Gulf War Illness Research Program

Gulf War illness (GWI) is a multi-symptom disorder affecting one fourth of military veterans of the First Persian Gulf War (1990-91). GWI veterans have an extended spectrum of complaints including fatigue, cognitive impairments, diffuse musculoskeletal pain, gastrointestinal dysfunction and sleep abnormalities. The heterogeneity and complexity of GWI has led to difficulties in diagnosis and treatment.

Roskamp Institute successfully explores the effects of chemical agents from the Gulf War, including pyridostigmine bromide (PB) and the pesticide permethrin (PER), developing novel mouse models of GWI. Studies include characterization of the cognitive and neurobehavioral impairment in the animals, as well as and analysis of blood and brain samples.

Furthermore, Roskamp Institute has a well-established biomarker research program, which is particularly focused on translating animal modeling work to clinical application. Roskamp’s solution to the problem of a lack of peripheral biomarkers for GWI diagnosis and clinical management is to apply our expertise in preclinical animal modeling of CNS-based disorders to the generation of plasma biomarker profiles from mouse models of GW agent-exposed mice which display key CNS features relevant to veterans with GWI. Through collaboration with James J. Peters VA Medical Center, Bronx, NY and Boston VA Medical Center, Boston, MA, Roskamp Institute is able to use Omics technology in human clinical populations of Gulf War Veterans. This allows us to translate preclinical findings to human populations and develop biomarker panels for the diagnosis and treatment of GWI.

GWI Research Team

Fiona Crawford, Ph.D. –
Director, GWI Research Program
Laila Abdullah, Ph.D.
Ghania Ait-Ghezala, Ph.D.
Tanja Emmerich, M.S., Ph.D. student
Zuchra Zakirova, M.S., Ph.D. student
Thinh Nguyen, B.S. – Research Assistant

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Laboratory Models for GWI
The Roskamp Institute GWI Research Program is dedicated to finding therapeutics that can target the central nervous system (CNS) pathology of GWI. The scientific research team has developed and characterized several mouse models of Gulf War (GW agent) exposure using the acetylcholinesterase inhibitor pyridostigmine bromide (PB), the insect repellent N,N-diethyl-meta-toluamide (DEET) and pesticides such as permethrin (PER, a pyrethroid) and chlorpyrifos (CPF, an organophosphate). These chemicals have been implicated as potential causal factors in GWI. These GWI mouse models exhibit neurobehavioral deficits, including anxiety and memory problems that are similar to the symptoms reported by veterans with GWI. These mouse models exhibit increased astroglial activation in the brains of exposed mice, which is now thought to be a key CNS feature of GWI observed in laboratory models of GWI. The Roskamp Institute research team is now focused on characterizing their primary mouse model that uses combined PB+PER exposure. This PB+PER mouse model exhibits cognitive impairment and astroglial pathology even at 16- and 22-months post-exposure. This is highly relevant to GWI since a large proportion of veterans from the 1991 Gulf War have been experiencing persistent health problems for over twenty years. Currently, a detailed molecular characterization of this model is underway using advance omic technologies (proteomic and lipidomics). It is anticipated that the mouse models of GWI developed by the Roskamp Institute GWI research team will be invaluable in guiding translational efforts at the Institute that are aimed at identifying appropriate therapies for treating GWI.

Lipidomics
Using proteomics, RI scientists identified that lipid metabolism, immune/inflammatory and endocrine functions were affected in the brains of PB+PER exposed mice. Lipidomics studies revealed that lipid dysfunction in the brains of exposed mice was present at several chronic post-exposure time-points. Given that lipid metabolism can influence many biological functions, including immune/inflammation and endocrine functions, targeting lipid metabolism may be a new approach by which several features of GWI may be improved.

Dr. Laila Abdullah’s lipidomic research at RI is currently focused on identifying biological functions and pathways that are associated with abnormal lipid profiles in mouse models of GWI. She is using novel mass spectrometry and standard molecular biology techniques to determine if cellular functions that are involved in lipid synthesis, clearance and metabolism are affected by PB+PER exposure. Since astroglia cells in the brain play an important role in lipid transport and metabolism, she will conduct additional studies to better characterize the relationship between dysfunctional lipid presentation and astroglia changes that are seen in this mouse model of GWI. As part of these studies, she will identify treatments that target lipid dysfunction seen in mouse model of GWI. She will test natural compounds or FDA approved drugs as therapies that target lipid dysfunction in these preclinical studies to minimize the risk of translational failures and ensure a better success rate in developing treatments for veterans with GWI. Currently, there are no therapies available that can provide disease modification of the underlying CNS pathology associated with GWI. Dr. Abdullah will use exceptional resources and expertise available at the Roskamp Institute to identify the underlying pathobiology of chronic CNS symptoms of GWI, which will yield potential therapies for treating the chronic CNS symptoms of GWI.

Proteomics
Proteomics is a field of bioanalytical chemistry that focuses on the characterization of proteins. This technology enables researches to examine thousands of proteins from complex samples with state-of-the-art instrumentation. Using this approach, it is possible to identify changes in certain proteins between control and diseased populations. These changes are often quantifiable, and can be used to determine the degree to which a protein is changing. This information is then used to identify and compare the biochemical mechanisms at work in normal and diseased states, or to identify specific markers (biomarkers) of the disease â€“ or in many cases, both.

Dr. Ait-Ghezala’s team at the Roskamp Institute (RI) centers on examining brain tissue from animals that have been exposed to a combination of pyridostigmine brominde (PB), an acetylcholinesterase inhibitor, and permethrin (PER), a pesticide, â€“ two of the primary agents believed to be responsible for the underlying the pathobiology of GWI. Using the proteomic techniques TMT and iTRAQ (Tandem Mass Tagging, and Isobaric Tags for Relative and Absolute Quantification, respectively), Dr. Ait-Ghezala and her team have identified proteins involved in several canonical pathways, including Mitochondrial Dysfunction, Fatty Acid metabolism, and Immune/ Inflammatory responses. These findings are consistent with previous work involving veterans with GWI, and shed new light on possible mechanisms regarding the pathobiology of this chronic multi-symptom illness.

Biomarkers
Roskamp Institute has a well-established biomarker research program, which is particularly focused on translating animal modeling work to clinical application. Roskamp’s solution to the problem of a lack of peripheral biomarkers for GWI diagnosis and clinical management is to apply our expertise in preclinical animal modeling of CNS-based disorders to the generation of plasma biomarker profiles from mouse models of GW agent-exposed mice which display key CNS features relevant to veterans with GWI. Through collaboration with James J. Peters VA Medical Center, Bronx, NY and Boston VA Medical Center, Boston, MA, Roskamp Institute is able to use Omics technology in human clinical populations of Gulf War Veterans. This allows us to translate preclinical findings to human populations and develop biomarker panels for the diagnosis and treatment of GWI.

Mitochondrial Dysfunction / Neuroinflammation
Dr. Ait-Ghezala, has evaluated neuropathology and cognitive impairment, and assayed the expression of thousands of proteins in the PB+PER exposure model using an unbiased proteomic approach. She applied bioinformatics analysis to identify specific protein changes and biomolecular pathways impacted by exposure. This work has identified several targets for potential therapeutic intervention in GWI, namely neuroinflammation and mitochondrial dysfunction. She demonstrated that there is a chronic inflammatory CNS and peripheral disturbance in the brains of exposed versus non-exposed mice. Dr. Ait-Ghezala also found evidence of an inflammatory response that is accompanied by a profound increase in astrogliosis, which is a consistent feature at chronic time-points post-exposure in every cohort of mice she has examined. This finding indicates that neuroinflammatory mechanisms and/or reactive oxygen species (ROS) should be investigated as specific targets for therapeutic intervention in GWI. She also observed reduced neurogenesis, reduced synaptophysin (a marker of pre-synaptic vesicles) as well as evidence for mitochondria and calcium signaling dysfunction. In addition, she has shown that compared to controls, exposed mice showed a significant decrease in a series of plasma-based pro-inflammatory cytokines.

This data suggest an anergic immune status in Gulf War agent-exposed mice compared to placebo-treated controls. Dr. Ait-Ghezala is continuing to scrutinize the altered neurobehavioral and neuropathological mechanisms in this animal model in the hope that it will help identify potential targets for therapeutic interventions to treat GWI.

Publications

Exposure to an organophosphate pesticide, individually or in combination with other Gulf War agents, impairs synaptic integrity and neuronal differentiation, and is accompanied by subtle microvascular injury in a mouse model of Gulf War agent exposure. [http://www.ncbi.nlm.nih.gov/pubmed/24118348] Ojo JO, Abdullah L, Evans J, Reed JM, Montague H, Mullan MJ, Crawford FC. Neuropathology. 2014 Apr;34(2):109-27. doi: 10.1111/neup.12061. Epub 2013 Sep 30.

Chronic elevation of phosphocholine containing lipids in mice exposed to Gulf War agents pyridostigmine bromide and permethrin. [http://www.ncbi.nlm.nih.gov/pubmed/24140745] Abdullah L, Evans JE, Montague H, Reed JM, Moser A, Crynen G, Gonzalez A, Zakirova Z, Ross I, Mullan C, Mullan M, Ait-Ghezala G, Crawford F. Neurotoxicol Teratol. 2013 Nov-Dec;40:74-84.

Lipidomic Profiling of Phosphocholine Containing Brain Lipids in Mice with Sensorimotor Deficits and Anxiety-Like Features After Exposure to Gulf War Agents. [http://www.ncbi.nlm.nih.gov/pubmed/22798222] Abdullah L, Evans JE, Bishop A, Reed JM, Crynen G, Phillips J, Pelot R, Mullan MA, Ferro A, Mullan CM, Mullan MJ, Ait-Ghezala G, Crawford FC. Neuromolecular Med. 2012

Proteomic CNS Profile of Delayed Cognitive Impairment in Mice Exposed to Gulf War Agents. [http://www.ncbi.nlm.nih.gov/pubmed/21986894]Abdullah L, Crynen G, Reed J, Bishop A, Phillips J, Ferguson S, Mouzon B, Mullan M, Mathura V, Mullan M, Ait-Ghezala G, Crawford F. Neuromolecular Med. 2011 13(4):275-88

Gulf War agents trigger discrete transcriptional changes in cultured human neurons. [http://www.tandfonline.com/doi/full/10.1080/02772241003611946] Kayihan GC, Wood M, Mouzon B, Ferguson S, Mullan M and Crawford F. Journal of Toxicology and Environmental Chem. 2010 92(9) 1783-1799.

Proteomic Analysis of Human Neuronal Cells Treated with the Gulf WarAgent Pyridostigmine Bromide. [http://omicsonline.org/ArchiveJPB/2009/October/03/JPB2.439.php?aid=1406] Abdullah L, Reed J, Kayihan G, Mathura V, Mouzon B, et al. (2009) J Proteomics Bioinform 2: 439-444.doi:10.4172/jpb.1000103.

Media and Presentations

Open House for Veterans and Military in honor of Veterans Day
Friday, November 7, 2014 10AM

Gulf War Illiness at Roskamp Institute

Our Gulf War Illness research was profiled in a Bradenton Herald Living section article.
Zuchra Zakirova

RI scientists highlight their Gulf War Illness program focus.
Gulf War Illness research

RI’s Gulf War Illness research was a featured story on ABC 7 News
Roskamp Institute NPR Gulf War Illness Article National Public Radio (NPR) Roskamp Institute Gulf War Illness Program Article

Treatment with Anatabine ameliorates cognitive impairment and neuropathological deficits in a mouse model of Gulf War Illness. Ait-Ghezala, Zakirova Z, Mouzon B, Crawford F, Mullan M, Mathura V. (will be) presented at: Annual Meeting Society for Neuroscience 2014

Proteomic profiling of the CNS in a mouse model of pyridostigmine bromide and permethrin exposure reveals mitochondrial dysfunction. Zakirova Z, Tweed M, Reed J, Crynen G, Hart A, Prashanthi V, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. (will be) presented at: Annual Meeting Society for Neuroscience 2014.

Assessment of neurobehavioral and neuropathological outcomes in a mouse model

Assessment of neurobehavioral and neuropathological outcomes in a mouse model of Gulf War Illness at early and late time points post exposure. Zakirova Z, Tweed M, Crynen G, Reed J, Abdullah L, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. BioFlorida Chapter meeting at New College of Florida 2014 and Tampa VA Annual Research Day 2014.

The relationship between cognitive impairment and inflammatory responses following exposure to Gulf War agents

The relationship between cognitive impairment and inflammatory responses following exposure to Gulf War agents at early and late time points using a mouse model of GWI. Zakirova Z, Tweed M, Crynen G, Reed J, Abdullah L, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. Annual Meeting Society for Neuroscience 2013.

Mouse model of Gulf War Illness reveals neurobehavioral and neuropathological correlates in Young and Aged Cohorts

Mouse model of Gulf War Illness reveals neurobehavioral and neuropathological correlates in Young and Aged Cohorts. Zakirova Z, Tweed M, Crynen G, Reed J, Abdullah L, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. Tampa VA Annual Research Day 2013.

Analysis of Neurobehavioral, Neuropathological and Proteomic Data from a mouse model of Gulf War Illness

Analysis of Neurobehavioral, Neuropathological and Proteomic Data from a mouse model of Gulf War Illness. Zakirova Z, Tweed M, Crynen G, Reed J, Hart A, Dufresne GP, Nissanka N, Bishop A, Abdullah L, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. Integrative. Annual Meeting Society for Neuroscience 2012.

Proteomic Profiling of Associated Neurobehavior and Cognitive Impairment in a Mouse Model of GWI.

Proteomic Profiling of Associated Neurobehavior and Cognitive Impairment in a Mouse Model of GWI. Zakirova Z, Nissanka N, Reed J, Bishop A, Abdullah L, Crynen G, Crawford F, Mullan M, Mathura V, Ait-Ghezala G. Tampa VA Annual Research Day 2012.

GWI Research Project Funding:

Listed below are GWI grants and contracts awarded to the Roskamp Institute (Years/Research Title/Grantor):

2014-2017
Restoring brain lipid homeostasis as a therapeutic avenue for treating Gulf War Illness. (Abdullah/Crawford)
Congressionally Directed Medical Research Program

2013-2017
Identification of plasma biomarkers of Gulf War Illness using omicâ€ technology. (Crawford)
VA Merit award

2011-2014
Proteomic immune profiling for the therapeutic modulation of cognitive impairment in a novel Gulf War Illness mouse model. (Ait-Ghezala)
Congressionally Directed Medical Research Program

2009-2012
Identification of biological pathways implicated in hippocampal dysfunction and cognitive impairment in Gulf War Illness. (Crawford)
Congressionally Directed Medical Research Program

2006-2009
Proteomic analysis of cellular response to biological warfare agents. (Crawford/Mullan);
VA Merit Award

The GWI Research Program is also supported by the Roskamp Foundation.