Hydrogel delivery of purinergic enzymes improves cardiac ischemia/reperfusion injury. (March 2023)

• Record Type: Journal Article
• Authors: Sayegh, Michael N.; Cooney, Kimberly A.; Han, Woojin M.; Cicka, Markus; Strobel, Frederick; Wang, Lanfang; García, Andrés J.; Levit, Rebecca D.
• Is Part Of: Journal of molecular and cellular cardiology. Volume 176(2023)
• Page Start: 98
• Page End: 109
• Publication Date: 2023-03
• Subjects: Myocardial infarction -- Adenosine -- Ischemia -- Inflammation -- Heart failure; ATP Adenosine tri-phosphate -- ADP Adenosine di-phosphate -- AMP Adenosine mono-phosphate -- CO Cardiac output -- DAPI 4′, 6-diamidino-2-phenylindole -- EDV End diastolic volume -- ESV End systolic volume -- FITC Fluorescein isothiocyanate -- fMLP N-formylmethionyl-leucyl-phenylalanine -- GLS Global longitudinal strain -- HPLC/MS High performance liquid chromatography with tandem mass spectrometry -- HR Heart rate -- MI/R Myocardial ischemia / reperfusion -- MPO Myeloperoxidase -- NET Neutrophil extracellular trap -- PEG-4MAL Polyethylene glycol 4-maleimide -- PCI Percutaneous coronary intervention -- RGD Adhesive polypeptide -- ROS Reactive oxygen species -- SV Stroke volume -- TNFα Tumor necrosis factor alpha -- VPM Crosslinker polypeptide; Cardiology -- Periodicals; Heart Diseases -- Periodicals; Molecular Biology -- Periodicals; Cardiologie -- Périodiques; Cardiology; Electronic journals; Periodicals; 616.12
• DOI: 10.1016/j.yjmcc.2023.02.001
• Languages: English
• ISSNs: 0022-2828

Abstract

Abstract: Rationale: The innate immune response contributes to cardiac injury in myocardial ischemia/reperfusion (MI/R). Neutrophils are an important early part of the innate immune response to MI/R. Adenosine, an endogenous purine, is a known innate immune modulator and inhibitor of neutrophil activation. However, its delivery to the heart is limited by its short half-life (<30 s) and off-target side effects. CD39 and CD73 are anti-inflammatory homeostatic enzymes that can generate adenosine from phosphorylated adenosine substrate such as ATP released from injured tissue. Objective: We hypothesize that hydrogel-delivered CD39 and CD73 target the local early innate immune response, reduce neutrophil activation, and preserve cardiac function in MI/R injury. Methods and results: We engineered a poly(ethylene) glycol (PEG) hydrogel loaded with the adenosine-generating enzymes CD39 and CD73. We incubated the hydrogels with neutrophils in vitro and showed a reduction in hydrogen peroxide production using Amplex Red. We demonstrated availability of substrate for the enzymes in the myocardium in MI/R by LC/MS, and tested release kinetics from the hydrogel. On echocardiography, global longitudinal strain (GLS) was preserved in MI/R hearts treated with the loaded hydrogel. Delivery of purinergic enzymes via this synthetic hydrogel resulted in lower innate immune infiltration into the myocardium post-MI/R, decreased markers of macrophage and neutrophil activation (NETosis), and decreased leukocyte-platelet complexes in circulation. Conclusions: In a rat model of MI/R injury, CD39 and CD73 delivered via a hydrogel preserve cardiac function by modulating the innate immune response. Graphical abstract: Unlabelled Image Highlights: Hydrogel-delivered CD39 and CD73 act in tandem to convert ATP and AMP into adenosine. Purinergic hydrogels preserve cardiac function post-MI/R. Hydrogel treatment decreases leukocyte recruitment and activation. Purinergic hydrogels target neutrophil extracellular trap formation and H2 O2 generation. Platelet-leukocyte complexes in circulation are reduced in response to purinergic hydrogel treatment.