Kelly John J, Esseltine Jessica L, Shao Qing, Jabs Ethylin Wang, Sampson Jacinda, Auranen Mari, Bai Donglin, Laird Dale W
Anatomy and Cell Biology, University of Western Ontario, London, ON N6A 5C1, Canada.
Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029 Johns Hopkins University, Baltimore, MD 21205.
Mol Biol Cell. 2016 Jul 15;27(14):2172-85. doi: 10.1091/mbc.E16-01-0062. Epub 2016 May 25.
Oculodentodigital dysplasia (ODDD) is a rare genetic disease that affects the development of multiple organs in the human body. More than 70 mutations in the gap junction connexin43 (Cx43) gene, GJA1, are associated with ODDD, most of which are inherited in an autosomal dominant manner. Many patients exhibit similar clinical presentations. However, there is high intrafamilial and interfamilial phenotypic variability. To better understand this variability, we established primary human dermal fibroblast cultures from several ODDD patients and unaffected controls. In the present study, we characterized three fibroblast lines expressing heterozygous p.L7V, p.G138R, and p.G143S Cx43 variants. All ODDD fibroblasts exhibited slower growth, reduced migration, and defective cell polarization, traits common to all ODDD fibroblasts studied so far. However, we found striking differences in overall expression levels, with p.L7V down-regulated at the mRNA and protein level. Although all of the Cx43 variants could traffic to the cell surface, there were stark differences in gap junction plaque formation, gap junctional intercellular communication, Cx43 phosphorylation, and hemichannel activity among Cx43 variants, as well as subtle differences in myofibroblast differentiation. Together these findings enabled us to discover mutation-specific pathologies that may help to predict future clinical outcomes.
眼牙指发育不全(ODDD)是一种罕见的遗传性疾病,会影响人体多个器官的发育。缝隙连接蛋白43(Cx43)基因GJA1中的70多种突变与ODDD相关,其中大多数以常染色体显性方式遗传。许多患者表现出相似的临床表现。然而,家族内和家族间存在高度的表型变异性。为了更好地理解这种变异性,我们从几名ODDD患者和未受影响的对照中建立了原代人皮肤成纤维细胞培养物。在本研究中,我们对表达杂合p.L7V、p.G138R和p.G143S Cx43变体的三种成纤维细胞系进行了表征。所有ODDD成纤维细胞均表现出生长缓慢、迁移减少和细胞极化缺陷,这些特征是迄今为止所有研究的ODDD成纤维细胞共有的。然而,我们发现总体表达水平存在显著差异,p.L7V在mRNA和蛋白质水平均下调。尽管所有Cx43变体都可以转运到细胞表面,但Cx43变体之间在缝隙连接斑块形成、缝隙连接细胞间通讯、Cx43磷酸化和半通道活性方面存在明显差异,在肌成纤维细胞分化方面也存在细微差异。这些发现共同使我们能够发现特定于突变的病理情况,这可能有助于预测未来的临床结果。