In Vitro Model of the Epidermis

Arnette, Christopher R.; Koetsier, Jennifer L.; Hoover, Paul; Getsios, Spiro; Green, Kathleen J.

doi:10.1016/bs.mie.2015.07.015

Cited by 59 publications

(38 citation statements)

References 40 publications

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“…Importantly, this 3D tissue culture system is removed from any systemic hormone influence and is maintained in a growth medium that typically is supplemented with exogenous hydrocortisone (1000nM) (Arnette et al , 2016). Using this model, we found an increase in HSD11B1 and HSD11B2 mRNA expression with progression of epidermal differentiation (p<0.05) (Supplementary Figure 6) that strongly correlated with markers of differentiation (loricrin, r=0.55, p<0.003).…”

Section: Resultsmentioning

confidence: 99%

Endogenous Glucocorticoid Deficiency in Psoriasis Promotes Inflammation and Abnormal Differentiation

Sarkar

Kaplan

Tsoi

et al. 2017

Journal of Investigative Dermatology

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The factors involved in maintaining a localized inflammatory state in psoriatic skin remain poorly understood. Here, we demonstrate through metabolomic and transcriptomic profiling marked suppression of glucocorticoid biosynthesis in the epidermis of psoriatic skin leading to localized deficiency of cortisol. Utilizing a 3D human epidermis model, we demonstrate that glucocorticoid biosynthesis is suppressed by pro-inflammatory cytokines and that glucocorticoid deficiency promotes inflammatory responses in keratinocytes. Finally, we show in vitro and in vivo that treatment with topical glucocorticoids leads to rapid restoration of glucocorticoid biosynthesis gene expression coincident with normalization of epidermal differentiation and suppression of inflammatory responses. Taken together, our data suggest that localized glucocorticoid deficiency in psoriatic skin interferes with epidermal differentiation and promotes a sustained and localized inflammatory response. This may shed new light on the mechanism of action of topical steroids, and demonstrates the critical role of endogenous steroid in maintaining both inflammatory and differentiation homeostasis in the epidermis.

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Section: Resultsmentioning

confidence: 99%

Endogenous Glucocorticoid Deficiency in Psoriasis Promotes Inflammation and Abnormal Differentiation

Sarkar

Kaplan

Tsoi

et al. 2017

Journal of Investigative Dermatology

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“…For 3-dimensional organotypic cultures, cells were grown to confluence in E-medium and grown at the air-medium interface for 12 days as previously described (Arnette et al, 2016, Meyers and Laimins, 1994). …”

Section: Methodsmentioning

confidence: 99%

PFKFB3, a Direct Target of p63, Is Required for Proliferation and Inhibits Differentiation in Epidermal Keratinocytes

Hamanaka

Mutlu

2017

Journal of Investigative Dermatology

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p63 is a transcription factor essential for epidermal development and homeostasis. p63 is a member of the p53 family of transcription factors, which are increasingly understood to be regulators of cellular metabolism. How p63 regulates metabolism in epidermal keratinocytes is incompletely understood, and it is unknown whether glycolytic regulation is essential to maintain the balance between proliferation and differentiation within the epidermis. We found that p63 promotes glycolytic metabolism in epidermal keratinocytes. p63 bound to consensus sites within the PFKFB3 gene and was required for PFKFB3 mRNA and protein expression. PFKFB3 overexpression inhibited differentiation of keratinocytes while knockdown inhibited proliferation and increased the rate of differentiation. Furthermore, we found that PFKFB3 was highly expressed in psoriatic epidermis. Our results reveal that PFKFB3 is a key regulator of epidermal homeostasis and may represent a therapeutic target for epidermal diseases associated with hyperproliferation and impaired differentiation.

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“…One option to overcome this problem is the use of organotypic skin equivalents, which are bioengineered substitutes that mimic the human skin (Alepee et al 2014 ; Arnette et al 2016 ). Today, they are commonly used for two different applications: on the one hand, as skin replacement or graft in clinical routine and, on the other hand, as an alternative to animal models for drug permeability and toxicity testing.…”

Section: Introductionmentioning

confidence: 99%

“…In the cosmetic industry, they even have replaced animal testing in regard to skin irritation, erosion and photosensitization. Depending on the complexity of the models, they can be divided into two subgroups: (1) Epidermal or dermal equivalents: three-dimensional models that consist only of keratinocytes (epidermal) or fibroblasts (dermal) growing on a suitable surface, and (2) full-thickness skin equivalents: more advanced models consisting of an artificial fibroblast-containing dermis together with an epidermis built from keratinocytes on top (Arnette et al 2016 ; Mathes et al 2014 ). Basically, full-thickness skin equivalents are built by preparation of a dermal layer (a collagen matrix populated with fibroblasts) followed by seeding of keratinocytes on top (Fig.…”

Section: Introductionmentioning

confidence: 99%

Long-term exposure of immortalized keratinocytes to arsenic induces EMT, impairs differentiation in organotypic skin models and mimics aspects of human skin derangements

et al. 2017

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Arsenic is one of the most important human carcinogens and environmental pollutants. However, the evaluation of the underlying carcinogenic mechanisms is challenging due to the lack of suitable in vivo and in vitro models, as distinct interspecies differences in arsenic metabolism exist. Thus, it is of high interest to develop new experimental models of arsenic-induced skin tumorigenesis in humans. Consequently, aim of this study was to establish an advanced 3D model for the investigation of arsenic-induced skin derangements, namely skin equivalents, built from immortalized human keratinocytes (NHEK/SVTERT3-5). In contrast to spontaneously immortalized HACAT cells, NHEK/SVTERT3-5 cells more closely resembled the differentiation pattern of primary keratinocytes. With regard to arsenic, our results showed that while our new cell model was widely unaffected by short-time treatment (72 h) with low, non-toxic doses of ATO (0.05–0.25 µM), chronic exposure (6 months) resulted in distinct changes of several cell characteristics. Thus, we observed an increase in the G2 fraction of the cell cycle accompanied by increased nucleus size and uneven tubulin distribution. Moreover, cells showed strong signs of de-differentiation and upregulation of several epithelial-to-mesenchymal transition markers. In line with these effects, chronic contact to arsenic resulted in impaired skin-forming capacities as well as localization of ki67-positive (proliferating) cells at the upper layers of the epidermis; a condition termed Bowen’s disease. Finally, chronically arsenic-exposed cells were characterized by an increased tumorigenicity in SCID mice. Taken together, our study presents a new model system for the investigation of mechanisms underlying the tumor-promoting effects of chronic arsenic exposure.Electronic supplementary materialThe online version of this article (doi:10.1007/s00204-017-2034-6) contains supplementary material, which is available to authorized users.

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In Vitro Model of the Epidermis

Cited by 59 publications

References 40 publications

Endogenous Glucocorticoid Deficiency in Psoriasis Promotes Inflammation and Abnormal Differentiation

Endogenous Glucocorticoid Deficiency in Psoriasis Promotes Inflammation and Abnormal Differentiation

PFKFB3, a Direct Target of p63, Is Required for Proliferation and Inhibits Differentiation in Epidermal Keratinocytes

Long-term exposure of immortalized keratinocytes to arsenic induces EMT, impairs differentiation in organotypic skin models and mimics aspects of human skin derangements

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