Page 344 - 2014 Printable Abstract Book
P. 344
(PS6-32) Serum biomarkers for detecting radiation induced lung toxicity. Feifei Song; Andrew
Branstetter; Arnab Chakravarti, MD; Naduparambil K. Jacob, PhD, The Ohio State University College of
Medicine, Columbus, OH
Approximately 50% of cancer patients receive radiation during their treatment course. Radiation
is used to target cancers, however normal tissue toxicities are inevitable and are manifested as acute
radiation syndromes in patients and as late effects in survivors. The goal of the current study is to identify
biomarkers detectable in body fluids as indicators of systemic versus organ specific response to acute
radiation exposures. Comparative and quantitative analysis of miRNAs purified from serum samples
collected from mice exposed to graded doses of radiation allowed us to develop a panel of miRNAs for
post-exposure dose estimation. An amplification-free hybridization based nCounter assay was used to
measure the time and dose dependent changes in circulating miRNAs. miR-150 was identified as a highly
sensitive biomarker whose serum depletion correlates with radiation dose after whole body exposure. In
the whole thorax and gut irradiation models, a partial depletion in miR-150 was noted, however the level
returned to baseline in three weeks, suggesting that it is an indicator of bone marrow damage and/or
recovery. Further, parallel analysis of serum levels of miRNAs in thoracic, gut versus total body irradiation
models enabled us identify organ specific responders, particularly those providing readout of lung toxicity.
Several of the miRNAs that exhibited time dependent changes in whole thorax lung irradiation models,
had high tissue expression or were reportedly connected to injury and/or inflammatory responses.
Significance of these findings in clinical biodosimetry, triage in radiation accidents and early prediction of
delayed and late effects such as pulmonary pneumonitis and fibrosis will be discussed.
(PS6-33) Development of a mass spectrometry protein biomarker panel for radiation exposure in wild-
type and NRF-2 KO mice. Andrew J. Norris, PhD; Joseph Capri, BS; Elizabeth Singer, PhD; Ewa Micewicz,
PhD; William McBride, PhD, D.Sc.; Julian P. Whitelegge, PhD; UCLA, Los Angeles, CA
The overall goal of this project is to develop a blood test for biomarkers of radiation exposure.
Our focus has been on the development and optimization of quantitative proteomics of blood plasma
from IR-exposed mice, focusing on Nrf2-modulated proteins using discovery and validation mass
spectrometry approaches. Blood plasma from wild type and Nrf2-KO mice that have received 6 Gy IR
exposure and analyzed after 2 and 5 days (compared to 0 Gy control animals) have been fractionated.
Plasma was fractionated first by pulling out the three most abundant proteins (albumin, IgG, transferrin)
and then HDL (apolipoprotein A-I and associated proteins) leaving a third ‘depleted’ fraction. Each of the
three fractions is analyzed for potential biomarkers. We present a panel of potential candidate proteins
for use as biomarkers based on their stability and persistence over 5 days. Some examples from the panel
of biomarkers include Alpha 1B Glycoprotein and Paraoxonase. Alpha 1B glycoprotein showed sustained
up-regulation wild-type plasma observed in both the depleted fraction and HDL. Paraoxonase showed a
similar trend in the HDL fraction tying radiation exposure to pro-inflammatory HDL species, and potential
up-regulation of lipid modulators of inflammation. Each protein in the panel was classified as being
involved in inflammation or apoptosis. Approximately 60% of the top 20 protein biomarkers identified in
the Nrf2-KO from the depleted fraction were involved in inflammation versus 85% in the wild type. In
addition, approximately 40% of the top 20 protein biomarkers identified in the Nrf2-KO (versus 10% in the
wild type) were involved in apoptosis. This presentation will focus upon the quantitative analysis of the
342 | P a g e
Branstetter; Arnab Chakravarti, MD; Naduparambil K. Jacob, PhD, The Ohio State University College of
Medicine, Columbus, OH
Approximately 50% of cancer patients receive radiation during their treatment course. Radiation
is used to target cancers, however normal tissue toxicities are inevitable and are manifested as acute
radiation syndromes in patients and as late effects in survivors. The goal of the current study is to identify
biomarkers detectable in body fluids as indicators of systemic versus organ specific response to acute
radiation exposures. Comparative and quantitative analysis of miRNAs purified from serum samples
collected from mice exposed to graded doses of radiation allowed us to develop a panel of miRNAs for
post-exposure dose estimation. An amplification-free hybridization based nCounter assay was used to
measure the time and dose dependent changes in circulating miRNAs. miR-150 was identified as a highly
sensitive biomarker whose serum depletion correlates with radiation dose after whole body exposure. In
the whole thorax and gut irradiation models, a partial depletion in miR-150 was noted, however the level
returned to baseline in three weeks, suggesting that it is an indicator of bone marrow damage and/or
recovery. Further, parallel analysis of serum levels of miRNAs in thoracic, gut versus total body irradiation
models enabled us identify organ specific responders, particularly those providing readout of lung toxicity.
Several of the miRNAs that exhibited time dependent changes in whole thorax lung irradiation models,
had high tissue expression or were reportedly connected to injury and/or inflammatory responses.
Significance of these findings in clinical biodosimetry, triage in radiation accidents and early prediction of
delayed and late effects such as pulmonary pneumonitis and fibrosis will be discussed.
(PS6-33) Development of a mass spectrometry protein biomarker panel for radiation exposure in wild-
type and NRF-2 KO mice. Andrew J. Norris, PhD; Joseph Capri, BS; Elizabeth Singer, PhD; Ewa Micewicz,
PhD; William McBride, PhD, D.Sc.; Julian P. Whitelegge, PhD; UCLA, Los Angeles, CA
The overall goal of this project is to develop a blood test for biomarkers of radiation exposure.
Our focus has been on the development and optimization of quantitative proteomics of blood plasma
from IR-exposed mice, focusing on Nrf2-modulated proteins using discovery and validation mass
spectrometry approaches. Blood plasma from wild type and Nrf2-KO mice that have received 6 Gy IR
exposure and analyzed after 2 and 5 days (compared to 0 Gy control animals) have been fractionated.
Plasma was fractionated first by pulling out the three most abundant proteins (albumin, IgG, transferrin)
and then HDL (apolipoprotein A-I and associated proteins) leaving a third ‘depleted’ fraction. Each of the
three fractions is analyzed for potential biomarkers. We present a panel of potential candidate proteins
for use as biomarkers based on their stability and persistence over 5 days. Some examples from the panel
of biomarkers include Alpha 1B Glycoprotein and Paraoxonase. Alpha 1B glycoprotein showed sustained
up-regulation wild-type plasma observed in both the depleted fraction and HDL. Paraoxonase showed a
similar trend in the HDL fraction tying radiation exposure to pro-inflammatory HDL species, and potential
up-regulation of lipid modulators of inflammation. Each protein in the panel was classified as being
involved in inflammation or apoptosis. Approximately 60% of the top 20 protein biomarkers identified in
the Nrf2-KO from the depleted fraction were involved in inflammation versus 85% in the wild type. In
addition, approximately 40% of the top 20 protein biomarkers identified in the Nrf2-KO (versus 10% in the
wild type) were involved in apoptosis. This presentation will focus upon the quantitative analysis of the
342 | P a g e
