Transthyretin affects the proliferation and migration of human retinal microvascular endothelial cells in hyperglycemia via hnRNPA2B1
Transthyretin affects the proliferation and migration of human retinal microvascular endothelial cells in hyperglycemia via hnRNPA2B1
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DOI：
10.1016/j.bbrc.2021.04.035
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发表时间：
2021-04-21
影响因子：
3.1
通讯作者：
Shao, Jun
中科院分区：
文献类型：
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作者：
Gu, Yu;Hu, Di;Shao, Jun
甲状腺素运载蛋白（Transthyretin，TTR）通过调控STAT-4/miR-223- 3 p/FBXW 7信号通路中及其下游的分子，抑制糖尿病视网膜病变环境中的新生血管形成，但其直接靶点尚不清楚。用免疫共沉淀（co-IP）和质谱（MS）技术筛选了TTR与人视网膜微血管内皮细胞（hRECs）核内靶点hnRNP A2 B1的相互作用，并进一步证实了TTR与hRECs的相互作用。关于使用Discovery Studio进行的ZDOCK分析，模拟了TTR和hnRNPA 2B 1之间的界面和潜在结合位点;在这些区域设计了突变体，并重组表达和制备了5个可溶性突变体; TTR和hnRNPA 2B 1之间的相互作用被几个突变残基破坏。此外，对于几种突变的TTR，对hREC的增殖，迁移和管形成的抑制活性在体外是不存在的。TTR-hnRNPA 2B 1被破坏后，STAT-4/miR-223- 3 p/FBXW 7信号通路中及其下游的分子，包括STAT-4、miR-223- 3 p、FBXW 7 p-Akt和Notch 1均不能被TTR突变体调控，因此提出了一个TTR-hnRNPA 2B 1/STAT-4/miR-223- 3 p/FBXW 7。本研究提示TTR可能通过与hnRNPA 2B 1直接结合而在糖尿病环境中发挥生理作用，为临床诊断、治疗及进一步应用提供了理论依据。(c)2021爱思唯尔公司All rights reserved.
Transthyretin (TTR) has been proved to repress neovascularization in diabetic retinopathy environment by regulating the molecules in and downstream of the STAT-4/miR-223-3p/FBXW7 signal pathway; however, the details of its direct targets are still not well understood. The interaction between TTR and a target in nucleus of human retinal microvascular endothelial cells (hRECs), heterogeneous nuclear ribonucleoprotein (hnRNP) A2B1, was screened by immunoprecipitation (IP) and mass spectrum (MS), and it was further confirmed by co-immunoprecipitation (co-IP). Regarding ZDOCK analysis using Discovery Studio, the interface and potential binding sites between TTR and hnRNPA2B1 were simulated; mutants were designed in these regions and five soluble ones were recombinantly expressed and prepared; the interaction between TTR and hnRNPA2B1 were disrupted by several mutated residues. In addition, for several mutated TTRs, the inhibition activities against the proliferation, migration and tube formation of hRECs were absent in vitro. Following the disruption of TTR-hnRNPA2B1, the molecules in and downstream of STAT-4/miR-223-3p/FBXW7 signal pathway, including STAT-4, miR-223-3p, FBXW7 p-Akt and Notch1 could not be regulated by TTR mutants; therefore, a TTR-hnRNPA2B1/STAT-4/miR-223-3p/FBXW7 was proposed. In conclusion, this work suggested that TTR should play a physiological role in diabetic environment by the direct binding with hnRNPA2B1, and it provided a theoretical basis for clinical diagnosis, therapy and further application.(c) 2021 Elsevier Inc. All rights reserved.