Differential gene expression profiling of mouse skin after sulfur mustard exposure: Extended time response and inhibitor effect

Environmental and Occupational Health Sciences Institute (EOHSI), a Joint Institute of UMDNJ-RW Johnson Medical School and Rutgers University, 170 Frelinghuysen Road, Piscataway, NJ 08854, USA.
Toxicology and Applied Pharmacology (Impact Factor: 3.71). 11/2008; 234(2):156-65. DOI: 10.1016/j.taap.2008.09.020
Source: PubMed


Sulfur mustard (HD, SM), is a chemical warfare agent that within hours causes extensive blistering at the dermal-epidermal junction of skin. To better understand the progression of SM-induced blistering, gene expression profiling for mouse skin was performed after a single high dose of SM exposure. Punch biopsies of mouse ears were collected at both early and late time periods following SM exposure (previous studies only considered early time periods). The biopsies were examined for pathological disturbances and the samples further assayed for gene expression profiling using the Affymetrix microarray analysis system. Principal component analysis and hierarchical cluster analysis of the differently expressed genes, performed with ArrayTrack showed clear separation of the various groups. Pathway analysis employing the KEGG library and Ingenuity Pathway Analysis (IPA) indicated that cytokine-cytokine receptor interaction, cell adhesion molecules (CAMs), and hematopoietic cell lineage are common pathways affected at different time points. Gene ontology analysis identified the most significantly altered biological processes as the immune response, inflammatory response, and chemotaxis; these findings are consistent with other reported results for shorter time periods. Selected genes were chosen for RT-PCR verification and showed correlations in the general trends for the microarrays. Interleukin 1 beta was checked for biological analysis to confirm the presence of protein correlated to the corresponding microarray data. The impact of a matrix metalloproteinase inhibitor, MMP-2/MMP-9 inhibitor I, against SM exposure was assessed. These results can help in understanding the molecular mechanism of SM-induced blistering, as well as to test the efficacy of different inhibitors.

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    • "P.A. Everley, J.F. Dillman, 3rd / Toxicology Letters 198 (2010) 297–303 Table 1 List of systems biology-related genomics and proteomics studies used in CWA research. CWA Genomics/proteomics Study GB Genomics Pachiappan et al. (2009) Damodaran et al. (2006a) Damodaran et al. (2006b) GD Genomics Dillman et al. (2009) VX Genomics Blanton et al. (2004) HD Genomics Price et al. (2009) Gerecke et al. (2009) Rogers et al. (2008) Yu et al. (2006) Dillman et al. (2006) Dillman et al. (2005) Rogers et al. (2004) Shahin et al. (2001) Lakshmana Rao et al. (1999) Meier and Millard (1998) Proteomics Everley and Dillman (2010) Mehrani et al. (2009) Mol et al. (2008) An et al. (2006) Dillman et al. (2003) Dillman and Schlager (2003) Phosgene Genomics Sciuto et al. (2005) MEDLINE, ToxFile, Biosis Previews, EMBASE and CA SEARCH databases were searched for articles containing " genomic " or " proteomic " terms using the following keywords: sarin (GB), soman (GD), VX, VR, sulfur mustard (HD), chlorine, cyanide, cyclosarin (GF), and phosgene. Articles returned from the database search were filtered for CWA relevance. "
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    ABSTRACT: Medical research on the effects of chemical warfare agents (CWAs) has been ongoing for nearly 100 years, yet these agents continue to pose a serious threat to deployed military forces and civilian populations. CWAs are extremely toxic, relatively inexpensive, and easy to produce, making them a legitimate weapon of choice for terrorist organizations. While the mechanisms of action for many CWAs have been known for years, questions about their molecular effects following acute and chronic exposure remain largely unanswered. Global approaches that can pinpoint which cellular pathways are altered in response to CWAs and characterize long-term toxicity have not been widely used. Fortunately, innovations in genomics and proteomics technologies now allow for thousands of genes and proteins to be identified and subsequently quantified in a single experiment. Advanced bioinformatics software can also help decipher large-scale changes observed, leading to mapping of signaling pathways, functional characterization, and identification of potential therapeutic targets. Here we present an overview of how genomics and proteomics technologies have been applied to CWA research and also provide a series of questions focused on how these techniques could further our understanding of CWA toxicity.
    Toxicology Letters 10/2010; 198(3):297-303. DOI:10.1016/j.toxlet.2010.08.003 · 3.26 Impact Factor
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    • "In a recent study, migration of human epidermal keratinocytes, in in vitro scratch wounds, was inhibited by LPS and the inhibition could be blocked partially by anti-TLR4 (75%) and anti-TLR2 (40%) [26]. The emerging roles of TLRs in wound healing has received little experimental attention to date except in chemical or thermal-induced burn injuries [27-31]. For example, TLR4 plays a critical role in microvascular leakage and leukocyte adhesion under the inflammatory conditions associated with nonseptic thermal injury [27]. "
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    ABSTRACT: When compared to skin, oral mucosal wounds heal rapidly and with reduced scar formation. Recent studies suggest that intrinsic differences in inflammation, growth factor production, levels of stem cells, and cellular proliferation capacity may underlie the exceptional healing that occurs in oral mucosa. The current study was designed to compare the transcriptomes of oral mucosal and skin wounds in order to identify critical differences in the healing response at these two sites using an unbiased approach. Using microarray analysis, we explored the differences in gene expression in skin and oral mucosal wound healing in a murine model of paired equivalent sized wounds. Samples were examined from days 0 to 10 and spanned all stages of the wound healing process. Using unwounded matched tissue as a control, filtering identified 1,479 probe sets in skin wounds yet only 502 probe sets in mucosal wounds that were significantly differentially expressed over time. Clusters of genes that showed similar patterns of expression were also identified in each wound type. Analysis of functionally related gene expression demonstrated dramatically different reactions to injury between skin and mucosal wounds. To explore whether site-specific differences might be derived from intrinsic differences in cellular responses at each site, we compared the response of isolated epithelial cells from skin and oral mucosa to a defined in vitro stimulus. When cytokine levels were measured, epithelial cells from skin produced significantly higher amounts of proinflammatory cytokines than cells from oral mucosa. The results provide the first detailed molecular profile of the site-specific differences in the genetic response to injury in mucosa and skin, and suggest the divergent reactions to injury may derive from intrinsic differences in the cellular responses at each site.
    BMC Genomics 08/2010; 11(1):471. DOI:10.1186/1471-2164-11-471 · 3.99 Impact Factor
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    • "A correlation has been reported between the changes in several of these genes and the ability of antivesicants to reduce SMinduced ear edema. Significant changes in gene expression have also been reported up to 7 days following exposure of mice to SM (Gerecke et al., 2009). Gene ontology analysis suggests that the most significantly altered biological processes are immune and inflammatory responses and that the specific genes expressed differ as a function of time after exposure. "
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    ABSTRACT: Sulfur mustard (SM), a chemical weapon first employed during WorldWar I, targets the skin, eyes, and lung. It remains a significant military and civilian threat. The characteristic response of human skin to SM involves erythema of delayed onset, followed by edema with inflammatory cell infiltration, the appearance of large blisters in the affected area, and a prolonged healing period. Several in vivo and in vitro models have been established to understand the pathology and investigate the mechanism of action of this vesicating agent in the skin. SM is a bifunctional alkylating agent which reacts with many targets including lipids, proteins, and DNA, forming both intra-and intermolecular cross-links. Despite the relatively nonselective chemical reactivity of this agent, basal keratinocytes are more sensitive, and blistering involves detachment of these cells from their basement membrane adherence zones. The sequence and manner in which these cells die and detach is still unresolved. Much has been discovered over the past two decades with respect to the mechanisms of SM-induced cytotoxicity and the intracellular and extracellular targets of this vesicant. In this review, the effects of SM exposure on the skin are described, as well as potential mechanisms mediating its actions. Successful therapy for SM poisoning will depend on following new mechanistic leads to develop drugs that target one or more of its sites of action. © The Author 2009. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: [email protected] /* */
    Toxicological Sciences 10/2009; 114(1):5-19. DOI:10.1093/toxsci/kfp253 · 3.85 Impact Factor
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