451 Modeling darier disease using gene-edited human keratinocytes and organotypic epidermis to identify therapeutic targets
نویسندگان
چکیده
Darier disease(DD) is a genetic disorder characterized by impaired intercellular adhesion and aberrant differentiation of keratinocytes, leading to hyperkeratotic papules epidermal erosions. While mutations underlying DD are known target SERCA2, an endoplasmic reticulum(ER) calcium pump, current therapies for this chronic, painful skin limited some, such as oral retinoids, may pose significant long-term risks. Unfortunately, mice lacking SERCA2 do not replicate the cutaneous findings seen in human patients. Thus, we aimed establish vitro model improve our understanding its pathogenesis downstream deficiency reveal new potential therapeutic strategies. TERT-immortalized keratinocytes were subjected CRISPR/Cas9 gene editing generate cells deficient SERCA2. Transduction with GCaMP, cytosolic sensor, allowed us demonstrate augmented spikes intracellular SERCA2-depleted upon exposure elevated extracellular calcium. Culturing SERCA2-deficient organotypic epidermis revealed features histopathology including dyskeratosis reduced adhesion. To elucidate pathogenic drivers resulting from deficiency, performed RNA sequencing heterozygous or homozygous loss dysregulated homeostasis due depletion expected disrupt numerous cellular pathways, preliminary analysis expression profiles alterations potentially druggable targets, regulators cornification, adhesion, homeostasis, inflammatory responses, cell death. Further studies ongoing assess whether dampening effects using existing pharmacologic compounds can effectively reverse pathology epidermis.
منابع مشابه
C/EBPδ Gene Targets in Human Keratinocytes
C/EBPs are a family of B-Zip transcription factors--TFs--involved in the regulation of differentiation in several tissues. The two most studied members--C/EBPα and C/EBPβ--play important roles in skin homeostasis and their ablation reveals cells with stem cells signatures. Much less is known about C/EBPδ which is highly expressed in the granular layer of interfollicular epidermis and is a direc...
متن کاملUsing proteomics to identify potential therapeutic targets in platelets.
Proteomics has provided powerful new insights into the complex events of the anucleate platelet and has revealed many potential protein targets in the search for suitable agents for thrombotic disease. In the present study, we summarize recent proteomic approaches to analyse specific platelet subproteomes, such as the platelet releasate, the platelet phosphotyrosine proteome and characterizatio...
متن کاملRegulation of ABCA1 expression in human keratinocytes and murine epidermis
Keratinocytes require abundant cholesterol for cutaneous permeability barrier function; hence, the regulation of cholesterol homeostasis is of great importance. ABCA1 is a membrane transporter responsible for cholesterol efflux and plays a pivotal role in regulating cellular cholesterol levels. We demonstrate that ABCA1 is expressed in cultured human keratinocytes (CHKs) and murine epidermis. L...
متن کاملRegulation of ABCG1 expression in human keratinocytes and murine epidermis.
ABCG1, a member of the ATP binding cassette superfamily, facilitates the efflux of cholesterol from cells to HDL. In this study, we demonstrate that ABCG1 is expressed in cultured human keratinocytes and murine epidermis, and induced during keratinocyte differentiation, with increased levels in the outer epidermis. ABCG1 is regulated by liver X receptor (LXR) and peroxisome proliferator-activat...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Journal of Investigative Dermatology
سال: 2022
ISSN: ['1523-1747', '0022-202X']
DOI: https://doi.org/10.1016/j.jid.2022.05.460