caa a22 4500 445321962 CHVBK 20180317142707.0 cr unu---uuuuu 170323e20111001xx s 000 0 eng 10.1007/s10534-011-9441-6 doi (NATIONALLICENCE)springer-10.1007/s10534-011-9441-6 Zinc pyrithione impairs zinc homeostasis and upregulates stress response gene expression in reconstructed human epidermis [Elektronische Daten] [Sarah Lamore, Georg Wondrak] Zinc ion homeostasis plays an important role in human cutaneous biology where it is involved in epidermal differentiation and barrier function, inflammatory and antimicrobial regulation, and wound healing. Zinc-based compounds designed for topical delivery therefore represent an important class of cutaneous therapeutics. Zinc pyrithione (ZnPT) is an FDA-approved microbicidal agent used worldwide in over-the-counter topical antimicrobials, and has also been examined as an investigational therapeutic targeting psoriasis and UVB-induced epidermal hyperplasia. Recently, we have demonstrated that cultured primary human skin keratinocytes display an exquisite sensitivity to nanomolar ZnPT concentrations causing induction of heat shock response gene expression and poly(ADP-ribose) polymerase (PARP)-dependent cell death (Cell Stress Chaperones 15:309-322, 2010). Here we demonstrate that ZnPT causes rapid accumulation of intracellular zinc in primary keratinocytes as observed by quantitative fluorescence microscopy and inductively coupled plasma mass spectrometry (ICP-MS), and that PARP activation, energy crisis, and genomic impairment are all antagonized by zinc chelation. In epidermal reconstructs (EpiDerm™) exposed to topical ZnPT (0.1-2% in Vanicream™), ICP-MS demonstrated rapid zinc accumulation, and expression array analysis demonstrated upregulation of stress response genes encoding metallothionein-2A (MT2A), heat shock proteins (HSPA6, HSPA1A, HSPB5, HSPA1L, DNAJA1, HSPH1, HSPD1, HSPE1), antioxidants (SOD2, GSTM3, HMOX1), and the cell cycle inhibitor p21 (CDKN1A). IHC analysis of ZnPT-treated EpiDerm™ confirmed upregulation of Hsp70 and TUNEL-positivity. Taken together our data demonstrate that ZnPT impairs zinc ion homeostasis and upregulates stress response gene expression in primary keratinocytes and reconstructed human epidermis, activities that may underlie therapeutic and toxicological effects of this topical drug. Springer Science+Business Media, LLC., 2011 Zinc pyrithione nationallicence Keratinocyte nationallicence Reconstructed epidermis nationallicence Heat shock response nationallicence HSPA1A nationallicence ICP-MS nationallicence AV : AnnexinV nationallicence FITC : Fluorescein isothiocyanate nationallicence DTPA : Diethylenetriaminepentaacetic acid nationallicence HSP : Heat shock protein nationallicence ICP-MS : Inductively coupled plasma mass spectrometry nationallicence IHC : Immunohistochemistry nationallicence NHEK : Normal human epidermal keratinocyte nationallicence OTC : Over-the-counter nationallicence PAR : Poly(ADP-ribose) polymer nationallicence PARP : Poly(ADP-ribose) polymerase nationallicence PBS : Phosphate buffered saline nationallicence PI : Propidium iodide nationallicence SDS-PAGE : Sodium dodecylsulfate polyacrylamide gel electrophoresis nationallicence TPEN : N,N,N′,N′-tetrakis(2-pyridylmethyl)ethylenediamine nationallicence TUNEL : Terminal dUTP nick end labeling nationallicence ZnPT : Zinc pyrithione nationallicence Lamore Sarah Department of Pharmacology and Toxicology, College of Pharmacy and Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, 85724, Tucson, AZ, USA aut Wondrak Georg Department of Pharmacology and Toxicology, College of Pharmacy and Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, 85724, Tucson, AZ, USA aut BioMetals Springer Netherlands 24/5(2011-10-01), 875-890 0966-0844 24:5<875 2011 24 10534 https://doi.org/10.1007/s10534-011-9441-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10534-011-9441-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Lamore Sarah Department of Pharmacology and Toxicology, College of Pharmacy and Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, 85724, Tucson, AZ, USA aut NATIONALLICENCE 700 1- Wondrak Georg Department of Pharmacology and Toxicology, College of Pharmacy and Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, 85724, Tucson, AZ, USA aut NATIONALLICENCE 773 0- BioMetals Springer Netherlands 24/5(2011-10-01), 875-890 0966-0844 24:5<875 2011 24 10534 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer