In order to prepare for the possible detonation of a radiological dispersal device (RDD or so-called "dirty bomb"), improvised nuclear device (IND), or radiological accidents, the development of rapid, minimally invasive, and field-deployable biodosimetry is a high priority. Our project addresses this priority with the powerful global profiling capabilities of metabolomics, a biomarker discovery platform uniquely suited for the analysis of easily accessible biofluids, such as blood and urine, that require minimally- or non-invasive procedures to acquire. We have established the field of radiation metabolomics, and have published a series of seminal papers on responses at the small molecule level after radiation. Metabolomics has been used to define signatures of metabolites in urine from mice, rats, non-human primates, and humans.
After a large-scale radiological event, there will be a major need to assess, within a few days, the radiation doses received by tens or hundreds of thousands of individuals. The Center for High Throughput Minimally Invasive Radiation Biodosimetry is a research consortium to develop practical, high throughput, minimally-invasive radiation dose assessment devices and techniques to meet this need.
The Fornace lab is a member of the consortium which includes Columbia University, Georgetown University, Lovelace Repiratory Institute, NYU School of Medicine, Translational Genomics Research Institute, and the University of Bern.
The Center for High Throughput Minimally Invasive Radiation Biodosimetry is one of the Centers for Medical Countermeasures against Radiation (CMCR) supported by the National Institute of Allergy and Infectious Diseases (NIAID) through grant: U19 AI067773.
HUMAN TOTAL-BODY IRRADIATION
In a situation of a radiological accident or dirty bomb dispersal, rapid triage of potentially exposed individuals will be of importance. For this, identification of radiation biomarkers through metabolomics was expanded to a human population. Patients undergoing total body irradiation prior to hematopoietic stem cell transplantation had urine collected before and after one dose of 125cGy (6h post irradiation). Metabolomic profiling, followed by validation of the markers through tandem mass spectrometry, and quantification revealed disturbances in two major pathways, as described below.
•Developing a panel of biomarkers from human biofluids has the potential to evolve into a rapid method of identifying exposed individuals for effective triage in a situation where timely and precise diagnosis would be necessary.
In the event of a radiological event, radionuclides are feared as they will persist in the food chain and environment. Consumption of water and food will lead to internal exposure, leading to additional radiation exposure over time. We aim to understand the differences between internal and external exposure in terms of biomarker identification and metabolic pathways that are altered. The first study in collaboration with Lovelace Respiratory Research Institute focused on internal exposure to 137Cs. The study lasted for 30 days and mice received cumulative doses between 1.95 and 9.91 Gy (time dependent exposure). As highlighted in the multidimensional scaling plot, the overall metabolic profiles lead to clustering of control versus treated mice in two distinct groups. The differences between external and internal gamma exposure regarding urinary marker identification are highlighted in the table.
TARGETED METABOLOMICS AND LIPIDOMICS
- While untargeted metabolomics focuses on global changes and profiling, targeted metabolomics assesses only specific metabolites or pathways. Additional benefits include, with enrichment of samples, to quantify molecules with extremely low concentrations in biological samples. A targeted metabolomics approach was undertaken to assess changes in the serum of irradiated mice.
- The Absolute IDQTM p180 kit allowed for the identification of a large number of statistically significant ions, belonging in pathways and categories depicted in the pie chart.
- With the targeted oxylipins method we were to identify and quantify the changes in specific categories of low concentration lipids and show for the first time a significant shift from an anti-inflammatory (omega-3) to a pro-inflammatory (omega 6) phenotype following exposure to ionizing radiation (Figures 3A and 3B).
Omega 6 Proinflammatory
Omega 6 Proinflammatory
Omega 3 Anti-Inflammatory
Omega 3 Anti-Inflammatory
SELECTED COLLABORATOR PUBLICATIONS
Mukherjee, S., Laiakis, E. C., Fornace, A. J., Jr, and Amundson, S. A. Impact of inflammatory signaling on radiation biodosimetry: mouse model of inflammatory bowel disease. BMC Genomics 20: 329, 2019. PMID: 31046668
Pannkuk, E. L., Laiakis, E. C., Gill, K., Jain, S. K., Mehta, K. Y., Nishita, D., Bujold, K., Bakke, J., Gahagen, J., Authier, S., Chang, P., and Fornace, A. J., Jr. Liquid Chromatography - Mass Spectrometry based Metabolomics of Nonhuman Primates after 4 Gy Total Body Radiation Exposure: Global Effects and Targeted Panels. J Proteome Res 2019. PMID: 30843397
Laiakis, E. C., Canadell, M. P., Grilj, V., Harken, A. D., Garty, G. Y., Astarita, G., Brenner, D. J., Smilenov, L., and Fornace, A. J., Jr. Serum lipidomic analysis from mixed neutron/X-ray radiation fields reveals a hyperlipidemic and pro-inflammatory phenotype. Scientific Reports 9: 4539, 2019.
Vera, N. B., Chen, Z., Pannkuk, E., Laiakis, E. C., Fornace, A. J., Jr, Erion, D. M., Coy, S. L., Pfefferkorn, J. A., and Vouros, P. Differential Mobility Spectrometry (DMS) Reveals the Elevation of Urinary Acetylcarnitine in Non-Human Primates (NHPs) Exposed to Radiation. J Mass Spectrom 2018. PMID: 29596720 PMCID: PMC6030448
Chen, Z., Coy, S. L., Pannkuk, E. L., Laiakis, E. C., Fornace, A. J., Jr, and Vouros, P. Differential Mobility Spectrometry-Mass Spectrometry (DMS-MS) in Radiation Biodosimetry: Rapid and High-Throughput Quantitation of Multiple Radiation Biomarkers in Nonhuman Primate Urine. J Am Soc Mass Spectrom 2018. PMID: 29736597
Pannkuk, E. L., Laiakis, E. C., Fornace, A. J., Fatanmi, O. O., and Singh, V. K. A Metabolomic Serum Signature from Nonhuman Primates Treated with a Radiation Countermeasure, Gamma-tocotrienol, and Exposed to Ionizing Radiation. Health Phys 115: 3-11, 2018. PMID: 29787425 PMCID: PMC5967639
Kumar, M. A. S., Laiakis, E. C., Ghandhi, S. A., Morton, S. R., Fornace, A. J., Jr, and Amundson, S. A. Gene Expression in Parp1 Deficient Mice Exposed to a Median Lethal Dose of Gamma Rays. Radiat Res 2018. PMID: 29746213
Laiakis, E. C., Mak, T. D., Strawn, S. J., Wang, Y. W., Moon, B. H., Ake, P., and Fornace, A. J., Jr. Global metabolomic responses in urine from atm deficient mice in response to LD50/30 gamma irradiation doses. Environ Mol Mutagen 2018. PMID: 30095186
Rudqvist, N., Laiakis, E. C., Ghandhi, S. A., Kumar, S., Knotts, J. D., Chowdhury, M., Fornace, A. J., Jr, and Amundson, S. A. Global Gene Expression Response in Mouse Models of DNA Repair Deficiency after Gamma Irradiation. Radiat Res 189: 337-344, 2018. PMID: 29351057 PMCID: PMC5954417
Pannkuk, E. L., Laiakis, E. C., Garcia, M., Fornace, A. J., Jr., and Singh, V. K. Nonhuman Primates with Acute Radiation Syndrome: Results from a Global Serum Metabolomics Study after 7.2 Gy Total-Body Irradiation. Radiat Res 2018. PMID: 30183511
Pannkuk, E. L., Laiakis, E. C., Authier, S., Wong, K., and Fornace, A. J. Jr. Gas chromatography/mass spectrometry metabolomics of urine and serum from nonhuman primates exposed to ionizing radiation: Impacts on the tricarboxylic acid cycle and protein metabolism. J Proteome Res 2017. PMID: 28351153
Laiakis, E. C., Wang, Y. W., Young, E. F., Harken, A. D., Xu, Y., Smilenov, L., Garty, G. Y., Brenner, D. J., and Fornace, A. J., Jr. Metabolic Dysregulation after Neutron Exposures Expected from an Improvised Nuclear Device. Radiat Res 188: 21-34, 2017. PMID: 28475424
Laiakis, E. C., Pannkuk, E. L., Chauthe, S. K., Wang, Y. W., Lian, M., Mak, T. D., Barker, C. A., Astarita, G., and Fornace, A. J. A Serum Small Molecule Biosignature of Radiation Exposure from Total Body Irradiated Patients. J Proteome Res 2017. PMID: 28825479
Pannkuk, E. L., Laiakis, E. C., Singh, V. K., and Fornace, A. J. Lipidomic Signatures of Nonhuman Primates with Radiation-Induced Hematopoietic Syndrome. Sci Rep 7: 9777, 2017. PMID: 28852188 PMCID: PMC5575047
Goudarzi, M., Mak, T. D., Jacobs, J. P., Moon, B. H., Strawn, S. J., Braun, J., Brenner, D. J., Fornace, A. J., Jr., and Li, H. H. An Integrated Multi-Omic Approach to Assess Radiation Injury on the Host-Microbiome Axis. Radiat. Res. 186: 219-234, 2016. PMID: 27512828
Chen Z, Coy SL, Pannkuk EL, Laiakis EC, Hall AB, Fornace AJ et al. Rapid and High-Throughput Detection and Quantitation of Radiation Biomarkers in Human and Nonhuman Primates by Differential Mobility Spectrometry-Mass Spectrometry. J Am Soc Mass Spectrom 2016.
Laiakis EC, Pannkuk EL, Diaz-Rubio ME, Wang YW, Mak TD, Simbulan-Rosenthal CM et al. Implications of genotypic differences in the generation of a urinary metabolomics radiation signature. Mutat Res 2016; 788: 41-49.
Goudarzi, M., Chauthe, S., Strawn, S. J., Weber, W. M., Brenner, D. J., and Fornace, A. J. Quantitative Metabolomic Analysis of Urinary Citrulline and Calcitroic Acid in Mice after Exposure to Various Types of Ionizing Radiation. Int J Mol Sci 17: 2016. PMID: 27213362
Pannkuk EL, Laiakis EC, Authier S, Wong K, Fornace AJ Targeted metabolomics of nonhuman primate serum after exposure to ionizing radiation: potential tools for high- throughput biodosimetry. RSC Advances 2016; 6: 51192-51202.
Astarita G, Paglia G. Applications of ion mobility mass spectrometry in metabolomics. In: Advanced LC-MS Applications in Metabolomics. Borchers C, Han J, Parker C (Eds). Future Medicine, London, UK. DOI 10.4155/FSEB2013.14.63.
Goudarzi M, Weber WM, Chung J, Doyle-Eisele M, Melo DR, Mak TD, Strawn SJ, Brenner DJ, Guilmette R, Fornace AJ. Serum dyslipidemia is induced by internal exposure to strontium-90 in mice, lipidomic profiling using a data-independent LC/MS approach. J Proteome Res. 2015 Aug 11.
Laiakis EC, Trani D, Moon BH, Strawn SJ, Fornace AJ Metabolomic profiling of urine samples from mice exposed to protons reveals radiation quality and dose specific differences. Radiat Res 2015; 183: 382-390.
Goudarzi M, Weber WM, Mak TD, Chung J, Doyle-Eisele M, Melo DR, Strawn SJ, Brenner DJ, Guilmette RA, Fornace AJ Jr. A Comprehensive Metabolomic Investigation in Urine of Mice Exposed to Strontium-90. Radiat Res. 2015 May 26.
Mak TD, Laiakis EC, Goudarzi M, Fornace AJ Selective paired ion contrast analysis: a novel algorithm for analyzing postprocessed LC-MS metabolomics data possessing high experimental noise. Anal Chem 2015; 87: 3177-3186.
Paglia G, Menikarachchi LC, Langridge J, Astarita G. Travelling-wave ion mobility-mass spectrometry in metabolomics: workflows and bioinformatic tools. In: Identification and Data Processing Methods in Metabolomics. Rudaz S (Ed). Future Medicine, London, UK. DOI 10.4155/FSEB2013.14.22.
Pannkuk EL, Laiakis EC, Authier S, Wong K, Fornace AJ Global Metabolomic Identification of Long-Term Dose-Dependent Urinary Biomarkers in Nonhuman Primates Exposed to Ionizing Radiation. Radiat Res 2015; 184: 121-133.
Astarita G, Kendall AC, Dennis EA, Nicolaou A. Targeted lipidomics strategies for oxygenated metabolites of polyunsaturated fatty acids. Biochim Biophys Acta. 2014 Dec 5. pii: S1388-1981(14)00251-0. doi: 10.1016/j.bbalip.2014.11.012. [Epub ahead of print] Review.
Astarita G, McKenzie JH, Wang B, Strassburg K, Doneanu A, Johnson J, Baker A, Hankemeier T, Murphy J, Vreeken RJ, Langridge J, Kang JX. A Protective Lipidomic Biosignature Associated with a Balanced Omega-6/Omega-3 Ratio in fat-1 Transgenic Mice. PLoS One. 2014 Apr 23;9(4).
Goudarzi, M., Weber, W. M., Mak, T. D., Chung, J., Doyle-Eisele, M., Melo, D. R., Brenner, D. J., Guilmette, R. A., and Fornace, A. J. Jr. Metabolomic and lipidomic analysis of serum from mice exposed to an internal emitter, Cesium-137, using a shotgun LC-MS approach. J Proteome Res 2014. PMCID: PMC4286155
Laiakis EC, Mak TD, Anizan S, Amundson SA, Barker CA, Wolden SL et al. Development of a metabolomic radiation signature in urine from patients undergoing total body irradiation. Radiat Res 2014; 181: 350-361.
Laiakis EC, Strassburg K, Bogumil R, Lai S, Vreeken RJ, Hankemeier T, Langridge J, Plumb RS, Fornace AJ Jr, Astarita G. Metabolic Phenotyping Reveals a Lipid Mediator Response to Ionizing Radiation. J Proteome Res. 2014 Sep 5;13(9):4143-54. doi: 10.1021/pr5005295. Epub 2014 Aug 15.
Mak, T. D., Tyburski, J. B., Krausz, K. W., Kalinich, J. F., Gonzalez, F. J., and Fornace, A. J Jr. Exposure to ionizing radiation reveals global dose- and time-dependent changes in the urinary metabolome of rat. Metabolomics Epub ahead of Print, 2014.
Paglia G, Angel P, Williams JP, Richardson K, Olivos HJ, Thompson JW, Menikarachchi L, Lai S, Walsh C, Moseley A, Plumb RS, Grant DF, Palsson BO, Langridge J, Geromanos S, Astarita G. Ion Mobility-Derived Collision Cross Section As an Additional Measure for Lipid Fingerprinting and Identification. Anal Chem. 2014 Dec 29. [Epub ahead of print]
Paglia G, Williams JP, Menikarachchi L, Thompson JW, Tyldesley-Worster R, Halldórsson S, Rolfsson O, Moseley A, Grant D, Langridge J, Palsson BO, Astarita G. Ion Mobility Derived Collision Cross Sections to Support Metabolomics Applications. Anal Chem. 2014 Mar 28. [Epub ahead of print]
Sahar S, Masubuchi S, Eckel-Mahan K, Vollmer S, Galla L, Ceglia N, Masri S, Barth TK, Grimaldi B, Oluyemi O, Astarita G, Hallows WC, Piomelli D, Imhof A, Baldi P, Denu JM, Sassone-Corsi P. Circadian Control of Fatty Acid Elongation by SIRT1-mediated Deacetylation of Acetyl-CoA Synthetase 1. J Biol Chem. 2014 Jan 14. [Epub ahead of print]
Bellet MM, Nakahata Y, Boudjelal M, Watts E, Mossakowska DE, Edwards KA, Cervantes M, Astarita G, Loh C, Ellis JL, Vlasuk GP, Sassone-Corsi P. Pharmacological modulation of circadian rhythms by synthetic activators of the deacetylase SIRT1. Proc Natl Acad Sci U S A. 2013 Jan 22.
Hyduke, D. R., Laiakis, E. C., Li, H. H., and Fornace, A. J. Jr. Identifying radiation exposure biomarkers from mouse blood transcriptome. Int J Bioinform Res Appl 9: 365-385, 2013. PMCID: PMC4180524.