Inter-α-Trypsin Inhibitor Heavy Chain 5 (ITIH5) Is a Natural Stabilizer of Hyaluronan That Modulates Biological Processes in the Skin
Inter-α-Trypsin Inhibitor Heavy Chain 5 (ITIH5) Is a Natural Stabilizer of Hyaluronan That Modulates Biological Processes in the Skin
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DOI：
10.1159/000509371
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发表时间：
2020-10-01
影响因子：
2.7
通讯作者：
Baron, Jens Malte
中科院分区：
文献类型：
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作者：
Huth, Sebastian;Huth, Laura;Baron, Jens Malte
简介：透明质酸（HA）是皮肤的主要成分，发挥各种生物功能。Inter-alpha-胰蛋白酶抑制剂重链（Inter-alpha-trypsin inhibitor heavy chain，ITIH）蛋白是透明质酸（hyaladherins，HA）家族中的一个成员，其中ITIH 5在皮肤中发挥重要作用，在皮肤形态学和炎症性皮肤病（包括变应性接触性皮炎（allergic contact dermatitis，ACD））中发挥重要作用。为了研究ITIH 5在皮肤中的分子作用，我们建立了包含Itih 5基因敲除小鼠和相应野生型对照的小鼠皮肤细胞的皮肤模型。此外，建立了具有ITIH 5敲低的人真皮成纤维细胞以及鼠重组Itih 5蛋白，以使用体外粘附和HA降解测定来检查ITIH 5与HA之间的相互作用。为了更精确地理解ITIH 5在炎症性皮肤病如ACD中的作用，我们产生了KeratinoSens（R）细胞系的ITIH 5敲除细胞。结果：使用鼠皮肤模型、ITIH 5敲除成纤维细胞和活性氧（ROS）介导的HA降解测定，我们证明了ITIH 5与HA结合，从而充当HA的稳定剂。此外，微阵列分析揭示了ITIH 5对皮肤发育和ECM稳态等生物过程的影响。进行体外KeratinoSens皮肤致敏试验，我们检测到ITIH 5降低了中等和强接触敏化剂的致敏潜力。结论：两者合计，我们的实验表明，ITIH 5与HA形成复合物，从而一方面稳定HA，促进ECM结构的形成，另一方面调节炎症反应。
Introduction:Hyaluronan (HA) is a major component of the skin that exerts a variety of biological functions. Inter-alpha-trypsin inhibitor heavy chain (ITIH) proteins comprise a family of hyaladherins of which ITIH5 has recently been described in skin, where it plays a functional role in skin morphology and inflammatory skin diseases including allergic contact dermatitis (ACD).Objective:The current study focused on the ITIH5-HA interaction and its potential clinical and functional impact in extracellular matrix (ECM) stabilization.Methods:Studying the molecular effects of ITIH5 in skin, we established skin models comprising murine skin cells ofItih5knockout mice and corresponding wild-type controls. In addition, human dermal fibroblasts with an ITIH5 knockdown as well as a murine recombinantItih5protein were established to examine the interaction between ITIH5 and HA using in vitro adhesion and HA degradation assays. To understand more precisely the role of ITIH5 in inflammatory skin diseases such as ACD, we generated ITIH5 knockout cells of the KeratinoSens (R) cell line.Results:Using murine skin models, ITIH5 knockdown fibroblasts, and a reactive oxygen species (ROS)-mediated HA degradation assay, we proved that ITIH5 binds to HA, thereby acting as a stabilizer of HA. Moreover, microarray profiling revealed the impact of ITIH5 on biological processes such as skin development and ECM homeostasis. Performing the in vitro KeratinoSens skin sensitization assay, we detected that ITIH5 decreases the sensitizing potential of moderate and strong contact sensitizers.Conclusion:Taken together, our experiments revealed that ITIH5 forms complexes with HA, thereby on the one hand stabilizing HA and facilitating the formation of ECM structures and on the other hand modulating inflammatory responses.