Dystrophic epidermolysis bullosa: quantitative proteomics approach to molecular mechanisms

营养不良性大疱性表皮松解症：分子机制的定量蛋白质组学方法

• 批准号: 219062131
• 负责人: Professor Dr. Jörn Dengjel
• 金额: --
• 依托单位: Départment de Biologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2019-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/219062131?language=en
• 关键词: Dystrophic; epidermolysis; bullosa; quantitative; proteomics

Dystrophic epidermolysis bullosa (DEB) is a monogenic skin disorder characterized by loss or perturbation of collagen VII functions. Patients exhibit diminished epidermal-dermal adhesion, skin blistering and subsequent scarring. Up to now, more than 800 different mutations have been disclosed in the gene COL7A1 which encodes collagen VII. A broad range of clinical phenotypes is observed, which is partially due to the nature of respective mutations and which range from isolated nail dystrophy, through localized blistering in the extremities, to generalized blistering and mutilating deformities, mucosal involvement, increased risk of skin squamous cell carcinoma and failure to thrive with premature demise. In the most severe form of the disease, due to null mutations, both collagen VII protein and anchoring fibrils are absent. In milder forms, based on missense mutations, such as glycine substitutions within the triple helical domain, collagen VII is present, but has altered stability and leads to altered functionality of anchoring fibrils. Although genotype-phenotype correlations in DEB have a certain dose-dependence, with individuals expressing partially functional collagen VII being more mildly affected than individuals completely devoid of collagen VII, it has become evident that this does not strictly apply for many individuals. This observation clearly indicates that disease manifestation in DEB is not only governed by loss of collagen VII function but importantly is also determined by secondary mechanisms. We aim to generate new knowledge of underlying molecular mechanisms of DEB by identifying hitherto unknown genes and proteins involved in disease pathogenesis and phenotypic variability. We study skin fibroblast-keratinocyte crosstalk, perturbed signal transduction and the role of inflammation in disease progression using cell culture as well as mouse models. Combination of a mouse model, cell biological and protein biochemical approaches, together with mass spectrometry-based proteomics and immunohistochemistry and -fluorescence will generate comprehensive datasets that will allow a better understanding of clinical phenotypes in DEB. Clinical phenotypes will be linked to molecular pathways, thus establishing an important prerequisite for the design of novel molecular therapies. We aim at a systems-level understanding of protein dynamics in DEB and envision that our approach will be exemplary for the investigation of other human genetic (skin) diseases.

营养不良性大疱性表皮松解症(DEB)是一种以VII型胶原功能丧失或紊乱为特征的单基因皮肤病。患者表现出表皮-真皮粘连减弱、皮肤起泡和随后的疤痕形成。到目前为止，已经在编码VII型胶原的COL7A1基因上发现了800多种不同的突变。观察到了广泛的临床表型，部分原因是各自突变的性质，范围从孤立的指甲营养不良，到四肢的局限性水泡，到全身性水泡和破坏性畸形，粘膜受累，皮肤鳞状细胞癌的风险增加，以及因过早死亡而无法茁壮成长。在最严重的疾病形式中，由于零突变，III型胶原蛋白和锚定纤维都缺失。在较温和的形式中，基于错义突变，如三螺旋结构域中的甘氨酸取代，存在II型胶原，但改变了稳定性，导致锚定纤维的功能改变。尽管DEB的基因型-表型相关性具有一定的剂量依赖性，部分表达VII型胶原的个体比完全缺乏VII型胶原的个体受到的影响更轻微，但很明显，这并不严格适用于许多个体。这一观察结果清楚地表明，DEB的疾病表现不仅与VII型胶原功能的丧失有关，更重要的是，还由次级机制决定。我们的目标是通过鉴定迄今未知的涉及疾病发病机制和表型变异的基因和蛋白，来产生对DEB潜在分子机制的新知识。我们利用细胞培养和小鼠模型研究了皮肤成纤维细胞-角质形成细胞串扰、干扰的信号转导和炎症在疾病进展中的作用。结合小鼠模型、细胞生物学和蛋白质生化方法，以及基于质谱学的蛋白质组学和免疫组织化学和荧光技术，将生成全面的数据集，从而更好地了解DEB的临床表型。临床表型将与分子通路联系起来，从而为设计新的分子疗法奠定了重要的前提。我们的目标是在系统水平上理解DEB中的蛋白质动力学，并设想我们的方法将成为研究其他人类遗传(皮肤)疾病的典范。

项目成果

Impaired lymphoid extracellular matrix impedes antibacterial immunity in epidermolysis bullosa

• DOI: 10.1073/pnas.1709111115
• 发表时间: 2018-01-23
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Nystroem, Alexander;Bornert, Olivier;Bruckner-Tuderman, Leena
• 通讯作者: Bruckner-Tuderman, Leena

Combinatorial Omics Analysis Reveals Perturbed Lysosomal Homeostasis in Collagen VII-deficient Keratinocytes*

组合组学分析揭示 VII 胶原蛋白缺乏的角质形成细胞中溶酶体稳态受到干扰*

• DOI: 10.1074/mcp.ra117.000437
• 发表时间: 2018
• 期刊: Molecular & Cellular Proteomics
• 影响因子: 7
• 作者: Thriene K;Grüning BA;Bornert O;Erxleben A;Leppert J;Athanasiou I;Weber E;Kiritsi D;Nyström A;Reinheckel T;Backofen R;Bruckner-Tuderman L;Dengjel J
• 通讯作者: Dengjel J