Deletion of TNFAIP6 gene in human keratinocytes demonstrates a role for TSG-6 to retain hyaluronan inside epidermis

Data collected about TSG-6-/- human reconstructed epidermis (RHE): TSG-6 is an anti-inflammatory protein which interacts with hyaluronan (HA) in extracellular matrix between keratinocytes but its function is not well understood. As TSG-6 is more expressed under pathological conditions, it was studied using RHE stimulated with Th2 cytokines or RHE infected by dermatophytes. In both two inflammatory models, TSG-6 and HA are significantly overexpressed. In order to understand the role of TSG-6, immortalized N/TERT keratinocytes were edited using CRISPR/Cas9 and two TSG-6-/- clonal populations were created and compared to TSG-6+/+ keratinocytes. TSG-6-/- keratinocytes showed a migration ability slowdown relative to TSG-6+/+ cells. Moreover, even if TSG-6-/- RHE exhibit normal differentiation, morphology and transcriptome, an increased HA leakage in the underlying culture medium is measured, especially in inflammatory conditions. This drastic HA leakage is rescued when TSG-6 expressing keratinocytes are reintroduced in RHE. Results suggest an implication of TSG-6 in HA remodelling and retaining in epidermal extracellular matrix.

关于TSG-6-/-人类重构表皮（RHE）的数据收集：TSG-6是一种抗炎蛋白，与角质形成细胞之间的细胞外基质中的透明质酸（HA）相互作用，但其功能尚不清楚。由于TSG-6在病理条件下表达更为显著，因此利用受Th2细胞因子刺激的RHE或受皮肤癣菌感染的RHE进行了研究。在这两种炎症模型中，TSG-6和HA均显著过表达。为了理解TSG-6的作用，利用CRISPR/Cas9技术对N/TERT角质形成细胞进行编辑，创建了两个TSG-6-/-克隆种群，并与TSG-6+/+角质形成细胞进行比较。与TSG-6+/+细胞相比，TSG-6-/-角质形成细胞的迁移能力有所下降。此外，尽管TSG-6-/- RHE表现出正常的分化、形态和转录组，但在基础培养基中的HA泄漏量增加，尤其是在炎症条件下。当将表达TSG-6的角质形成细胞重新引入RHE时，这种剧烈的HA泄漏得到缓解。结果表明，TSG-6在HA重塑和表皮细胞外基质中的保留过程中具有重要作用。