Multivariate Analysis of Radiation Responsive Proteins to Predict Radiation Exposure in Total-Body Irradiation and Partial-Body Irradiation Models

Abstract

In the event of a radiological or nuclear attack, advanced clinical countermeasures are needed for screening and medical management of the exposed population. In such a scenario, minimally invasive biomarkers that can accurately quantify radiation exposure would be useful for triage management by first responders. In this murine study, we evaluated the efficacy of a novel combination of radiation responsive proteins, Flt3 ligand (FL), serum amyloid A (SAA), matrix metalloproteinase 9 (MMP9), fibrinogen beta (FGB) and pentraxin 3 (PTX3) to predict the received dose after whole- or partial-body irradiation. Ten-week-old female C57BL6 mice received a single whole-body or partial-body dose of 18 Gy from a Pantak X-ray source at a dose rate of 2.28 Gy/min. Plasma was collected by cardiac puncture at 24, 48, 72 h and 1 week postirradiation. Plasma protein levels were determined via commercially available ELISA assay. A multivariate discriminant analysis was utilized to generate best-fit dose prediction models for whole-body exposures using the selected biomarker panel and its potential application to partial-body exposures was examined. The combination of values from FL, SAA, MMP9, FGB and PTX3 between 24 h and 1 week postirradiation yielded novel dose-response relationships. For day 1 postirradiation, the best-fit model yielded a predictive accuracy of 81% utilizing FL alone. The use of additional proteins did not enhance the model accuracy whereas, at day 2 postirradiation, the addition of PTX3 and FGB to FL increased the accuracy to 100%. At day 3 the use of FL and PTX3 yielded a predictive accuracy of 93% and at day 7 use of FL and SAA had an accuracy of 90%. Dose prediction of partial-body exposures based on the TBI model had a higher predictive accuracy when the percentage of the body exposed to radiation increased. Our findings indicate that this novel combination of radiation responsive biomarker proteins are an efficient method for predicting radiation exposure and are more accurate when used in concert compared to using any single biomarker protein alone.

Extracellular vesicles mediate low dose ionizing radiation-induced immune and inflammatory responses in the blood

“…The proteins responding to irradiation in both directly irradiated and bystander animals were acute-phase proteins (PTX-3 and lipocalin-2) (Mantovani et al 2003;Pixley and Stanley 2004;Asimakopoulou, Borkham-Kamphorst, et al 2016;, growth factors involved in stress response (M-CSF) (Pixley and Stanley 2004) or chemokines (CXCL16), indicating that cellular stress induced by ionizing radiation was transmitted by EVs to bystander mice as well. All of these proteins were already shown to be altered by radiation and various forms of oxidative stress (Haglund et al 2008;Matsumura and Demaria 2010;Shiraki et al 2012;Ariyoshi et al 2014;Tomandlova et al 2015;Isik Balci et al 2016;Yoon et al 2016;Sproull et al 2017). Two of the above four proteins were reported to be involved in radiation-induced bystander responses as well.…”

Candidate protein markers for radiation biodosimetry in the hematopoietically humanized mouse model

“…Our results (Table 2 and Supplementary Table S4 ) showed that combinations of FDXR, DDB2 and ACTN1 proteins generate an accurate prediction of the radiation dose in human lymphocytes in vivo : their mean absolute errors were ≤0.13 Gy over the tested dose range of 0 to 2 Gy. Previous studies have similarly used a combinatorial approach of plasma protein markers to provide an improved dose assessment compared to a single biomarker alone in mouse 10 and NHP models 13 . Budworth et al .…”

“…Few studies have systematically validated radio-responsive human protein markers in vivo for dose- and time-response after radiation exposure. Radiation responsive plasma proteins such as Flt3 ligand (Flt3L), serum amyloid A (SAA) and Interleukin-6 (IL-6) have been studied in mouse models as markers of acute radiation syndrome and radiation exposure up to a week 10 , 11 . Using the NHP model, C-reactive protein (CRP), SAA, IL-6, Flt3L protein biomarkers expressed in NHP plasma have been proposed as early indicators of dose assessment and radiation-induced injury up to ~7 days post irradiation 12 , 13 .…”

Biodosimetry: A Future Tool for Medical Management of Radiological Emergencies

“…Though many of the early radiation dose prediction models were based on uniform total body exposure, new research in the field is directed at developing dose prediction models for partial exposures, in addition to organ-specific markers of radiation damage. 50,51 These models are expected to be more relevant to radiological event scenarios, and identification of organ-specific markers of radiation damage will enhance medical management of radiation accident victims.…”

“…Note: Rankings are based on complexity of the model studied (in vitro, murine, non-human primate, or human clinical study), their current level of technological deployment capability, and level of advancement in the basic research. 50 operational throughput capacity, and phase of development. Cytogenetic assessment and lymphocyte kinetics/ clinical exam are rated as deployment-ready, though these technologies are not yet available at surge capacity levels.…”