A Quantitative Proteomic Analysis of Urine from Gamma-Irradiated Non- Human Primates

Abstract

The molecular effects of total body gamma-irradiation exposure are of critical importance as large populations of people could be exposed either by terrorists, nuclear blast, or medical therapy. In this study, we aimed to identify changes in the urine proteome using a non-human primate model system, Rhesus macaque, in order to characterize effects of acute radiation syndrome following whole body irradiation (Co-60) at 6.7 Gy and 7.4 Gy with a twelve day observation period. The urine proteome is potentially a valuable and non-invasive diagnostic for radiation exposure. Using high-resolution mass spectrometry, we identified 2346 proteins in the urine proteome. We show proteins involved in disease, cell adhesion, and metabolic pathway were significantly changed upon exposure to differing levels and durations of radiation exposure. Cell damage increased at a faster rate at 7.4 Gy compared with 6.7 Gy exposures. We report sets of proteins that are putative biomarkers of time- and dose-dependent radiation exposure. The proteomic study presented here is a comprehensive analysis of the urine proteome following radiation exposure.

Time- and radiation-dose dependent changes in the plasma proteome after total body irradiation of non-human primates: Implications for biomarker selection

“…2016 [3]. Briefly, the Rhesus monkey was selected for the study due to the fact it is a well characterized model for ARS and is the most commonly studied animal in radiation research under the US FDA Animal Rule.…”

“…The 24 Rhesus macaque plasma samples included in this study correspond directly with the previously published urine samples [3]. Venous samples were observed from a peripheral vein (i.e.…”

“…2016 [3]. The mass spectrometric raw data was analyzed by MaxQuant using a label-free intensity based approach.…”

“…Significant proteins were analyzed for pathway expression using QIAGEN’s Ingenuity Pathway Analysis (IPA, QIAGEN Redwood City, www.qiagen.com/ingenuity). Scaffold files from urine (data from [3]) and plasma proteomes were used to identify the total number of proteins found in both body fluids as well as proteins significantly differentiating all sample groups by the Analysis of Variance with normalization by iBAQ and multiple test correction by Benjamini-Hochberg using Scaffold’s Quantitative Analysis tool. Raw data used for all calculations can be found in S1 and S2 Tables.…”

“…A recent study compared the ability of plasma and urine to serve as sources of biomarkers for acute kidney injury (AKI) and concluded that the biomarkers identified in plasma had better discriminative power for preoperative risk stratification and early postoperative detection of AKI [2]. Our group previously reported a set of significantly differentiating urine proteins identified by high-resolution mass spectrometry from a study of twenty-four Rhesus macaque animals exposed to two different levels of gamma-irradiation (Co-60 at 6.7 Gy and 7.4 Gy) with a twelve day observation period [3]. In the current study, we examined the plasma from the same animals and identified a series of complementary biomarkers of radiation exposure.…”