiovascular field have also started to integrate multi‐omics data (Alexandar et al, 2015; Leon‐Mimila et al, 2019; Pepin et al, 2021; Potz et al, 2017; Vilne & Schunkert, 2018). Peripheral arterial disease (PAD) is a cardiovascular disease that has a very high prevalence worldwide (Virani et al, 2021), and it is marked by narrowing or blockage of arteries in the lower limb usually due to atherosclerosis (plaque build‐up inside the vessel).…”