Time- and dose-dependent analysis of gene expression using microarrays in sulfur mustard-exposed mice

Abstract

The chemical warfare agent sulfur mustard (SM) produces blister formation with a severe inflammatory reaction in skin of exposed individuals. The development of efficacious countermeasures against SM vesication requires an understanding of the cellular and molecular mechanism of SM-induced tissue injury. This study examined SM-induced alterations in gene expression using Atlas Mouse 5K DNA microarrays (5002 genes) to identify transcriptional events associated with SM skin injury. Mice (N=3) were exposed topically to SM (0.04, 0.08, and 0.16 mg; 48.8, 97.5, and 195 mM) on the inner surface of the right ear and skin tissues were harvested at 1.5, 3, 6, and 12 h. Genes were selected based on the three mice in the same dose group demonstrating a > or =2-fold increase or decrease in gene expression for the SM-exposed tissue when compared to the dichloromethane vehicle control ear at all three doses and four time points. At the 0.04 mg SM dose, the genes observed were primarily involved in inflammation, apoptosis, and cell cycle regulation. Exposure to 0.08 mg SM increased the expression of genes related to inflammation and cell cycle regulation. Exposure to 0.16 mg SM led to a total of six genes that were changed at all observed time periods; however, these genes do not appear to be directly influential in biological mechanisms such as inflammation, apoptosis, and cell cycle regulation as was observed at the lower SM doses of 0.04 and 0.08 mg. These functional categories have been observed in previous studies utilizing both in vivo and in vitro model systems of SM-induced dermal injury, suggesting that molecular mechanisms associated with inflammation, apoptosis, and cell cycle regulation may be appropriate targets for developing prophylactic/therapeutic treatments for SM skin injury.

Time course of skin features and inflammatory biomarkers after liquid sulfur mustard exposure in SKH-1 hairless mice

“…This skin inflammation in response to SM exposure is controlled by an important regulation of a network of inflammatory mediators to recruit and activate inflammatory cells to the site of injury. Accordingly, several animal studies have documented an increased expression of interleukin (IL)-1a, IL-1b, IL-6, IL-8, tumor necrosis factor (TNF)-a, macrophage inflammatory proteins (MIP)-1a, MIP2a, MIP-1aR, granulocyte monocyte-colony stimulating factor (GM-CSF), cyclooxygenase-2 (COX2) and inducible nitric oxide synthase following vapor or liquid SM exposure (Nyska et al, 2001;Price et al, 2009;Ricketts et al, 2000;Sabourin et al, 2000Sabourin et al, , 2004Shakarjian et al, 2010;Vallet et al, 2012;Wormser et al, 2004Wormser et al, , 2005.…”

“…Several animal models including weanling pigs (Sabourin et al, 2002;Smith et al, 1996Smith et al, , 1997b, hairless guinea pig (Benson et al, 2011;Dachir et al, 2010Dachir et al, , 2012Smith et al, 1997a), rabbit (Dannenberg et al, 1985) or various mouse species (Lomash et al, 2011(Lomash et al, , 2013Powers et al, 2000;Ricketts et al, 2000;Sabourin et al, 2004;Wormser et al, 2005) have been used to investigate SMinduced skin toxicity. During the last years, the hairless mouse model stood out as the model of choice for studying the skin pathogenesis of SM Dorandeu et al, 2011;Joseph et al, 2011;Vallet et al, 2012) or its analogues the chloroethyl ethyl sulfide (CEES) (Jain et al, 2011a,b,b;Tewari-Singh et al, 2009) or the nitrogen mustard Jain et al, 2014;Tewari-Singh et al, 2013.…”

Microarray Analysis of Mouse Ear Tissue Exposed to Bis-(2-chloroethyl) Sulfide: Gene Expression Profiles Correlate with Treatment Efficacy and An Established Clinical Endpoint

“…The systems studied include mouse ear (Sabourin et al, 2000(Sabourin et al, , 2004aRogers et al, 2004), pig skin (Sabourin et al, 2002), cultured human keratinocytes Fig. 6.…”

“…Up-regulation of cytokines and chemokines after SM exposure has been well characterized in a number of recent studies (Arroyo et al, 1999;Lardot et al, 1999;Sabourin et al, 2000Sabourin et al, , 2002Sabourin et al, , 2003Sabourin et al, , 2004a. Related to this, a number of cell signaling pathways have been identified that are activated or altered after SM exposure.…”

“…The mechanism of toxicity of SM is not well characterized, and previous studies have used DNA microarray technology to gain greater insight into the molecular pathways perturbed by SM exposure (Rogers et al, 2004;Sabourin et al, 2004a). Based on these studies, we hypothesized that gene expression profiling of mouse ear skin exposed to SM alone or pretreated with one of these treatment compounds would provide important insight into the mechanism of cutaneous toxicity of SM and might identify genes and biological pathways involved in the response to SM exposure.…”