PMID- 33635393 OWN - NLM STAT- MEDLINE DCOM- 20211103 LR - 20211103 IS - 1432-0738 (Electronic) IS - 0340-5761 (Print) IS - 0340-5761 (Linking) VI - 95 IP - 4 DP - 2021 Apr TI - Alkylated epidermal creatine kinase as a biomarker for sulfur mustard exposure: comparison to adducts of albumin and DNA in an in vivo rat study. PG - 1323-1333 LID - 10.1007/s00204-021-03005-3 [doi] AB - Sulfur mustard (SM) is a chemical warfare agent which use is banned under international law and that has been used recently in Northern Iraq and Syria by the so-called Islamic State. SM induces the alkylation of endogenous proteins like albumin and hemoglobin thus forming covalent adducts that are targeted by bioanalytical methods for the verification of systemic poisoning. We herein report a novel biomarker, namely creatine kinase (CK) B-type, suitable as a local biomarker for SM exposure on the skin. Human and rat skin were proven to contain CK B-type by Western blot analysis. Following exposure to SM ex vivo, the CK-adduct was extracted from homogenates by immunomagnetic separation and proteolyzed afterwards. The cysteine residue Cys(282) was found to be alkylated by the SM-specific hydroxyethylthioethyl (HETE)-moiety detected as the biomarker tetrapeptide TC(-HETE)PS. A selective and sensitive micro liquid chromatography-electrospray ionization high-resolution tandem-mass spectrometry (microLC-ESI MS/HRMS) method was developed to monitor local CK-adducts in an in vivo study with rats percutaneously exposed to SM. CK-adduct formation was compared to already established DNA- and systemic albumin biomarkers. CK- and DNA-adducts were successfully detected in biopsies of exposed rat skin as well as albumin-adducts in plasma. Relative biomarker concentrations make the CK-adduct highly appropriate as a local dermal biomarker. In summary, CK or rather Cys(282) in CK B-type was identified as a new, additional dermal target of local SM exposures. To our knowledge, it is also the first time that HETE-albumin adducts, and HETE-DNA adducts were monitored simultaneously in an in vivo animal study. FAU - Steinritz, Dirk AU - Steinritz D AUID- ORCID: 0000-0002-2073-5683 AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. dirk.steinritz@lrz.uni-muenchen.de. AD - Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universitat Munich (LMU), Goethestrasse 33 / Nussbaumstrasse 26, 80366, Munich, Germany. dirk.steinritz@lrz.uni-muenchen.de. AD - Bundeswehr Medical Service Academy, Ingolstadter Strasse 240, 80939, Munich, Germany. dirk.steinritz@lrz.uni-muenchen.de. FAU - Luling, Robin AU - Luling R AUID- ORCID: 0000-0002-6392-2542 AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. AD - Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universitat Munich (LMU), Goethestrasse 33 / Nussbaumstrasse 26, 80366, Munich, Germany. FAU - Siegert, Markus AU - Siegert M AUID- ORCID: 0000-0003-1760-7733 AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. AD - Proteros Biostrucures GmbH, Bunsenstrasse 7a, 82152, Planegg, Germany. FAU - Herbert, Julia AU - Herbert J AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. AD - Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Rutgers University, Piscataway, NJ, USA. FAU - Muckter, Harald AU - Muckter H AD - Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universitat Munich (LMU), Goethestrasse 33 / Nussbaumstrasse 26, 80366, Munich, Germany. FAU - Taeger, Christian D AU - Taeger CD AD - Department of Plastic-, Hand- and Reconstructive Surgery, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053, Regensburg, Germany. FAU - Gudermann, Thomas AU - Gudermann T AD - Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universitat Munich (LMU), Goethestrasse 33 / Nussbaumstrasse 26, 80366, Munich, Germany. FAU - Dietrich, Alexander AU - Dietrich A AUID- ORCID: 0000-0002-1168-8707 AD - Walther-Straub-Institute of Pharmacology and Toxicology, Ludwig-Maximilians-Universitat Munich (LMU), Goethestrasse 33 / Nussbaumstrasse 26, 80366, Munich, Germany. FAU - Thiermann, Horst AU - Thiermann H AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. FAU - John, Harald AU - John H AUID- ORCID: 0000-0002-7126-6709 AD - Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937, Munich, Germany. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20210226 PL - Germany TA - Arch Toxicol JT - Archives of toxicology JID - 0417615 RN - 0 (Albumins) RN - 0 (Biomarkers) RN - 0 (Chemical Warfare Agents) RN - 0 (DNA Adducts) RN - EC 2.7.3.2 (Creatine Kinase) RN - K848JZ4886 (Cysteine) RN - T8KEC9FH9P (Mustard Gas) SB - IM MH - Albumins/metabolism MH - Alkylation/drug effects MH - Animals MH - Biomarkers/metabolism MH - Chemical Warfare Agents/*toxicity MH - Chromatography, Liquid MH - Creatine Kinase/*metabolism MH - Cysteine/metabolism MH - DNA Adducts/metabolism MH - Humans MH - Male MH - Mustard Gas/*toxicity MH - Rats MH - Rats, Wistar MH - Skin/*drug effects/metabolism MH - Spectrometry, Mass, Electrospray Ionization MH - Tandem Mass Spectrometry PMC - PMC8032612 OTO - NOTNLM OT - Chemical warfare agents OT - High-resolution tandem-mass spectrometry OT - Human skin OT - Micro liquid chromatography COIS- The authors declare no conflict of interest. EDAT- 2021/02/27 06:00 MHDA- 2021/11/04 06:00 PMCR- 2021/02/26 CRDT- 2021/02/26 12:12 PHST- 2020/11/27 00:00 [received] PHST- 2021/02/16 00:00 [accepted] PHST- 2021/02/27 06:00 [pubmed] PHST- 2021/11/04 06:00 [medline] PHST- 2021/02/26 12:12 [entrez] PHST- 2021/02/26 00:00 [pmc-release] AID - 10.1007/s00204-021-03005-3 [pii] AID - 3005 [pii] AID - 10.1007/s00204-021-03005-3 [doi] PST - ppublish SO - Arch Toxicol. 2021 Apr;95(4):1323-1333. doi: 10.1007/s00204-021-03005-3. Epub 2021 Feb 26.