A new in vitro method for identifying chemical sensitizers combining peptide binding with ARE/EpRE-mediated gene expression in human skin cells.
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McKim JM Jr, Keller DJ 3rd, Gorski JR
A new in vitro method for identifying chemical sensitizers combining peptide binding with ARE/EpRE-mediated gene expression in human skin cells.
Cutan Ocul Toxicol. 2010 Sep;29(3):171-92. doi: 10.3109/15569527.2010.483869.
- PubMed ID
- 20491607 [ View in PubMed]
- Abstract
Allergic contact dermatitis (ACD) is a significant safety concern for developers of cosmetic, personal care, chemical, pharmaceutical, and medical device products. The guinea pig maximization test (GMPT) and the murine local lymph node assay (LLNA) are accepted methods for determining chemical sensitization. Recent legislative initiatives in Europe require the development of new in vitro alternatives to animal tests for chemical sensitization. The aim of this project was to develop an in vitro screening method that uses a human skin cell line (HaCaT), chemical reactivity, and gene expression profiling to identify positive and negative responses, to place chemicals into potency categories of extreme/strong (ES), moderate (M), weak (W), and nonsensitizers (N), and to provide an estimate of corresponding LLNA values. The method and processing algorithm were developed from a training set of 39 chemicals possessing a wide range of sensitization potencies. Three cationic metals, chromium (Cr), nickel (Ni), and silver (Ag), were also evaluated in this model. Chemical reactivity was determined by measuring glutathione (GSH) depletion in a cell free matrix. Three signaling pathways (Keap1/Nrf 2/ARE/EpRE, ARNT/AhR/XRE, and Nrf1/MTF/MRE) that are known to be activated by sensitizing agents were monitored by measuring the relative abundance of 11 genes whose expression is controlled by one of these 3 pathways. Final exposure concentrations were based on toxicity and solubility. A range-finding experiment was conducted with each compound to determine cytotoxicity and solubility. Six exposure concentrations (0.1 to 2,500 microM) and an exposure time of 24 hours were used in the final experiments. Glutathione depletion alone did not provide the accuracy necessary to differentiate potency categories. However, chemical reactivity combined with gene expression profiles significantly improved the in vitro predictions. A predicted toxicity index (PTI) was determined for each test chemical. A comparison of LLNA values with PTI values revealed an inverse relationship. The large variation in LLNA data for compounds in the same potency category makes direct extrapolation from PTI to LLNA difficult. To challenge the system, 58 additional compounds were submitted in a blinded manner. Compounds placed into ES and M categories were considered positive, whereas compounds classified as W or N were considered negative. Accuracy was approximately 84%, with a sensitivity of 81% and a specificity of 92%. The model correctly identified 2 of 3 cationic metals as positive. In conclusion, the method described here demonstrates a valuable in vitro method for identifying chemicals and metals that induce skin sensitization.
DrugBank Data that Cites this Article
- Pharmaco-transcriptomics
Drug Drug Groups Gene Gene ID Change Interaction Chromosome 2-Aminophenol Experimental AKR1C2 1646 upregulated 2-aminophenol results in increased expression of AKR1C2 mRNA 10p15.1 2-Aminophenol Experimental CXCL8 3576 upregulated 2-aminophenol results in increased expression of CXCL8 mRNA 4q13.3 2-Aminophenol Experimental CYP1A1 1543 upregulated 2-aminophenol results in increased expression of CYP1A1 mRNA 15q24.1 2-Aminophenol Experimental HMOX1 3162 upregulated 2-aminophenol results in increased expression of HMOX1 mRNA 22q12.3 2-Aminophenol Experimental NQO1 1728 upregulated 2-aminophenol results in increased expression of NQO1 mRNA 16q22.1 Benzoic acid Approved Investigational MT1A 4489 upregulated Benzoic Acid results in increased expression of MT1A mRNA 16q13 Benzoic acid Approved Investigational MT2A 4502 upregulated Benzoic Acid results in increased expression of MT2A mRNA 16q13 Benzoic acid Approved Investigational TXN 7295 upregulated Benzoic Acid results in increased expression of TXN mRNA 9q31.3 2-mercaptobenzothiazole Approved Experimental Vet Approved AKR1C2 1646 upregulated captax results in increased expression of AKR1C2 mRNA 10p15.1 2-mercaptobenzothiazole Approved Experimental Vet Approved CYP1A1 1543 upregulated captax results in increased expression of CYP1A1 mRNA 15q24.1 2-mercaptobenzothiazole Approved Experimental Vet Approved GCLC 2729 upregulated captax results in increased expression of GCLC mRNA 6p12.1 Dinitrochlorobenzene Investigational AKR1C2 1646 upregulated Dinitrochlorobenzene results in increased expression of AKR1C2 mRNA 10p15.1 Dinitrochlorobenzene Investigational CXCL8 3576 upregulated Dinitrochlorobenzene results in increased expression of CXCL8 mRNA 4q13.3 Dinitrochlorobenzene Investigational CYP1A1 1543 upregulated Dinitrochlorobenzene results in increased expression of CYP1A1 mRNA 15q24.1 Dinitrochlorobenzene Investigational MAFF 23764 upregulated Dinitrochlorobenzene results in increased expression of MAFF mRNA 22q13.1 Dinitrochlorobenzene Investigational MT2A 4502 upregulated Dinitrochlorobenzene results in increased expression of MT2A mRNA 16q13 Dinitrochlorobenzene Investigational NQO1 1728 upregulated Dinitrochlorobenzene results in increased expression of NQO1 mRNA 16q22.1 Diphencyprone Investigational HMOX1 3162 upregulated diphenylcyclopropenone results in increased expression of HMOX1 mRNA 22q12.3 Diphencyprone Investigational MAFF 23764 upregulated diphenylcyclopropenone results in increased expression of MAFF mRNA 22q13.1 Diphencyprone Investigational MT1A 4489 upregulated diphenylcyclopropenone results in increased expression of MT1A mRNA 16q13 Diphencyprone Investigational MT2A 4502 upregulated diphenylcyclopropenone results in increased expression of MT2A mRNA 16q13 Diphencyprone Investigational TXN 7295 upregulated diphenylcyclopropenone results in increased expression of TXN mRNA 9q31.3 Formaldehyde Approved Vet Approved AKR1C2 1646 upregulated Formaldehyde results in increased expression of AKR1C2 mRNA 10p15.1 Formaldehyde Approved Vet Approved CYP1A1 1543 upregulated Formaldehyde results in increased expression of CYP1A1 mRNA 15q24.1 Formaldehyde Approved Vet Approved GCLC 2729 upregulated Formaldehyde results in increased expression of GCLC mRNA 6p12.1 Formaldehyde Approved Vet Approved TXN 7295 upregulated Formaldehyde results in increased expression of TXN mRNA 9q31.3 Glycerin Approved Investigational HMOX1 3162 upregulated Glycerol results in increased expression of HMOX1 mRNA 22q12.3 Glycerin Approved Investigational MT1A 4489 upregulated Glycerol results in increased expression of MT1A mRNA 16q13 Glycerin Approved Investigational MT2A 4502 upregulated Glycerol results in increased expression of MT2A mRNA 16q13 Phenylacetaldehyde Experimental AKR1C2 1646 upregulated phenylacetaldehyde results in increased expression of AKR1C2 mRNA 10p15.1 Propyl Gallate Investigational AKR1C2 1646 upregulated Propyl Gallate results in increased expression of AKR1C2 mRNA 10p15.1 Propyl Gallate Investigational CYP1A1 1543 upregulated Propyl Gallate results in increased expression of CYP1A1 mRNA 15q24.1 Propyl Gallate Investigational HMOX1 3162 upregulated Propyl Gallate results in increased expression of HMOX1 mRNA 22q12.3 Propyl Gallate Investigational MAFF 23764 upregulated Propyl Gallate results in increased expression of MAFF mRNA 22q13.1 Salicylic acid Approved Investigational Vet Approved AKR1C2 1646 upregulated Salicylic Acid results in increased expression of AKR1C2 mRNA 10p15.1 Salicylic acid Approved Investigational Vet Approved MAFF 23764 upregulated Salicylic Acid results in increased expression of MAFF mRNA 22q13.1 Salicylic acid Approved Investigational Vet Approved MT1A 4489 upregulated Salicylic Acid results in increased expression of MT1A mRNA 16q13 Salicylic acid Approved Investigational Vet Approved TXN 7295 upregulated Salicylic Acid results in increased expression of TXN mRNA 9q31.3 Silver nitrate Approved Investigational CXCL8 3576 upregulated Silver Nitrate results in increased expression of CXCL8 mRNA 4q13.3 Silver nitrate Approved Investigational MT2A 4502 upregulated Silver Nitrate results in increased expression of MT2A mRNA 16q13 Sodium lauryl sulfate Experimental AKR1C2 1646 upregulated Sodium Dodecyl Sulfate results in increased expression of AKR1C2 mRNA 10p15.1 Sodium lauryl sulfate Experimental CXCL8 3576 upregulated Sodium Dodecyl Sulfate results in increased expression of CXCL8 mRNA 4q13.3 Sodium lauryl sulfate Experimental CYP1A1 1543 upregulated Sodium Dodecyl Sulfate results in increased expression of CYP1A1 mRNA 15q24.1 Sodium lauryl sulfate Experimental MAFF 23764 upregulated Sodium Dodecyl Sulfate results in increased expression of MAFF mRNA 22q13.1 Sodium lauryl sulfate Experimental MT1A 4489 upregulated Sodium Dodecyl Sulfate results in increased expression of MT1A mRNA 16q13 Sodium lauryl sulfate Experimental NQO1 1728 upregulated Sodium Dodecyl Sulfate results in increased expression of NQO1 mRNA 16q22.1 Sodium lauryl sulfate Experimental TXN 7295 upregulated Sodium Dodecyl Sulfate results in increased expression of TXN mRNA 9q31.3