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Article ; Online: Skin microbiome and antimicrobial peptides.

Zeeuwen, Patrick L J M / Grice, Elizabeth A

2021 Volume 30, Issue 10, Page(s) 1362–1365

MeSH term(s)	Antimicrobial Peptides/immunology ; Humans ; Microbiota/immunology ; Skin/immunology ; Skin/microbiology
Chemical Substances	Antimicrobial Peptides
Language	English
Publishing date	2021-09-01
Publishing country	Denmark
Document type	Editorial ; Introductory Journal Article
ZDB-ID	1130936-2
ISSN	1600-0625 ; 0906-6705
ISSN (online)	1600-0625
ISSN	0906-6705
DOI	10.1111/exd.14454
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Article ; Online: Keratinocyte signaling in atopic dermatitis: Investigations in organotypic skin models toward clinical application.

Meesters, Luca D / Niehues, Hanna / Johnston, Luke / Smits, Jos P H / Zeeuwen, Patrick L J M / Brown, Sara J / van den Bogaard, Ellen H

The Journal of allergy and clinical immunology

2023 Volume 151, Issue 5, Page(s) 1231–1235

MeSH term(s)	Humans ; Dermatitis, Atopic ; Skin ; Keratinocytes ; Signal Transduction
Language	English
Publishing date	2023-02-23
Publishing country	United States
Document type	Editorial ; Research Support, Non-U.S. Gov't
ZDB-ID	121011-7
ISSN	1097-6825 ; 1085-8725 ; 0091-6749
ISSN (online)	1097-6825 ; 1085-8725
ISSN	0091-6749
DOI	10.1016/j.jaci.2023.02.012
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Article ; Online: The aryl hydrocarbon receptor regulates epidermal differentiation through transient activation of TFAP2A.

Smits, Jos P H / Qu, Jieqiong / Pardow, Felicitas / van den Brink, Noa J M / Rodijk-Olthuis, Diana / van Vlijmen-Willems, Ivonne M J J / van Heeringen, Simon J / Zeeuwen, Patrick L J M / Schalkwijk, Joost / Zhou, Huiqing / van den Bogaard, Ellen H

The Journal of investigative dermatology

2024

Abstract: The aryl hydrocarbon receptor (AHR) is an evolutionary conserved environmental sensor identified as indispensable regulator of epithelial homeostasis and barrier organ function. Molecular signaling cascade and target genes upon AHR activation and their ... ...

Abstract	The aryl hydrocarbon receptor (AHR) is an evolutionary conserved environmental sensor identified as indispensable regulator of epithelial homeostasis and barrier organ function. Molecular signaling cascade and target genes upon AHR activation and their contribution to cell and tissue function are however not fully understood. Multi-omics analyses using human skin keratinocytes revealed that, upon ligand activation, AHR binds open chromatin to induce expression of transcription factors (TFs), e.g., Transcription Factor AP-2α (TFAP2A), as a swift response to environmental stimuli. The terminal differentiation program including upregulation of barrier genes, filaggrin and keratins, was mediated by TFAP2A as a secondary response to AHR activation. The role of AHR-TFAP2A axis in controlling keratinocyte terminal differentiation for proper barrier formation was further confirmed using CRISPR/Cas9 in human epidermal equivalents. Overall, the study provides additional insights into the molecular mechanism behind AHR-mediated barrier function and identifies potential targets for the treatment of skin barrier diseases.
Language	English
Publishing date	2024-02-22
Publishing country	United States
Document type	Journal Article
ZDB-ID	80136-7
ISSN	1523-1747 ; 0022-202X
ISSN (online)	1523-1747
ISSN	0022-202X
DOI	10.1016/j.jid.2024.01.030
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Article ; Online: The skin barrier: Epidermis vs environment.

Eckhart, Leopold / Zeeuwen, Patrick L J M

Experimental dermatology

2018 Volume 27, Issue 8, Page(s) 805–806

MeSH term(s)	Animals ; Dermatology/methods ; Environment ; Epidermis/immunology ; Epidermis/physiology ; Humans ; Inflammation ; Keratinocytes/cytology ; Skin/immunology ; Skin Diseases/prevention & control ; Skin Diseases/therapy ; Skin Physiological Phenomena
Language	English
Publishing date	2018-08-09
Publishing country	Denmark
Document type	Editorial ; Introductory Journal Article
ZDB-ID	1130936-2
ISSN	1600-0625 ; 0906-6705
ISSN (online)	1600-0625
ISSN	0906-6705
DOI	10.1111/exd.13731
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Article ; Online: Gram-positive anaerobic cocci guard skin homeostasis by regulating host-defense mechanisms.

van der Krieken, Danique A / Rikken, Gijs / Ederveen, Thomas H A / Jansen, Patrick A M / Rodijk-Olthuis, Diana / Meesters, Luca D / van Vlijmen-Willems, Ivonne M J J / van Cranenbroek, Bram / van der Molen, Renate G / Schalkwijk, Joost / van den Bogaard, Ellen H / Zeeuwen, Patrick L J M

iScience

2023 Volume 26, Issue 4, Page(s) 106483

Abstract: In atopic dermatitis (AD), chronic skin inflammation is associated with skin barrier defects and skin microbiome dysbiosis including a lower abundance of Gram-positive anaerobic cocci (GPACs). We here report that, through secreted soluble factors, GPAC ... ...

Abstract	In atopic dermatitis (AD), chronic skin inflammation is associated with skin barrier defects and skin microbiome dysbiosis including a lower abundance of Gram-positive anaerobic cocci (GPACs). We here report that, through secreted soluble factors, GPAC rapidly and directly induced epidermal host-defense molecules in cultured human keratinocytes and indirectly via immune-cell activation and cytokines derived thereof. Host-derived antimicrobial peptides known to limit the growth of
Language	English
Publishing date	2023-03-23
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.106483
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Article ; Online: CYSRT1: An Antimicrobial Epidermal Protein that Can Interact with Late Cornified Envelope Proteins.

Niehues, Hanna / Rikken, Gijs / Kersten, Ferry F J / Eeftens, Jorine M / van Vlijmen-Willems, Ivonne M J J / Rodijk-Olthuis, Diana / Jansen, Patrick A M / Hendriks, Wiljan J A J / Ederveen, Thomas H A / Schalkwijk, Joost / van den Bogaard, Ellen H / Zeeuwen, Patrick L J M

The Journal of investigative dermatology

2023 Volume 143, Issue 8, Page(s) 1498–1508.e7

Abstract: Late cornified envelope (LCE) proteins are small cationic epidermal proteins with antimicrobial properties, and the combined deletion of LCE3B and LCE3C genes is a risk factor for psoriasis that affects skin microbiome composition. In a yeast two-hybrid ... ...

Abstract	Late cornified envelope (LCE) proteins are small cationic epidermal proteins with antimicrobial properties, and the combined deletion of LCE3B and LCE3C genes is a risk factor for psoriasis that affects skin microbiome composition. In a yeast two-hybrid screen, we identified CYSRT1 as an interacting partner of members of all LCE groups except LCE6. These interactions were confirmed in a mammalian cell system by coimmunoprecipitation. CYSRT1 is a protein of unknown function that is specifically expressed in cutaneous and oral epithelia and spatially colocalizes with LCE proteins in the upper layers of the suprabasal epidermis. Constitutive CYSRT1 expression is present in fully differentiated epidermis and can be further induced in vivo by disruption of the skin barrier upon stratum corneum removal. Transcriptional regulation correlates to keratinocyte terminal differentiation but not to skin bacteria exposure. Similar to LCEs, CYSRT1 was found to have antibacterial activity against Pseudomonas aeruginosa. Comparative gene sequence analysis and protein amino acid alignment indicate that CYSRT1 is highly conserved among vertebrates and has putative antimicrobial activity. To summarize, we identified CYSRT1 in the outer skin layer, where it colocalizes with LCE proteins and contributes to the constitutive epidermal antimicrobial host defense repertoire.
MeSH term(s)	Anti-Infective Agents/metabolism ; Cornified Envelope Proline-Rich Proteins/genetics ; Cornified Envelope Proline-Rich Proteins/metabolism ; Epidermis/metabolism ; Keratinocytes/metabolism ; Proteins/metabolism ; Psoriasis/genetics ; Psoriasis/metabolism ; Skin/metabolism ; Humans
Chemical Substances	Anti-Infective Agents ; Cornified Envelope Proline-Rich Proteins ; Proteins
Language	English
Publishing date	2023-02-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80136-7
ISSN	1523-1747 ; 0022-202X
ISSN (online)	1523-1747
ISSN	0022-202X
DOI	10.1016/j.jid.2023.01.022
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Article ; Online: Novel methodologies for host-microbe interactions and microbiome-targeted therapeutics in 3D organotypic skin models.

Rikken, Gijs / Meesters, Luca D / Jansen, Patrick A M / Rodijk-Olthuis, Diana / van Vlijmen-Willems, Ivonne M J J / Niehues, Hanna / Smits, Jos P H / Oláh, Peter / Homey, Bernhard / Schalkwijk, Joost / Zeeuwen, Patrick L J M / van den Bogaard, Ellen H

Microbiome

2023 Volume 11, Issue 1, Page(s) 227

Abstract: Background: Following descriptive studies on skin microbiota in health and disease, mechanistic studies on the interplay between skin and microbes are on the rise, for which experimental models are in great demand. Here, we present a novel methodology ... ...

Abstract	Background: Following descriptive studies on skin microbiota in health and disease, mechanistic studies on the interplay between skin and microbes are on the rise, for which experimental models are in great demand. Here, we present a novel methodology for microbial colonization of organotypic skin and analysis thereof. Results: An inoculation device ensured a standardized application area on the stratum corneum and a homogenous distribution of bacteria, while preventing infection of the basolateral culture medium even during prolonged culture periods for up to 2 weeks at a specific culture temperature and humidity. Hereby, host-microbe interactions and antibiotic interventions could be studied, revealing diverse host responses to various skin-related bacteria and pathogens. Conclusions: Our methodology is easily transferable to a wide variety of organotypic skin or mucosal models and different microbes at every cell culture facility at low costs. We envision that this study will kick-start skin microbiome studies using human organotypic skin cultures, providing a powerful alternative to experimental animal models in pre-clinical research. Video Abstract.
MeSH term(s)	Animals ; Humans ; Host Microbial Interactions ; Skin/microbiology ; Epidermis ; Microbiota ; Models, Animal
Language	English
Publishing date	2023-10-17
Publishing country	England
Document type	Video-Audio Media ; Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2697425-3
ISSN	2049-2618 ; 2049-2618
ISSN (online)	2049-2618
ISSN	2049-2618
DOI	10.1186/s40168-023-01668-x
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Article: The aryl hydrocarbon receptor regulates epidermal differentiation through transient activation of TFAP2A.

bioRxiv : the preprint server for biology

2023

Abstract	The aryl hydrocarbon receptor (AHR) is an evolutionary conserved environmental sensor identified as indispensable regulator of epithelial homeostasis and barrier organ function. Molecular signaling cascade and target genes upon AHR activation and their contribution to cell and tissue function are however not fully understood. Multi-omics analyses using human skin keratinocytes revealed that, upon ligand activation, AHR binds open chromatin to induce expression of transcription factors (TFs), e.g., Transcription Factor AP-2α (TFAP2A), as a swift response to environmental stimuli. The terminal differentiation program including upregulation of barrier genes, filaggrin and keratins, was mediated by TFAP2A as a secondary response to AHR activation. The role of AHR-TFAP2A axis in controlling keratinocyte terminal differentiation for proper barrier formation was further confirmed using CRISPR/Cas9 in human epidermal equivalents. Overall, the study provides novel insights into the molecular mechanism behind AHR-mediated barrier function and potential novel targets for the treatment of skin barrier diseases.
Language	English
Publishing date	2023-06-08
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.06.07.544032
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Article ; Online: Investigations into the FLG Null Phenotype: Showcasing the Methodology for CRISPR/Cas9 Editing of Human Keratinocytes.

Smits, Jos P H / van den Brink, Noa J M / Meesters, Luca D / Hamdaoui, Hadia / Niehues, Hanna / Jansen, Patrick A M / van Vlijmen-Willems, Ivonne M J J / Rodijk-Olthuis, Diana / Evrard, Céline / Poumay, Yves / van Geel, Michel / Hendriks, Wiljan J A J / Schalkwijk, Joost / Zeeuwen, Patrick L J M / van den Bogaard, Ellen H

The Journal of investigative dermatology

2023 Volume 143, Issue 8, Page(s) 1520–1528.e5

Abstract: Ever since the association between FLG loss-of-function variants and ichthyosis vulgaris and atopic dermatitis disease onset was identified, FLGs function has been under investigation. Intraindividual genomic predisposition, immunological confounders, ... ...

Abstract	Ever since the association between FLG loss-of-function variants and ichthyosis vulgaris and atopic dermatitis disease onset was identified, FLGs function has been under investigation. Intraindividual genomic predisposition, immunological confounders, and environmental interactions complicate the comparison between FLG genotypes and related causal effects. Using CRISPR/Cas9, we generated human FLG-knockout (ΔFLG) N/TERT-2G keratinocytes. FLG deficiency was shown by immunohistochemistry of human epidermal equivalent cultures. Next to (partial) loss of structural proteins (involucrin, hornerin, keratin 2, and transglutaminase 1), the stratum corneum was denser and lacked the typical basket weave appearance. In addition, electrical impedance spectroscopy and transepidermal water loss analyses highlighted a compromised epidermal barrier in ΔFLG human epidermal equivalents. Correction of FLG reinstated the presence of keratohyalin granules in the stratum granulosum, FLG protein expression, and expression of the proteins mentioned earlier. The beneficial effects on stratum corneum formation were reflected by the normalization of electrical impedance spectroscopy and transepidermal water loss. This study shows the causal phenotypical and functional consequences of FLG deficiency, indicating that FLG is not only central in epidermal barrier function but also vital for epidermal differentiation by orchestrating the expression of other important epidermal proteins. These observations pave the way to fundamental investigations into the exact role of FLG in skin biology and disease.
MeSH term(s)	Humans ; CRISPR-Cas Systems ; Intermediate Filament Proteins/metabolism ; Filaggrin Proteins ; Keratinocytes/metabolism ; Phenotype
Chemical Substances	Intermediate Filament Proteins ; Filaggrin Proteins
Language	English
Publishing date	2023-03-07
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	80136-7
ISSN	1523-1747 ; 0022-202X
ISSN (online)	1523-1747
ISSN	0022-202X
DOI	10.1016/j.jid.2023.02.021
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Article ; Online: CRISPR-Cas9‒Based Genomic Engineering in Keratinocytes: From Technology to Application.

Smits, Jos P H / Meesters, Luca D / Maste, Berber G W / Zhou, Huiqing / Zeeuwen, Patrick L J M / van den Bogaard, Ellen H

JID innovations : skin science from molecules to population health

2021 Volume 2, Issue 2, Page(s) 100082

Abstract: CRISPR-Cas9 is the most straightforward genome-editing tool to date. However, its implementation across disciplines is hampered by variable genome-editing efficiencies, reduced cell viability, and low success rates in obtaining clonal cell lines. This ... ...

Abstract	CRISPR-Cas9 is the most straightforward genome-editing tool to date. However, its implementation across disciplines is hampered by variable genome-editing efficiencies, reduced cell viability, and low success rates in obtaining clonal cell lines. This review aims to recognize all CRISPR-Cas9‒related work within the experimental dermatology field to identify key factors for successful strategies in the different keratinocyte (KC) cell sources available. On the basis of these findings, we conclude that most groups use immortalized KCs for generating knockout KCs. Our critical considerations for future studies using CRISPR-Cas9, both for fundamental and clinical applications, may guide implementation strategies of CRISPR-Cas9 technologies in the (experimental) dermatology field.
Language	English
Publishing date	2021-12-01
Publishing country	Netherlands
Document type	Journal Article ; Review
ISSN	2667-0267
ISSN (online)	2667-0267
DOI	10.1016/j.xjidi.2021.100082
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