Data-Independent Acquisition Proteomics Reveals Long-Term Biomarkers in the Serum of C57BL/6J Mice Following Local High-Dose Heart Irradiation

Abstract

Background and Purpose: Cardiotoxicity is a well-known adverse effect of radiation therapy. Measurable abnormalities in the heart function indicate advanced and often irreversible heart damage. Therefore, early detection of cardiac toxicity is necessary to delay and alleviate the development of the disease. The present study investigated long-term serum proteome alterations following local heart irradiation using a mouse model with the aim to detect biomarkers of radiation-induced cardiac toxicity.Materials and Methods: Serum samples from C57BL/6J mice were collected 20 weeks after local heart irradiation with 8 or 16 Gy X-ray; the controls were sham-irradiated. The samples were analyzed by quantitative proteomics based on data-independent acquisition mass spectrometry. The proteomics data were further investigated using bioinformatics and ELISA.Results: The analysis showed radiation-induced changes in the level of several serum proteins involved in the acute phase response, inflammation, and cholesterol metabolism. We found significantly enhanced expression of proinflammatory cytokines (TNF-α, TGF-β, IL-1, and IL-6) in the serum of the irradiated mice. The level of free fatty acids, total cholesterol, low-density lipoprotein (LDL), and oxidized LDL was increased, whereas that of high-density lipoprotein was decreased by irradiation.Conclusions: This study provides information on systemic effects of heart irradiation. It elucidates a radiation fingerprint in the serum that may be used to elucidate adverse cardiac effects after radiation therapy.

Activation of PPARα by Fenofibrate Attenuates the Effect of Local Heart High Dose Irradiation on the Mouse Cardiac Proteome

“…The levels of seven different cytokines and inflammatory mediators were measured in the serum of mice using ELISA. In good agreement with our previous data [ 28 ], the levels of TGF-β, IL-1 α and β, and IL-6, were significantly increased following irradiation (16 Gy) in comparison to the controls ( Figure 6 B). Importantly, when using fenofibrate treatment the levels of pro-inflammatory cytokines were restored to the levels observed in controls ( Figure 6 B).…”

“…Furthermore, the serum of mice was tested for levels of free fatty acids (FFA). In good agreement with previous data [ 11 , 28 ], the level of free fatty acids was increased in the serum after local heart exposure (16 Gy) ( Figure 3 E) suggesting a radiation-induced impairment of the fatty acid metabolism. In the fenofibrate-fed mice, the amount of serum FFA returned to normal levels ( Figure 3 E).…”

“…The impairment of the PPARα signalling pathway, as observed in irradiated hearts, might be involved in the alteration of the cardiac oxidation/reduction equilibrium and may result in chronic inflammation [ 67 ]. Our data showed that the radiation-induced increase in the inflammatory response [ 28 , 37 , 72 , 73 ] was prevented in fenofibrate-treated mice. The activation of PPARα has been shown to reduce the production of several inflammatory genes or proteins including TNF-α, IL-1β and IL-6, suggesting a reduction in the systemic inflammatory response [ 17 , 18 , 74 ].…”

“…In total, 20 mice were used in this study, with 5 mice in each group. Male animals were used so the results of this study could be compared with results from our earlier studies [ 11 , 12 , 13 , 28 ].…”

A Novel Classifier Based on Urinary Proteomics for Distinguishing Between Benign and Malignant Ovarian Tumors

“…Proteomics based on mass spectrometry (MS) is a powerful technique increasingly employed not only for high-throughput identification but also quantification of multiple proteins. Dataindependent acquisition (DIA) MS has recently emerged as a promising alternative to data-dependent acquisition (DDA) for quantitative proteomics analysis (Azimzadeh et al, 2021;Prestagiacomo et al, 2021). The DIA technique is widely used in the context of multiplex biomarker detection from clinical specimens, such as plasma and urine (Fang et al, 2020;Burnap et al, 2021).…”