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The topical antimicrobial zinc pyrithione is a heat shock response inducer that causes DNA damage and PARP-dependent energy crisis in human skin cells

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Cell Stress and Chaperones Aims and scope

Abstract

The differentiated epidermis of human skin serves as an essential barrier against environmental insults from physical, chemical, and biological sources. Zinc pyrithione (ZnPT) is an FDA-approved microbicidal agent used worldwide in clinical antiseptic products, over-the-counter topical antimicrobials, and cosmetic consumer products including antidandruff shampoos. Here we demonstrate for the first time that cultured primary human skin keratinocytes and melanocytes display an exquisite vulnerability to nanomolar concentrations of ZnPT resulting in pronounced induction of heat shock response gene expression and impaired genomic integrity. In keratinocytes treated with nanomolar concentrations of ZnPT, expression array analysis revealed massive upregulation of genes encoding heat shock proteins (HSPA6, HSPA1A, HSPB5, HMOX1, HSPA1L, and DNAJA1) further confirmed by immunodetection. Moreover, ZnPT treatment induced rapid depletion of cellular ATP levels and formation of poly(ADP-ribose) polymers. Consistent with an involvement of poly(ADP-ribose) polymerase (PARP) in ZnPT-induced energy crisis, ATP depletion could be antagonized by pharmacological inhibition of PARP. This result was independently confirmed using PARP-1 knockout mouse embryonic fibroblasts that were resistant to ATP depletion and cytotoxicity resulting from ZnPT exposure. In keratinocytes and melanocytes, single-cell gel electrophoresis and flow cytometric detection of γ-H2A.X revealed rapid induction of DNA damage in response to ZnPT detectable before general loss of cell viability occurred through caspase-independent pathways. Combined with earlier experimental evidence that documents penetration of ZnPT through mammalian skin, our findings raise the possibility that this topical antimicrobial may target and compromise keratinocytes and melanocytes in intact human skin.

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Abbreviations

3-ABA:

3-Aminobenzamide

AV:

AnnexinV

FITC:

Fluorescein isothiocyanate

DTPA:

Diethylenetriaminepentaacetic acid

EGR1:

Early growth response protein 1

Fpg:

Formamidopyrimidine-glycosylase

HEK:

Human epidermal keratinocyte

HEM:

Human epidermal melanocyte

HO-1:

Heme oxygenase 1

HSP:

Heat shock protein

MEF:

Mouse embryonal fibroblast

NAC:

N-Acetyl-l-cysteine

OTC:

Over-the-counter

PAR:

Poly(ADP-ribose) polymer

PARP:

Poly(ADP-ribose) polymerase

PI:

Propidium iodide

SDS-PAGE:

Sodium dodecylsulfate polyacrylamide gel electrophoresis

ZnPT:

Zinc pyrithione

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Acknowledgments

Supported in part by grants from the National Institutes of Health (Award Number R01CA122484 from the National Cancer Institute, ES007091, ES06694, Arizona Cancer Center Support Grant CA023074), NSF (DGE-0114420 to CMC), and the Arizona Science Foundation (to SL). The content is solely the responsibility of the author and does not necessarily represent the official views of the National Cancer Institute or the National Institutes of Health.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, College of Pharmacy, Arizona Cancer Center, University of Arizona, Tucson, AZ, USA

    Sarah D. Lamore, Christopher M. Cabello & Georg T. Wondrak

  2. College of Pharmacy & Arizona Cancer Center, University of Arizona, 1515 North Campbell Avenue, Tucson, AZ, 85724, USA

    Georg T. Wondrak

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  1. Sarah D. Lamore
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  2. Christopher M. Cabello
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  3. Georg T. Wondrak
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Correspondence to Georg T. Wondrak.

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Lamore, S.D., Cabello, C.M. & Wondrak, G.T. The topical antimicrobial zinc pyrithione is a heat shock response inducer that causes DNA damage and PARP-dependent energy crisis in human skin cells. Cell Stress and Chaperones 15, 309–322 (2010). https://doi.org/10.1007/s12192-009-0145-6

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  • DOI: https://doi.org/10.1007/s12192-009-0145-6

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