title: Knock-down of filaggrin does not affect lipid organization and composition in stratum corneum of reconstructed human skin equivalents.
abstract: Human skin mainly functions as an effective barrier against unwanted environmental influences. The barrier function strongly relies on the outermost layer of the skin, the stratum corneum (SC), which is composed of corneocytes embedded in an extracellular lipid matrix. The importance of a proper barrier function is shown in various skin disorders such as atopic dermatitis (AD), a complex human skin disorder strongly associated with filaggrin (FLG) null mutations, but their role in barrier function is yet unclear. To study the role of FLG in SC barrier properties in terms of SC lipid organization and lipid composition, we generated an N/TERT-based 3D-skin equivalent (NSE) after knock-down of FLG with shRNA. In these NSEs, we examined epidermal morphogenesis by evaluating the expression of differentiation markers keratin 10, FLG, loricrin and the proliferation marker ki67. Furthermore, the SC was extensively analysed for lipid organization, lipid composition and SC permeability. Our results demonstrate that FLG knock-down (FLG-KD) did not affect epidermal morphogenesis, SC lipid organization, lipid composition and SC permeability for a lipophilic compound in NSEs. Therefore, our findings indicate that FLG-KD alone does not necessarily affect the functionality of a proper barrier function.
authors.affiliation: Gorlaeus Laboratories, Department of Drug Delivery Technology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, the Netherlands; Department of Dermatology, Leiden University Medical Centre, Leiden, the Netherlands.
authors.forename: Vincent
authors.initials: V
authors.lastname: van Drongelen
authors.forename: Mariam
authors.initials: M
authors.lastname: Alloul-Ramdhani
authors.forename: Mogbekeloluwa O
authors.initials: MO
authors.lastname: Danso
authors.forename: Arnout
authors.initials: A
authors.lastname: Mieremet
authors.forename: Aat
authors.initials: A
authors.lastname: Mulder
authors.forename: Jeroen
authors.initials: J
authors.lastname: van Smeden
authors.forename: Joke A
authors.initials: JA
authors.lastname: Bouwstra
authors.forename: Abdoelwaheb
authors.initials: A
authors.lastname: El Ghalbzouri
mesh_terms: D049109:Cell Proliferation; D003876:Dermatitis, Atopic / Q000473:pathology; D004817:Epidermis / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D000091344:Filaggrin Proteins; D055785:Gene Knockdown Techniques; D006579:Heterozygote; D006801:Humans; D007249:Inflammation; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism*; D053550:Keratin-10 / Q000378:metabolism; D007633:Keratins / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000378:metabolism; D010539:Permeability; D010641:Phenotype; D012867:Skin / Q000473:pathology*; D012871:Skin Diseases / Q000473:pathology
chemical_list: C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D019394:Ki-67 Antigen; D008055:Lipids; D008565:Membrane Proteins; C064554:loricrin; D053550:Keratin-10; D007633:Keratins
keywords: N/TERT; filaggrin; skin barrier; skin equivalent; stratum corneum