Development of human skin equivalents with inducible ceramide depletion for in vitro modelling of lipid deficiency

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Bibliografske podrobnosti
izdano v:bioRxiv (Jan 24, 2025)
Glavni avtor: Dina, Durotimi O
Drugi avtorji: Maiellaro, Miriam, Camera, Emanuela, Connelly, John T
Izdano:
Cold Spring Harbor Laboratory Press
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Online dostop:Citation/Abstract
Full text outside of ProQuest
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022 |a 2692-8205 
024 7 |a 10.1101/2025.01.20.633956  |2 doi 
035 |a 3159548331 
045 0 |b d20250124 
100 1 |a Dina, Durotimi O 
245 1 |a Development of human skin equivalents with inducible ceramide depletion for in vitro modelling of lipid deficiency 
260 |b Cold Spring Harbor Laboratory Press  |c Jan 24, 2025 
513 |a Working Paper 
520 3 |a The lipid composition of the epidermis plays a critical role in the barrier function of the skin, and defects in lipid synthesis or assembly can cause a spectrum of skin diseases, ranging from dry skin to severe ichthyoses. The aim of this study was to develop an in vitro model of human skin with tuneable lipid deficiency. Human N/TERT keratinocytes were engineered to express doxycycline-inducible short hairpin RNAs targeting ceramide synthase 3 (CerS3), which is essential for synthesis of ultra long chain ceramides and skin barrier function. We show that 3D human skin equivalents (HSEs) with induced knockdown of CerS3 display normal stratification and terminal differentiation but have reduced Nile Red staining for polar lipids. Further analysis of the lipidome by mass spectrometry confirmed a significant reduction in specific classes of ceramides and ceramide chain length in the CerS3 depleted HSEs. We also show that CerS3 knockdown is reversible upon removal of doxycycline and can be used to study recovery and repair of epidermal lipids. Together, these findings provide an overall strategy for genetically tuning the lipid composition within human skin models and establish a new in vitro model of ceramide deficiency.Competing Interest StatementThe authors have declared no competing interest. 
653 |a Mass spectroscopy 
653 |a Skin diseases 
653 |a Keratinocytes 
653 |a Lipid composition 
653 |a Lipids 
653 |a Doxycycline 
653 |a Epidermis 
653 |a Ceramide 
700 1 |a Maiellaro, Miriam 
700 1 |a Camera, Emanuela 
700 1 |a Connelly, John T 
773 0 |t bioRxiv  |g (Jan 24, 2025) 
786 0 |d ProQuest  |t Biological Science Database 
856 4 1 |3 Citation/Abstract  |u https://www.proquest.com/docview/3159548331/abstract/embedded/75I98GEZK8WCJMPQ?source=fedsrch 
856 4 0 |3 Full text outside of ProQuest  |u https://www.biorxiv.org/content/10.1101/2025.01.20.633956v1