Metabolomic Changes in Plasma of Preterminal Stage of Rhesus Nonhuman Primates Exposed to Lethal Dose of Radiation

Abstract

Ionizing radiation exposure is known to induce molecular and cellular injury, inflicting a cascade of potentially catastrophic events leading to tissue and organ damage. Metabolomic analysis allows for the identification and quantification of small molecules downstream of genomic changes induced by radiation exposure. We aimed to characterize metabolomic changes that underscore the prefinal stage of lethally irradiated rhesus nonhuman primates (NHPs). Peripheral blood was drawn at baseline, post-exposure, as well as at the preterminal stage in NHPs (immediately prior to death in moribund NHPs) that did not survive exposure with 7.2 Gy total-body radiation (LD70/60). Herein, we analyzed global metabolomic changes using ultra-performance liquid chromatography (UPLC) quadrupole time-of-flight mass spectrometry (QTOF-MS) in plasma samples of NHPs collected at various timepoints in relation to irradiation. The overall goal was to identify metabolic shifts present immediately prior to death. Our findings showed radiation induced significant time-dependent metabolic perturbations when compared to pre-irradiation profiles, particularly in glycerophospholipid metabolism and steroid hormone biosynthesis and metabolism pathways. These findings provide valuable insights for identifying biomarkers for lethality, which may be helpful for triage during a mass casualty scenario.

Proteomic Analysis of Plasma at the Preterminal Stage of Rhesus Nonhuman Primates Exposed to a Lethal Total-Body Dose of Gamma-Radiation

“…We have also performed metabolomics analysis on plasma samples collected throughout the course of this study at the same time points, which also demonstrated that radiation induced signi cant time-dependent metabolic perturbations when compared to pre-irradiation pro les. A distinguishable preterminal phenotype was observed, with notable dysregulation in metabolites related to the glycerophospholipid metabolism and steroid hormone biosynthesis and metabolism pathways 21 . Notably, metabolomic and proteomic preterminal signatures were demonstrated in both of these studies.…”

“…These NHPs were housed in a facility accredited by the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC)-International and underwent quarantine for seven weeks. Details of animal care are described earlier 21 . The study design and animal procedures were approved by The Institutional Animal Care and Use Committee and the Department of Defense Animal Care and Use Review O ce (ACURO).…”

“…1). Plasma samples were collected pre-irradiation (day − 7), at 1, 13, and 25 days post-irradiation, and immediately prior to death in moribund animals (termed "preterminal" samples) 21 . The comparative analysis of the plasma proteomic pro les at various time points was central to this investigation, as it provides insight into the temporal dynamics of radiationinduced biological alterations.…”

“…Additionally, in an effort to address the issue of multiple comparisons that can potentially increase the likelihood of false positives, a False Discovery Rate (FDR) method was applied to adjust p-values. A more detailed summary of the statistical analyses used to analyze this data has been discussed in a recently published paper21 .Declarationsare the private views of the authors and are not necessarily those of the Uniformed Services University of the Health Sciences, or the Department of Defense. Author contributions: Study design: VKS; Performance of the study: OOF, SYW, VKS; Data acquisition, curation and analysis: VKS, AKC, ADC, OOF, SYW, JBT; Drafting of the manuscript: ADC, VKS, AKC, OOF, SYW; Revision of manuscript content: VKS, AKC, ADC, OOF, SYW; Supervision: VKS; Funding acquisition: VKS.…”

Proteomic analysis of plasma at the preterminal stage of rhesus nonhuman primates exposed to a lethal total-body dose of gamma-radiation

“…1 ). Plasma samples were collected pre-irradiation (day -7), at 1, 13, and 25 days post-irradiation, and immediately prior to death in a subgroup of moribund animals (termed “preterminal” samples) 21 . The comparative analysis of the plasma proteomic profiles at various time points was central to this investigation, as it provides insight into the temporal dynamics of radiation-induced biological alterations.…”