The Experts below are selected from a list of 360 Experts worldwide ranked by ideXlab platform
Debra L Laskin - One of the best experts on this subject based on the ideXlab platform.
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nutraceuticals as potential therapeutics for Vesicant induced pulmonary fibrosis
Annals of the New York Academy of Sciences, 2020Co-Authors: Rita Businaro, Elisa Maggi, Federica Armeli, Alexa Murray, Debra L LaskinAbstract:Exposure to Vesicants, including sulfur mustard and nitrogen mustard, causes damage to the epithelia of the respiratory tract and the lung. With time, this progresses to chronic disease, most notably, pulmonary fibrosis. The pathogenic process involves persistent inflammation and the release of cytotoxic oxidants, cytokines, chemokines, and profibrotic growth factors, which leads to the collapse of lung architecture, with fibrotic involution of the lung parenchyma. At present, there are no effective treatments available to combat this pathological process. Recently, much interest has focused on nutraceuticals, substances derived from plants, herbs, and fruits, that exert pleiotropic effects on inflammatory cells and parenchymal cells that may be useful in reducing fibrogenesis. Some promising results have been obtained with nutraceuticals in experimental animal models of inflammation-driven fibrosis. This review summarizes the current knowledge on the putative preventive/therapeutic efficacy of nutraceuticals in progressive pulmonary fibrosis, with a focus on their activity against inflammatory reactions and profibrotic cell differentiation.
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Sulfur Mustard Analog Mechlorethamine (Bis(2-chloroethyl)methylamine) Modulates Cell Cycle Progression via the DNA Damage Response in Human Lung Epithelial A549 Cells
2019Co-Authors: Yi-hua Jan, Diane E Heck, Debra L Laskin, Jeffrey D LaskinAbstract:Nitrogen mustard, mechlorethamine (bis(2-chloroethyl)methylamine; HN2), and sulfur mustard are potent Vesicants that modify and disrupt cellular macromolecules including DNA leading to cytotoxicity and tissue injury. In many cell types, HN2 upregulates DNA damage signaling pathways including ataxia telangiectasia mutated (ATM), ataxia telangiectasia mutated- and Rad3-related (ATR) as well as DNA-dependent protein kinase (DNA-PK). In the present studies, we investigated crosstalk between the HN2-induced DNA damage response and cell cycle progression using human A549 lung epithelial cells. HN2 (1–20 μM; 24 h) caused a concentration-dependent arrest of cells in the S and G2/M phases of the cell cycle. This was associated with inhibition of DNA synthesis, as measured by incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into S phase cells. Cell cycle arrest was correlated with activation of DNA damage and cell cycle checkpoint signaling. Thus, HN2 treatment resulted in time- and concentration-dependent increases in expression of phosphorylated ATM (Ser1981), Chk2 (Thr68), H2AX (Ser139), and p53 (Ser15). Activation of DNA damage signaling was most pronounced in S-phase cells followed by G2/M-phase cells. HN2-induced cell cycle arrest was suppressed by the ATM and DNA-PK inhibitors, KU55933 and NU7441, respectively, and to a lesser extent by VE821, an ATR inhibitor. This was correlated with abrogation of DNA damage checkpoints signaling. These data indicate that activation of ATM, ATR, and DNA-PK signaling pathways by HN2 are important in the mechanism of Vesicant-induced cell cycle arrest and cytotoxicity. Drugs that inhibit activation of DNA damage signaling may be effective countermeasures for Vesicant-induced tissue injury
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mustard Vesicant induced lung injury advances in therapy
Toxicology and Applied Pharmacology, 2016Co-Authors: Barry Weinberger, Diane E Heck, Jeffrey D Laskin, Vasanthi R Sunil, Rama Malaviya, Alessandro Venosa, Debra L LaskinAbstract:Most mortality and morbidity following exposure to Vesicants such as sulfur mustard is due to pulmonary toxicity. Acute injury is characterized by epithelial detachment and necrosis in the pharynx, trachea and bronchioles, while long-term consequences include fibrosis and, in some instances, cancer. Current therapies to treat mustard poisoning are primarily palliative and do not target underlying pathophysiologic mechanisms. New knowledge about Vesicant-induced pulmonary disease pathogenesis has led to the identification of potentially efficacious strategies to reduce injury by targeting inflammatory cells and mediators including reactive oxygen and nitrogen species, proteases and proinflammatory/cytotoxic cytokines. Therapeutics under investigation include corticosteroids, N-acetyl cysteine, which has both mucolytic and antioxidant properties, inducible nitric oxide synthase inhibitors, liposomes containing superoxide dismutase, catalase, and/or tocopherols, protease inhibitors, and cytokine antagonists such as anti-tumor necrosis factor (TNF)-α antibody and pentoxifylline. Antifibrotic and fibrinolytic treatments may also prove beneficial in ameliorating airway obstruction and lung remodeling. More speculative approaches include inhibitors of transient receptor potential channels, which regulate pulmonary epithelial cell membrane permeability, non-coding RNAs and mesenchymal stem cells. As mustards represent high priority chemical threat agents, identification of effective therapeutics for mitigating toxicity is highly significant.
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increased expression of the endocannabinoid system in mouse skin following exposure to sulfur mustard and nitrogen mustard mechlorethamine
The FASEB Journal, 2016Co-Authors: Diane E Heck, Donald R Gerecke, Debra L Laskin, Robert P Casillas, Irene Wohlman, Gabriella M Composto, Ned D Heindel, Laurie B Joseph, Jeffrey D LaskinAbstract:Vesicants including sulfur mustard (SM, bis(2-chloroethyl) sulfide) and nitrogen mustard (NM, bis(2-chloroethyl)methylamine) are highly reactive bifunctional alkylating agents that target the skin....
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role of reactive nitrogen species generated via inducible nitric oxide synthase in Vesicant induced lung injury inflammation and altered lung functioning
Toxicology and Applied Pharmacology, 2012Co-Authors: Vasanthi R Sunil, Jeffrey D Laskin, Jianliang Shen, Kinal Patelvayas, Andrew J Gow, Debra L LaskinAbstract:Pulmonary toxicity induced by Vesicants is associated with oxidative stress. In the present studies we analyzed the role of reactive nitrogen species (RNS) generated via inducible nitric oxide synthase (iNOS) in lung injury and inflammation induced by Vesicants using 2-chloroethyl ethyl sulfide (CEES) as a model. C57Bl/6 (WT) and iNOS−/− mice were sacrificed 3 d or 14 d following intratracheal administration of CEES (6 mg/kg) or control. CEES intoxication resulted in transient (3 d) increases in bronchoalveolar lavage (BAL) cell and protein content in WT, but not iNOS−/− mice. This correlated with expression of Ym1, a marker of oxidative stress in alveolar macrophages and epithelial cells. In contrast, in iNOS−/− mice, Ym1 was only observed 14 d post exposure in enlarged alveolar macrophages, suggesting that they are alternatively activated. This is supported by findings that lung tumor necrosis factor and lipocalin Lcn2 expression, mediators involved in tissue repair were also upregulated at this time in iNOS−/− mice. Conversely, CEES-induced increases in the proinflammatory genes, monocyte chemotactic protein-1 and cyclooxygenase-2, were abrogated in iNOS−/− mice. In WT mice, CEES treatment also resulted in increases in total lung resistance and decreases in compliance in response to methacholine, effects blunted by loss of iNOS. These data demonstrate that RNS, generated via iNOS play a role in the pathogenic responses to CEES, augmenting oxidative stress and inflammation and suppressing tissue repair. Elucidating inflammatory mechanisms mediating Vesicant-induced lung injury is key to the development of therapeutics to treat mustard poisoning.
Jeffrey D Laskin - One of the best experts on this subject based on the ideXlab platform.
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Sulfur Mustard Analog Mechlorethamine (Bis(2-chloroethyl)methylamine) Modulates Cell Cycle Progression via the DNA Damage Response in Human Lung Epithelial A549 Cells
2019Co-Authors: Yi-hua Jan, Diane E Heck, Debra L Laskin, Jeffrey D LaskinAbstract:Nitrogen mustard, mechlorethamine (bis(2-chloroethyl)methylamine; HN2), and sulfur mustard are potent Vesicants that modify and disrupt cellular macromolecules including DNA leading to cytotoxicity and tissue injury. In many cell types, HN2 upregulates DNA damage signaling pathways including ataxia telangiectasia mutated (ATM), ataxia telangiectasia mutated- and Rad3-related (ATR) as well as DNA-dependent protein kinase (DNA-PK). In the present studies, we investigated crosstalk between the HN2-induced DNA damage response and cell cycle progression using human A549 lung epithelial cells. HN2 (1–20 μM; 24 h) caused a concentration-dependent arrest of cells in the S and G2/M phases of the cell cycle. This was associated with inhibition of DNA synthesis, as measured by incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into S phase cells. Cell cycle arrest was correlated with activation of DNA damage and cell cycle checkpoint signaling. Thus, HN2 treatment resulted in time- and concentration-dependent increases in expression of phosphorylated ATM (Ser1981), Chk2 (Thr68), H2AX (Ser139), and p53 (Ser15). Activation of DNA damage signaling was most pronounced in S-phase cells followed by G2/M-phase cells. HN2-induced cell cycle arrest was suppressed by the ATM and DNA-PK inhibitors, KU55933 and NU7441, respectively, and to a lesser extent by VE821, an ATR inhibitor. This was correlated with abrogation of DNA damage checkpoints signaling. These data indicate that activation of ATM, ATR, and DNA-PK signaling pathways by HN2 are important in the mechanism of Vesicant-induced cell cycle arrest and cytotoxicity. Drugs that inhibit activation of DNA damage signaling may be effective countermeasures for Vesicant-induced tissue injury
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mustard Vesicant induced lung injury advances in therapy
Toxicology and Applied Pharmacology, 2016Co-Authors: Barry Weinberger, Diane E Heck, Jeffrey D Laskin, Vasanthi R Sunil, Rama Malaviya, Alessandro Venosa, Debra L LaskinAbstract:Most mortality and morbidity following exposure to Vesicants such as sulfur mustard is due to pulmonary toxicity. Acute injury is characterized by epithelial detachment and necrosis in the pharynx, trachea and bronchioles, while long-term consequences include fibrosis and, in some instances, cancer. Current therapies to treat mustard poisoning are primarily palliative and do not target underlying pathophysiologic mechanisms. New knowledge about Vesicant-induced pulmonary disease pathogenesis has led to the identification of potentially efficacious strategies to reduce injury by targeting inflammatory cells and mediators including reactive oxygen and nitrogen species, proteases and proinflammatory/cytotoxic cytokines. Therapeutics under investigation include corticosteroids, N-acetyl cysteine, which has both mucolytic and antioxidant properties, inducible nitric oxide synthase inhibitors, liposomes containing superoxide dismutase, catalase, and/or tocopherols, protease inhibitors, and cytokine antagonists such as anti-tumor necrosis factor (TNF)-α antibody and pentoxifylline. Antifibrotic and fibrinolytic treatments may also prove beneficial in ameliorating airway obstruction and lung remodeling. More speculative approaches include inhibitors of transient receptor potential channels, which regulate pulmonary epithelial cell membrane permeability, non-coding RNAs and mesenchymal stem cells. As mustards represent high priority chemical threat agents, identification of effective therapeutics for mitigating toxicity is highly significant.
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increased expression of the endocannabinoid system in mouse skin following exposure to sulfur mustard and nitrogen mustard mechlorethamine
The FASEB Journal, 2016Co-Authors: Diane E Heck, Donald R Gerecke, Debra L Laskin, Robert P Casillas, Irene Wohlman, Gabriella M Composto, Ned D Heindel, Laurie B Joseph, Jeffrey D LaskinAbstract:Vesicants including sulfur mustard (SM, bis(2-chloroethyl) sulfide) and nitrogen mustard (NM, bis(2-chloroethyl)methylamine) are highly reactive bifunctional alkylating agents that target the skin....
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sulfur mustard induces an endoplasmic reticulum stress response in the mouse ear Vesicant model
Toxicology and Applied Pharmacology, 2013Co-Authors: Yokechen Chang, Marion K Gordon, Jeffrey D Laskin, Robert P Casillas, James D Wang, Kathy K H Svoboda, Donald R GereckeAbstract:The endoplasmic reticulum (ER) stress response is a cell survival pathway upregulated when cells are under severe stress. Severely damaged mouse ear skin exposed to the Vesicant, sulfur mustard (bis-2-chloroethyl sulfide, SM), resulted in increased expression of ER chaperone proteins that accompany misfolded and incorrectly made proteins targeted for degradation. Time course studies with SM using the mouse ear Vesicant model (MEVM) showed progressive histopathologic changes including edema, separation of the epidermis from the dermis, persistent inflammation, upregulation of laminin γ2 (one of the chains of laminin-332, a heterotrimeric skin glycoprotein required for wound repair), and delayed wound healing from 24 h to 168 h post exposure. This was associated with time related increased expression of the cell survival ER stress marker, GRP78/BiP, and the ER stress apoptosis marker, GADD153/CHOP, suggesting simultaneous activation of both cell survival and non-mitochondrial apoptosis pathways. Dual immunofluorescence labeling of a keratinocyte migration promoting protein, laminin γ2 and GRP78/BIP, showed colocalization of the two molecules 72 h post exposure indicating that the laminin γ2 was misfolded after SM exposure and trapped within the ER. Taken together, these data show that ER stress is induced in mouse skin within 24 h of Vesicant exposure in a defensive response to promote cell survival; however, it appears that this response is rapidly overwhelmed by the apoptotic pathway as a consequence of severe SM-induced injury.
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role of reactive nitrogen species generated via inducible nitric oxide synthase in Vesicant induced lung injury inflammation and altered lung functioning
Toxicology and Applied Pharmacology, 2012Co-Authors: Vasanthi R Sunil, Jeffrey D Laskin, Jianliang Shen, Kinal Patelvayas, Andrew J Gow, Debra L LaskinAbstract:Pulmonary toxicity induced by Vesicants is associated with oxidative stress. In the present studies we analyzed the role of reactive nitrogen species (RNS) generated via inducible nitric oxide synthase (iNOS) in lung injury and inflammation induced by Vesicants using 2-chloroethyl ethyl sulfide (CEES) as a model. C57Bl/6 (WT) and iNOS−/− mice were sacrificed 3 d or 14 d following intratracheal administration of CEES (6 mg/kg) or control. CEES intoxication resulted in transient (3 d) increases in bronchoalveolar lavage (BAL) cell and protein content in WT, but not iNOS−/− mice. This correlated with expression of Ym1, a marker of oxidative stress in alveolar macrophages and epithelial cells. In contrast, in iNOS−/− mice, Ym1 was only observed 14 d post exposure in enlarged alveolar macrophages, suggesting that they are alternatively activated. This is supported by findings that lung tumor necrosis factor and lipocalin Lcn2 expression, mediators involved in tissue repair were also upregulated at this time in iNOS−/− mice. Conversely, CEES-induced increases in the proinflammatory genes, monocyte chemotactic protein-1 and cyclooxygenase-2, were abrogated in iNOS−/− mice. In WT mice, CEES treatment also resulted in increases in total lung resistance and decreases in compliance in response to methacholine, effects blunted by loss of iNOS. These data demonstrate that RNS, generated via iNOS play a role in the pathogenic responses to CEES, augmenting oxidative stress and inflammation and suppressing tissue repair. Elucidating inflammatory mechanisms mediating Vesicant-induced lung injury is key to the development of therapeutics to treat mustard poisoning.
Diane E Heck - One of the best experts on this subject based on the ideXlab platform.
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Sulfur Mustard Analog Mechlorethamine (Bis(2-chloroethyl)methylamine) Modulates Cell Cycle Progression via the DNA Damage Response in Human Lung Epithelial A549 Cells
2019Co-Authors: Yi-hua Jan, Diane E Heck, Debra L Laskin, Jeffrey D LaskinAbstract:Nitrogen mustard, mechlorethamine (bis(2-chloroethyl)methylamine; HN2), and sulfur mustard are potent Vesicants that modify and disrupt cellular macromolecules including DNA leading to cytotoxicity and tissue injury. In many cell types, HN2 upregulates DNA damage signaling pathways including ataxia telangiectasia mutated (ATM), ataxia telangiectasia mutated- and Rad3-related (ATR) as well as DNA-dependent protein kinase (DNA-PK). In the present studies, we investigated crosstalk between the HN2-induced DNA damage response and cell cycle progression using human A549 lung epithelial cells. HN2 (1–20 μM; 24 h) caused a concentration-dependent arrest of cells in the S and G2/M phases of the cell cycle. This was associated with inhibition of DNA synthesis, as measured by incorporation of 5-ethynyl-2′-deoxyuridine (EdU) into S phase cells. Cell cycle arrest was correlated with activation of DNA damage and cell cycle checkpoint signaling. Thus, HN2 treatment resulted in time- and concentration-dependent increases in expression of phosphorylated ATM (Ser1981), Chk2 (Thr68), H2AX (Ser139), and p53 (Ser15). Activation of DNA damage signaling was most pronounced in S-phase cells followed by G2/M-phase cells. HN2-induced cell cycle arrest was suppressed by the ATM and DNA-PK inhibitors, KU55933 and NU7441, respectively, and to a lesser extent by VE821, an ATR inhibitor. This was correlated with abrogation of DNA damage checkpoints signaling. These data indicate that activation of ATM, ATR, and DNA-PK signaling pathways by HN2 are important in the mechanism of Vesicant-induced cell cycle arrest and cytotoxicity. Drugs that inhibit activation of DNA damage signaling may be effective countermeasures for Vesicant-induced tissue injury
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mustard Vesicant induced lung injury advances in therapy
Toxicology and Applied Pharmacology, 2016Co-Authors: Barry Weinberger, Diane E Heck, Jeffrey D Laskin, Vasanthi R Sunil, Rama Malaviya, Alessandro Venosa, Debra L LaskinAbstract:Most mortality and morbidity following exposure to Vesicants such as sulfur mustard is due to pulmonary toxicity. Acute injury is characterized by epithelial detachment and necrosis in the pharynx, trachea and bronchioles, while long-term consequences include fibrosis and, in some instances, cancer. Current therapies to treat mustard poisoning are primarily palliative and do not target underlying pathophysiologic mechanisms. New knowledge about Vesicant-induced pulmonary disease pathogenesis has led to the identification of potentially efficacious strategies to reduce injury by targeting inflammatory cells and mediators including reactive oxygen and nitrogen species, proteases and proinflammatory/cytotoxic cytokines. Therapeutics under investigation include corticosteroids, N-acetyl cysteine, which has both mucolytic and antioxidant properties, inducible nitric oxide synthase inhibitors, liposomes containing superoxide dismutase, catalase, and/or tocopherols, protease inhibitors, and cytokine antagonists such as anti-tumor necrosis factor (TNF)-α antibody and pentoxifylline. Antifibrotic and fibrinolytic treatments may also prove beneficial in ameliorating airway obstruction and lung remodeling. More speculative approaches include inhibitors of transient receptor potential channels, which regulate pulmonary epithelial cell membrane permeability, non-coding RNAs and mesenchymal stem cells. As mustards represent high priority chemical threat agents, identification of effective therapeutics for mitigating toxicity is highly significant.
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increased expression of the endocannabinoid system in mouse skin following exposure to sulfur mustard and nitrogen mustard mechlorethamine
The FASEB Journal, 2016Co-Authors: Diane E Heck, Donald R Gerecke, Debra L Laskin, Robert P Casillas, Irene Wohlman, Gabriella M Composto, Ned D Heindel, Laurie B Joseph, Jeffrey D LaskinAbstract:Vesicants including sulfur mustard (SM, bis(2-chloroethyl) sulfide) and nitrogen mustard (NM, bis(2-chloroethyl)methylamine) are highly reactive bifunctional alkylating agents that target the skin....
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regulation of hsp27 and hsp70 expression in human and mouse skin construct models by caveolae following exposure to the model sulfur mustard Vesicant 2 chloroethyl ethyl sulfide
Toxicology and Applied Pharmacology, 2011Co-Authors: Adrienne T Black, Donald R Gerecke, Diane E Heck, Debra L Laskin, Patrick J Hayden, Robert P Casillas, Patrick J Sinko, Jeffrey D LaskinAbstract:Dermal exposure to the Vesicant sulfur mustard causes marked inflammation and tissue damage. Basal keratinocytes appear to be a major target of sulfur mustard. In the present studies, mechanisms mediating skin toxicity were examined using a mouse skin construct model and a full-thickness human skin equivalent (EpiDerm-FT™). In both systems, administration of the model sulfur mustard Vesicant, 2-chloroethyl ethyl sulfide (CEES, 100-1000μM) at the air surface induced mRNA and protein expression of heat shock proteins 27 and 70 (Hsp27 and Hsp70). CEES treatment also resulted in increased expression of caveolin-1, the major structural component of caveolae. Immunohistochemistry revealed that Hsp27, Hsp70 and caveolin-1 were localized in basal and suprabasal layers of the epidermis. Caveolin-1 was also detected in fibroblasts in the dermal component of the full thickness human skin equivalent. Western blot analysis of caveolar membrane fractions isolated by sucrose density centrifugation demonstrated that Hsp27 and Hsp70 were localized in caveolae. Treatment of mouse keratinocytes with filipin III or methyl-β-cyclodextrin, which disrupt caveolar structure, markedly suppressed CEES-induced Hsp27 and Hsp70 mRNA and protein expression. CEES treatment is known to activate JNK and p38 MAP kinases; in mouse keratinocytes, inhibition of these enzymes suppressed CEES-induced expression of Hsp27 and Hsp70. These data suggest that MAP kinases regulate Hsp 27 and Hsp70; moreover, caveolae-mediated regulation of heat shock protein expression may be important in the pathophysiology of Vesicant-induced skin toxicity.
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expression of proliferative and inflammatory markers in a full thickness human skin equivalent following exposure to the model sulfur mustard Vesicant 2 chloroethyl ethyl sulfide
Toxicology and Applied Pharmacology, 2010Co-Authors: Adrienne T Black, Donald R Gerecke, Diane E Heck, Debra L Laskin, Patrick J Hayden, Robert P Casillas, Patrick J Sinko, Jeffrey D LaskinAbstract:a r t i c l e i n f o Article history: Sulfur mustard is a potent Vesicant that induces in! ammation, edema and blistering following dermal exposure. To assess molecular mechanisms mediating these responses, we analyzed the effects of the model sulfur mustard Vesicant, 2-chloroethyl ethyl sul" de, on EpiDerm-FT™, a commercially available full-thickness human skin equivalent. CEES (100-1000 ! M) caused a concentration-dependent increase in pyknotic nuclei and vacuolization in basal keratinocytes; at high concentrations (300-1000 ! M), CEES also disrupted keratin " lament architecture in the stratum corneum. This was associated with time-dependent increases in expression of proliferating cell nuclear antigen, a marker of cell proliferation, and poly(ADP-ribose) polymerase (PARP) and phosphorylated histone H2AX, markers of DNA damage. Concentration- and time- dependent increases in mRNA and protein expression of eicosanoid biosynthetic enzymes including COX-2, 5- lipoxygenase, microsomal PGE2 synthases, leukotriene (LT) A4 hydrolase and LTC4 synthase were observed in CEES-treated skin equivalents, as well as in antioxidant enzymes, glutathione S-transferases A1-2 (GSTA1-2), GSTA3 and GSTA4. These data demonstrate that CEES induces rapid cellular damage, cytotoxicity and in! ammation in full-thickness skin equivalents. These effects are similar to human responses to Vesicants in vivo and suggest that the full thickness skin equivalent is a useful in vitro model to characterize the biological effects of mustards and to develop potential therapeutics.
Vasanthi R Sunil - One of the best experts on this subject based on the ideXlab platform.
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mustard Vesicant induced lung injury advances in therapy
Toxicology and Applied Pharmacology, 2016Co-Authors: Barry Weinberger, Diane E Heck, Jeffrey D Laskin, Vasanthi R Sunil, Rama Malaviya, Alessandro Venosa, Debra L LaskinAbstract:Most mortality and morbidity following exposure to Vesicants such as sulfur mustard is due to pulmonary toxicity. Acute injury is characterized by epithelial detachment and necrosis in the pharynx, trachea and bronchioles, while long-term consequences include fibrosis and, in some instances, cancer. Current therapies to treat mustard poisoning are primarily palliative and do not target underlying pathophysiologic mechanisms. New knowledge about Vesicant-induced pulmonary disease pathogenesis has led to the identification of potentially efficacious strategies to reduce injury by targeting inflammatory cells and mediators including reactive oxygen and nitrogen species, proteases and proinflammatory/cytotoxic cytokines. Therapeutics under investigation include corticosteroids, N-acetyl cysteine, which has both mucolytic and antioxidant properties, inducible nitric oxide synthase inhibitors, liposomes containing superoxide dismutase, catalase, and/or tocopherols, protease inhibitors, and cytokine antagonists such as anti-tumor necrosis factor (TNF)-α antibody and pentoxifylline. Antifibrotic and fibrinolytic treatments may also prove beneficial in ameliorating airway obstruction and lung remodeling. More speculative approaches include inhibitors of transient receptor potential channels, which regulate pulmonary epithelial cell membrane permeability, non-coding RNAs and mesenchymal stem cells. As mustards represent high priority chemical threat agents, identification of effective therapeutics for mitigating toxicity is highly significant.
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role of reactive nitrogen species generated via inducible nitric oxide synthase in Vesicant induced lung injury inflammation and altered lung functioning
Toxicology and Applied Pharmacology, 2012Co-Authors: Vasanthi R Sunil, Jeffrey D Laskin, Jianliang Shen, Kinal Patelvayas, Andrew J Gow, Debra L LaskinAbstract:Pulmonary toxicity induced by Vesicants is associated with oxidative stress. In the present studies we analyzed the role of reactive nitrogen species (RNS) generated via inducible nitric oxide synthase (iNOS) in lung injury and inflammation induced by Vesicants using 2-chloroethyl ethyl sulfide (CEES) as a model. C57Bl/6 (WT) and iNOS−/− mice were sacrificed 3 d or 14 d following intratracheal administration of CEES (6 mg/kg) or control. CEES intoxication resulted in transient (3 d) increases in bronchoalveolar lavage (BAL) cell and protein content in WT, but not iNOS−/− mice. This correlated with expression of Ym1, a marker of oxidative stress in alveolar macrophages and epithelial cells. In contrast, in iNOS−/− mice, Ym1 was only observed 14 d post exposure in enlarged alveolar macrophages, suggesting that they are alternatively activated. This is supported by findings that lung tumor necrosis factor and lipocalin Lcn2 expression, mediators involved in tissue repair were also upregulated at this time in iNOS−/− mice. Conversely, CEES-induced increases in the proinflammatory genes, monocyte chemotactic protein-1 and cyclooxygenase-2, were abrogated in iNOS−/− mice. In WT mice, CEES treatment also resulted in increases in total lung resistance and decreases in compliance in response to methacholine, effects blunted by loss of iNOS. These data demonstrate that RNS, generated via iNOS play a role in the pathogenic responses to CEES, augmenting oxidative stress and inflammation and suppressing tissue repair. Elucidating inflammatory mechanisms mediating Vesicant-induced lung injury is key to the development of therapeutics to treat mustard poisoning.
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role of tnfr1 in lung injury and altered lung function induced by the model sulfur mustard Vesicant 2 chloroethyl ethyl sulfide
Toxicology and Applied Pharmacology, 2011Co-Authors: Vasanthi R Sunil, Jeffrey D Laskin, Jianliang Shen, Kinal Patelvayas, Andrew J Gow, Debra L LaskinAbstract:Lung toxicity induced by sulfur mustard is associated with inflammation and oxidative stress. To elucidate mechanisms mediating pulmonary damage, we used 2-chloroethyl ethyl sulfide (CEES), a model sulfur mustard Vesicant. Male mice (B6129) were treated intratracheally with CEES (3 or 6 mg/kg) or control. Animals were sacrificed 3, 7 or 14 days later and bronchoalveolar lavage (BAL) fluid and lung tissue collected. Treatment of mice with CEES resulted in an increase in BAL protein, an indication of alveolar epithelial damage, within 3 days. Expression of Ym1, an oxidative stress marker also increased in the lung, along with inducible nitric oxide synthase, and at 14 days, cyclooxygenase-2 and monocyte chemotactic protein-1, inflammatory proteins implicated in tissue injury. These responses were attenuated in mice lacking the p55 receptor for TNFα (TNFR1-/-), demonstrating that signaling via TNFR1 is key to CEES-induced injury, oxidative stress, and inflammation. CEES-induced upregulation of CuZn-superoxide dismutase (SOD) and MnSOD was delayed or absent in TNFR1-/- mice, relative to WT mice, suggesting that TNFα mediates early antioxidant responses to lung toxicants. Treatment of WT mice with CEES also resulted in functional alterations in the lung including decreases in compliance and increases in elastance. Additionally, methacholine-induced alterations in total lung resistance and central airway resistance were dampened by CEES. Loss of TNFR1 resulted in blunted functional responses to CEES. These effects were most notable in the airways. These data suggest that targeting TNFα signaling may be useful in mitigating lung injury, inflammation and functional alterations induced by Vesicants.
Donald R Gerecke - One of the best experts on this subject based on the ideXlab platform.
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expression of laminin 332 in Vesicant skin injury and wound repair
Clinical dermatology (Wilmington Del.), 2018Co-Authors: Yokechen Chang, Marion K Gordon, Donald R GereckeAbstract:Sulfur Mustard (SM) is a potent Vesicant or blistering agent. It is a highly reactive bi-functional alkylating agent that cross links proteins, DNA, and other cellular components. Laminin 332 is a heterotrimer glycoprotein and a crucial skin component that attaches the epidermal basal keratinocytes to the dermis. SM wounds histologically appear similar to Epidermolysis Bullosa (EB), human genetic blistering diseases that involve genetic changes in laminin 332. The specific mechanism of action of SM exposure is unknown, but there are several key similarities between Vesicant induced cutaneous injury and the Junctional form of EB (JEB) cutaneous injury: 1) Initial alkylation causes blistering similar to JEB; 2) Initial injury is followed by protease activation and prolonged inflammation similar to the chronic inflammation observed in EB; 3) The blister plane is at the level of the lamina lucida in the Basement Membrane Zone (BMZ) for both JEB and SM-induced injury. This suggests that injury induced by Vesicants is not unique and probably involves malformation of laminin 332. Understanding the role of laminin 332 in SM induced blisters may provide perspectives for future molecular therapeutic countermeasures against SM exposure.
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increased expression of the endocannabinoid system in mouse skin following exposure to sulfur mustard and nitrogen mustard mechlorethamine
The FASEB Journal, 2016Co-Authors: Diane E Heck, Donald R Gerecke, Debra L Laskin, Robert P Casillas, Irene Wohlman, Gabriella M Composto, Ned D Heindel, Laurie B Joseph, Jeffrey D LaskinAbstract:Vesicants including sulfur mustard (SM, bis(2-chloroethyl) sulfide) and nitrogen mustard (NM, bis(2-chloroethyl)methylamine) are highly reactive bifunctional alkylating agents that target the skin....
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sulfur mustard induces an endoplasmic reticulum stress response in the mouse ear Vesicant model
Toxicology and Applied Pharmacology, 2013Co-Authors: Yokechen Chang, Marion K Gordon, Jeffrey D Laskin, Robert P Casillas, James D Wang, Kathy K H Svoboda, Donald R GereckeAbstract:The endoplasmic reticulum (ER) stress response is a cell survival pathway upregulated when cells are under severe stress. Severely damaged mouse ear skin exposed to the Vesicant, sulfur mustard (bis-2-chloroethyl sulfide, SM), resulted in increased expression of ER chaperone proteins that accompany misfolded and incorrectly made proteins targeted for degradation. Time course studies with SM using the mouse ear Vesicant model (MEVM) showed progressive histopathologic changes including edema, separation of the epidermis from the dermis, persistent inflammation, upregulation of laminin γ2 (one of the chains of laminin-332, a heterotrimeric skin glycoprotein required for wound repair), and delayed wound healing from 24 h to 168 h post exposure. This was associated with time related increased expression of the cell survival ER stress marker, GRP78/BiP, and the ER stress apoptosis marker, GADD153/CHOP, suggesting simultaneous activation of both cell survival and non-mitochondrial apoptosis pathways. Dual immunofluorescence labeling of a keratinocyte migration promoting protein, laminin γ2 and GRP78/BIP, showed colocalization of the two molecules 72 h post exposure indicating that the laminin γ2 was misfolded after SM exposure and trapped within the ER. Taken together, these data show that ER stress is induced in mouse skin within 24 h of Vesicant exposure in a defensive response to promote cell survival; however, it appears that this response is rapidly overwhelmed by the apoptotic pathway as a consequence of severe SM-induced injury.
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regulation of hsp27 and hsp70 expression in human and mouse skin construct models by caveolae following exposure to the model sulfur mustard Vesicant 2 chloroethyl ethyl sulfide
Toxicology and Applied Pharmacology, 2011Co-Authors: Adrienne T Black, Donald R Gerecke, Diane E Heck, Debra L Laskin, Patrick J Hayden, Robert P Casillas, Patrick J Sinko, Jeffrey D LaskinAbstract:Dermal exposure to the Vesicant sulfur mustard causes marked inflammation and tissue damage. Basal keratinocytes appear to be a major target of sulfur mustard. In the present studies, mechanisms mediating skin toxicity were examined using a mouse skin construct model and a full-thickness human skin equivalent (EpiDerm-FT™). In both systems, administration of the model sulfur mustard Vesicant, 2-chloroethyl ethyl sulfide (CEES, 100-1000μM) at the air surface induced mRNA and protein expression of heat shock proteins 27 and 70 (Hsp27 and Hsp70). CEES treatment also resulted in increased expression of caveolin-1, the major structural component of caveolae. Immunohistochemistry revealed that Hsp27, Hsp70 and caveolin-1 were localized in basal and suprabasal layers of the epidermis. Caveolin-1 was also detected in fibroblasts in the dermal component of the full thickness human skin equivalent. Western blot analysis of caveolar membrane fractions isolated by sucrose density centrifugation demonstrated that Hsp27 and Hsp70 were localized in caveolae. Treatment of mouse keratinocytes with filipin III or methyl-β-cyclodextrin, which disrupt caveolar structure, markedly suppressed CEES-induced Hsp27 and Hsp70 mRNA and protein expression. CEES treatment is known to activate JNK and p38 MAP kinases; in mouse keratinocytes, inhibition of these enzymes suppressed CEES-induced expression of Hsp27 and Hsp70. These data suggest that MAP kinases regulate Hsp 27 and Hsp70; moreover, caveolae-mediated regulation of heat shock protein expression may be important in the pathophysiology of Vesicant-induced skin toxicity.
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expression of proliferative and inflammatory markers in a full thickness human skin equivalent following exposure to the model sulfur mustard Vesicant 2 chloroethyl ethyl sulfide
Toxicology and Applied Pharmacology, 2010Co-Authors: Adrienne T Black, Donald R Gerecke, Diane E Heck, Debra L Laskin, Patrick J Hayden, Robert P Casillas, Patrick J Sinko, Jeffrey D LaskinAbstract:a r t i c l e i n f o Article history: Sulfur mustard is a potent Vesicant that induces in! ammation, edema and blistering following dermal exposure. To assess molecular mechanisms mediating these responses, we analyzed the effects of the model sulfur mustard Vesicant, 2-chloroethyl ethyl sul" de, on EpiDerm-FT™, a commercially available full-thickness human skin equivalent. CEES (100-1000 ! M) caused a concentration-dependent increase in pyknotic nuclei and vacuolization in basal keratinocytes; at high concentrations (300-1000 ! M), CEES also disrupted keratin " lament architecture in the stratum corneum. This was associated with time-dependent increases in expression of proliferating cell nuclear antigen, a marker of cell proliferation, and poly(ADP-ribose) polymerase (PARP) and phosphorylated histone H2AX, markers of DNA damage. Concentration- and time- dependent increases in mRNA and protein expression of eicosanoid biosynthetic enzymes including COX-2, 5- lipoxygenase, microsomal PGE2 synthases, leukotriene (LT) A4 hydrolase and LTC4 synthase were observed in CEES-treated skin equivalents, as well as in antioxidant enzymes, glutathione S-transferases A1-2 (GSTA1-2), GSTA3 and GSTA4. These data demonstrate that CEES induces rapid cellular damage, cytotoxicity and in! ammation in full-thickness skin equivalents. These effects are similar to human responses to Vesicants in vivo and suggest that the full thickness skin equivalent is a useful in vitro model to characterize the biological effects of mustards and to develop potential therapeutics.