G-quadruplexes Stabilization Upregulates CCN1 and Accelerates Aging in Cultured Cerebral Endothelial Cells.
G-quadruplexes Stabilization Upregulates CCN1 and Accelerates Aging in Cultured Cerebral Endothelial Cells.
复制标题
DOI：
10.3389/fragi.2021.797562
复制
发表时间：
2021
影响因子：
--
通讯作者：
Moruno-Manchon, Jose Felix
中科院分区：
文献类型：
--
作者：
Noh, Brian;Blasco-Conesa, Maria P.;Lai, Yun-Ju;Ganesh, Bhanu Priya;Urayama, Akihiko;Moreno-Gonzalez, Ines;Marrelli, Sean P.;Mccullough, Louise D.;Moruno-Manchon, Jose Felix
脑内皮细胞的衰老被认为是导致脑血管功能障碍的一种机制，而脑血管功能障碍是血管性痴呆的先兆。富含半胱氨酸的血管生成诱导因子61（Cyr 61/CCN 1）是由脑内皮细胞（CEC）分泌的一种基质细胞蛋白。CCN 1诱导成纤维细胞衰老。然而，CCN 1是否有助于CEC的衰老以及如何调节需要进一步研究。衰老与DNA和RNA富G基序中四链鸟嘌呤四链体（G4 s）的形成有关。G4结构的稳定化根据基因靶通过上调或下调来调节转录和翻译。以前，我们发现用G4稳定化合物治疗的老年小鼠在其大脑中具有增强的衰老相关（SA）表型，并且这些小鼠表现出增强的认知缺陷。人CCN 1 mRNA的3′-UTR中的一个序列具有在体外折叠成G4的能力。我们假设G4稳定化调节培养的原代CEC中的CCN 1并诱导内皮衰老。我们使用脑微血管组分和培养的初级CEC从年轻（4个月大，m/o）和老年（18-m/o）小鼠，以确定CCN 1水平。SA表型通过高分辨率荧光显微镜在培养的原代CEC中确定，并且我们使用硫磺素T识别RNA-G4 s用于荧光光谱。我们发现，从老年小鼠培养的CEC表现出增强水平的SA表型，和更高水平的CCN 1和G4稳定。在培养的CEC中，CCN 1诱导SA表型，如SA β-半乳糖苷酶活性和双链DNA损伤。此外，CCN 1水平被G4配体上调，并且Ccn 1 mRNA的3′-UTR中的富含G的基序被折叠成G4。总之，我们表明，CCN 1可以诱导培养的初级CEC衰老，我们提供的证据表明，G4稳定是一种新的机制，调节SASP组件CCN 1。
Senescence in the cerebral endothelium has been proposed as a mechanism that can drive dysfunction of the cerebral vasculature, which precedes vascular dementia. Cysteine-rich angiogenic inducer 61 (Cyr61/CCN1) is a matricellular protein secreted by cerebral endothelial cells (CEC). CCN1 induces senescence in fibroblasts. However, whether CCN1 contributes to senescence in CEC and how this is regulated requires further study. Aging has been associated with the formation of four-stranded Guanine-quadruplexes (G4s) in G-rich motifs of DNA and RNA. Stabilization of the G4 structures regulates transcription and translation either by upregulation or downregulation depending on the gene target. Previously, we showed that aged mice treated with a G4-stabilizing compound had enhanced senescence-associated (SA) phenotypes in their brains, and these mice exhibited enhanced cognitive deficits. A sequence in the 3′-UTR of the human CCN1 mRNA has the ability to fold into G4s in vitro. We hypothesize that G4 stabilization regulates CCN1 in cultured primary CEC and induces endothelial senescence. We used cerebral microvessel fractions and cultured primary CEC from young (4-months old, m/o) and aged (18-m/o) mice to determine CCN1 levels. SA phenotypes were determined by high-resolution fluorescence microscopy in cultured primary CEC, and we used Thioflavin T to recognize RNA-G4s for fluorescence spectra. We found that cultured CEC from aged mice exhibited enhanced levels of SA phenotypes, and higher levels of CCN1 and G4 stabilization. In cultured CEC, CCN1 induced SA phenotypes, such as SA β-galactosidase activity, and double-strand DNA damage. Furthermore, CCN1 levels were upregulated by a G4 ligand, and a G-rich motif in the 3′-UTR of the Ccn1 mRNA was folded into a G4. In conclusion, we demonstrate that CCN1 can induce senescence in cultured primary CEC, and we provide evidence that G4 stabilization is a novel mechanism regulating the SASP component CCN1.
登录
查看更多内容
影响因子：
14.9
作者：
Renaud de la Faverie A;Guédin A;Bedrat A;Yatsunyk LA;Mergny JL
通讯作者：
Mergny JL
影响因子：
4.5
作者：
Aldahmash A
通讯作者：
Aldahmash A
DOI：
10.15252/embj.201488663
发表时间：
2014-11-03
期刊：
The EMBO journal
影响因子：
--
作者：
Castillo Bosch P;Segura-Bayona S;Koole W;van Heteren JT;Dewar JM;Tijsterman M;Knipscheer P
通讯作者：
Knipscheer P
DOI：
10.1073/pnas.92.20.9363
发表时间：
1995-09-26
影响因子：
11.1
作者：
DIMRI, GP;LEE, XH;CAMPISI, J
通讯作者：
CAMPISI, J
影响因子：
14.9
作者：
Guo K;Gokhale V;Hurley LH;Sun D
通讯作者：
Sun D