Quantitative Proteomics for Cardiac Biomarker Discovery Using Isoproterenol-Treated Nonhuman Primates

Abstract

To identify new cardiac biomarkers, a quantitative proteomic analysis has been performed on serum and heart tissue proteins from three species of nonhuman primates following isoproterenol (ISO) treatment. Three serum proteins—serum amyloid A (SAA), α-1-acid glycoprotein (A1AG), and apolipoprotein A-1 (Apo A1)—were consistently identified as changed and remained altered 72 h post dose in all three species post ISO treatment, indicating the potential of including these proteins in preclinical or clinical evaluation of drug-induced cardiac injury. Furthermore, proteomic analysis of heart tissue proteins following ISO treatment demonstrated detrimental effects on calcium signaling and energy generation in cardiac myocytes. It is worth noting that cardiac troponins were not identified in serum but were identified as altered in heart tissue lysate along with other cardiac-specific proteins. This strategy for cardiac biomarker discovery by proteomic screening of heart tissue proteins, followed by verification in serum samples using immunoassays or targeted mass spectrometry, could be applied in future biomarker studies.

Unveiling the molecular crosstalk in a human induced pluripotent stem cell‐derived cardiac model

“…Drug cardiotoxicity assays using human cell‐based in vitro models often rely on viability readouts obtained using spectrophotometric, fluorescent, or imaging techniques, allowing for high‐throughput but not in‐depth characterization of the molecular mechanisms occurring in cardiotoxicity. Only recently, proteomics‐driven approaches started to play a role in the characterization of cardiac molecular toxicological mechanisms and on the identification of biomarkers induced by different drugs (Song et al, ; Štěrba et al, ). These studies have been performed in animal models but not in cardiac tissues engineered in vitro.…”

Analysis of the plasma proteome using iTRAQ and TMT‐based Isobaric labeling

“…Isobaric labeling analyses of plasma samples have also been made without immunodepletion or ligand affinity, including direct labeling of undepleted plasma, as well as other modes of fractionation followed by labeling. For example, undepleted serum was TMT labeled, followed by on‐line fractionation in a study of biomarkers of cardiac injury . In another depletion‐free approach, a three‐dimensional fractionation strategy was used.…”

“…These have included atherosclerotic disease, abdominal and thoracic aneurysms, arterial and pulmonary embolisms, aortic dissection haemorrhagic shock and hypertrophic cardiomyopathy . Investigating markers of cardiac injury, Liu and Song used an ape model together with isoproterenol treatment …”

“…32 37,38 There have, however, been some indications that the relative capture efficiencies of the hexapeptide beads can vary as much as two-fold with some proteins. 37 Isobaric labeling analyses of plasma samples have also been made without immunodepletion or ligand affinity, [39][40][41] including direct labeling of undepleted plasma, as well as other modes of fractionation followed by labeling. For example, undepleted serum was TMT labeled, followed by on-line fractionation in a study of biomarkers of cardiac injury.…”

Evaluation of Cardiac Toxicity Biomarkers in Rats from Different Laboratories

“…Various studies have been conducted to evaluate myocyte damage following exposure to this drug (York et al , 2007, Kaufman, 1985, Hasic et al , 2011, Acikel et al , 2005, Brady et al , 2010). We have previously reported on a variety of serum proteins showing significant changes in African green monkeys, Rhesus macaques and Cynomolgus macaques; these proteins can potentially be used for predicting long-term cardiac injury (Liu et al , 2013, Song et al , 2014), but are not currently available in standardized kits.…”