Epidermolysis bullosa simplex due to KRT5 mutations: mutation-related differences in cellular fragility and the protective effects of trimethylamine N-oxide in cultured primary keratinocytes
Epidermolysis bullosa simplex due to KRT5 mutations: mutation-related differences in cellular fragility and the protective effects of trimethylamine N-oxide in cultured primary keratinocytes
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DOI：
10.1111/j.1365-2133.2009.09615.x
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发表时间：
2010-05-01
影响因子：
10.3
通讯作者：
Torma, H.
中科院分区：
文献类型：
--
作者：
Chamcheu, J. C.;Virtanen, M.;Torma, H.
单纯大疱性表皮病（epiderbullosa simplex，EBS）是一种以基底角蛋白细胞溶解和表皮内水疱为特征的机械性大疱性皮肤脆性疾病，常由角蛋白基因（KRT 5或KRT 14）突变引起。不存在补救措施，这些疾病提出了一个需要开发的新therapy.ObjectivesTo确定新的基因型-表型的关系，在体内和体外培养的原代EBS角质形成细胞，并研究细胞骨架稳定作用的三甲胺N-氧化物（TMAO）在热应激的EBS cells.MethodsGenomic DNA和cDNA样本从三个瑞典的EBS患者角蛋白突变进行了分析。主要EBS角质形成细胞培养物的建立，热应激和不加TMAO，随后通过细胞fragmental.ResultsIn除了以前报道的KRT 5突变（V186 L）在一个病人，两名患者被发现有一个新的I183 M和复发E475 G更换KRT 5的评价。培养的EBS角质形成细胞没有表现出角蛋白聚集体或细胞损失，除了在p.I183M突变的患者中显示3%的聚集体和2%的细胞损失。在瞬时热应激后，在p.I183M、p.E475G和p.V186L突变体细胞中，含聚集体的细胞数量分别增加至21%、27%和13%。有趣的是，预处理TMAO热应激前，剂量依赖性地减少了聚集的细胞和细胞loss.ConclusionThese结果揭示了基因型-表型相关性在EBS角质形成细胞热应激后，并建议蛋白质稳定作为一种新的治疗策略。
P>BackgroundEpidermolysis bullosa simplex (EBS) is a mechanobullous skin fragility disease characterized by cytolysis of basal keratinocytes and intraepidermal blistering often caused by mutations in keratin genes (KRT5 or KRT14). No remedies exist for these disorders presenting a need for development of novel therapies.ObjectivesTo identify new genotype-phenotype relationships in vivo and in cultured primary EBS keratinocytes in vitro, and to study the cytoskeletal stabilizing effects of trimethylamine N-oxide (TMAO) in heat-stressed EBS cells.MethodsGenomic DNA and cDNA samples from three Swedish patients with EBS were analysed for keratin mutations. Primary EBS keratinocyte cultures were established, heat stressed with and without added TMAO, followed by evaluation of cellular fragility.ResultsIn addition to the previously reported KRT5 mutation (V186L) in one patient, two patients were found to have a novel I183M and recurrent E475G replacements in KRT5. Cultured EBS keratinocytes did not exhibit keratin aggregates or cell loss, except in the patient with the p.I183M mutation who showed 3% aggregates and 2% cell loss. Upon transient heat stress the number of aggregate-containing cells increased to 21%, 27% and 13%, respectively, in the p.I183M, p.E475G and p.V186L mutant cells. Interestingly, pretreatment with TMAO prior to heat stress, dose dependently reduced the number of aggregate-containing cells and cell loss.ConclusionThese results revealed a genotype-phenotype correlation in EBS keratinocytes upon heat stress and suggest protein stabilization as a new therapeutic strategy.