Altered Protein Function Caused by AMD-associated Variant rs704 Links Vitronectin to Disease Pathology.
Altered Protein Function Caused by AMD-associated Variant rs704 Links Vitronectin to Disease Pathology.
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DOI：
10.1167/iovs.61.14.2
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发表时间：
2020-12-01
影响因子：
4.4
通讯作者：
Friedrich U
中科院分区：
文献类型：
--
作者：
Biasella F;Plössl K;Karl C;Weber BHF;Friedrich U
Vitronectin是一种细胞黏附和扩散因子，被认为在老年性黄斑变性(AMD)的发病机制中发挥了作用，因为它是AMD特异性细胞外沉积(例如软性玻璃体、视网膜下玻璃体沉积)的主要成分。本研究探讨了玻璃连接蛋白编码基因VTN中与AMD相关的非同义变异rs704对玻璃连接蛋白功能的影响。通过人诱导多能干细胞(HiPSCs)和人神经视网膜来源的视网膜色素上皮(RPE)细胞VTN转录的半定量测序以及异源表达的vitronectin异构体的蛋白质印迹分析，分析了rs704对Vitronectin表达和加工的影响。免疫印迹法检测玻璃连结蛋白亚型与视网膜和内皮细胞的结合。免疫荧光染色后细胞外基质(ECM)沉积在培养的RPE细胞中，异源表达Vitronectin异构体。荧光标记的RPE或内皮细胞对重组玻璃连素或含玻璃连素的ECM的粘附性进行了荧光或显微镜下的研究。管形成和迁移分析研究了Vitronectin对血管生成相关过程的影响。突变体rs704影响Vitronectin的表达、分泌和加工，但不影响寡聚化。AMD风险相关蛋白亚型和非AMD风险相关蛋白亚型之间的细胞结合和对RPE介导的ECM沉积的影响不同。最后，Vitronectin影响黏附和内皮管的形成。AMD风险相关的Vitronectin亚型在与AMD病理的亚RPE相关的细胞过程中表现出表达增加和功能改变。虽然需要进一步的研究来解决视网膜下疾病方面的问题，但这项初步研究支持玻璃连结蛋白参与AMD的发病机制。
Vitronectin, a cell adhesion and spreading factor, is suspected to play a role in the pathogenesis of age-related macular degeneration (AMD), as it is a major component of AMD-specific extracellular deposits (e.g., soft drusen, subretinal drusenoid deposits). The present study addressed the impact of AMD-associated non-synonymous variant rs704 in the vitronectin-encoding gene VTN on vitronectin functionality. Effects of rs704 on vitronectin expression and processing were analyzed by semi-quantitative sequencing of VTN transcripts from retinal pigment epithelium (RPE) cells generated from human induced pluripotent stem cells (hiPSCs) and from human neural retina, as well as by western blot analyses on heterologously expressed vitronectin isoforms. Binding of vitronectin isoforms to retinal and endothelial cells was analyzed by western blot. Immunofluorescence staining followed extracellular matrix (ECM) deposition in cultured RPE cells heterologously expressing the vitronectin isoforms. Adhesion of fluorescently labeled RPE or endothelial cells in dependence of recombinant vitronectin or vitronectin-containing ECM was investigated fluorometrically or microscopically. Tube formation and migration assays addressed effects of vitronectin on angiogenesis-related processes. Variant rs704 affected expression, secretion, and processing but not oligomerization of vitronectin. Cell binding and influence on RPE-mediated ECM deposition differed between AMD-risk-associated and non-AMD-risk-associated protein isoforms. Finally, vitronectin affected adhesion and endothelial tube formation. The AMD-risk-associated vitronectin isoform exhibits increased expression and altered functionality in cellular processes related to the sub-RPE aspects of AMD pathology. Although further research is required to address the subretinal disease aspects, this initial study supports an involvement of vitronectin in AMD pathogenesis.
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影响因子：
3.5
作者：
Brandl, Caroline;Zimmermann, Stephanie J.;Milenkovic, Vladimir M.;Rosendahl, Sibylle M. G.;Grassmann, Felix;Milenkovic, Andrea;Hehr, Ute;Federlin, Marianne;Wetzel, Christian H.;Helbig, Horst;Weber, Bernhard H. F.
通讯作者：
Weber, Bernhard H. F.
影响因子：
4.4
作者：
Ablonczy, Zsolt;Dahrouj, Mohammad;Crosson, Craig E.
通讯作者：
Crosson, Craig E.
影响因子：
56.9
作者：
BROOKS, PC;CLARK, RAF;CHERESH, DA
通讯作者：
CHERESH, DA
影响因子：
2.6
作者：
Date, Kimie;Suzuki, Risa;Ogawa, Haruko
通讯作者：
Ogawa, Haruko
影响因子：
17.8
作者：
Datta S;Cano M;Ebrahimi K;Wang L;Handa JT
通讯作者：
Handa JT