Crosstalk Between Histone and DNA Methylation in Regulation of Retinal Matrix Metalloproteinase-9 in Diabetes.
Crosstalk Between Histone and DNA Methylation in Regulation of Retinal Matrix Metalloproteinase-9 in Diabetes.
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DOI：
10.1167/iovs.17-22706
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发表时间：
2017-12-01
影响因子：
4.4
通讯作者：
Kowluru RA
中科院分区：
文献类型：
--
作者：
Duraisamy AJ;Mishra M;Kowluru RA
糖尿病激活基质金属蛋白酶-9 (matrix metalloproteinase-9, MMP-9)， MMP-9通过损伤视网膜线粒体激活毛细血管细胞凋亡。MMP-9启动子具有许多转录因子的结合位点，在糖尿病中其启动子经历表观遗传修饰，包括组蛋白修饰和DNA甲基化。Zeste同源物2 （Ezh2）的增强子，催化组蛋白3赖氨酸27 （H3K27me2和me3）的二甲基化/三甲基化，也与DNA甲基化有关。我们的目的是研究组蛋白和DNA修饰在MMP-9调控中的联系。利用人视网膜内皮细胞和糖尿病大鼠视网膜微血管，采用染色质免疫沉淀技术定量高血糖对H3K27me3的影响，以及MMP-9启动子上Ezh2的募集。H3K27三甲基化在调节MMP-9 DNA甲基化转录中的作用是通过Ezh2的特异性siRNA和药物抑制剂来确定的。高血糖升高H3K27me3水平和Ezh2在MMP-9启动子处的募集，并增加Ezh2的酶活性。Ezh2的抑制减弱了DNA甲基化（Dnmt1）和羟甲基化（Tet2）酶以及5羟甲基胞嘧啶在MMP-9启动子同一区域的募集，并阻止了MMP-9转录的增加和线粒体损伤。糖尿病患者通过H3K27的三甲基化激活Ezh2，促进了负责调节MMP-9启动子DNA甲基化的酶的募集，从而导致其转录激活。因此，糖尿病中H3K27三甲基化和DNA甲基化之间的密切串扰在维持MMP-9的细胞表观遗传完整性中起着关键作用。
Diabetes activates matrix metalloproteinase-9 (MMP-9), and MMP-9 via damaging retinal mitochondria, activates capillary cell apoptosis. MMP-9 promoter has binding sites for many transcription factors, and in diabetes its promoter undergoes epigenetic modifications, including histone modifications and DNA methylation. Enhancer of Zeste homolog 2 (Ezh2), which catalyzes dimethylation/trimethylation of histone 3 lysine 27 (H3K27me2 and me3), is also associated with DNA methylation. Our aim was to investigate link(s) between histone and DNA modifications in the regulation of MMP-9. Using human retinal endothelial cells, and also retinal microvessels from diabetic rats, effect of hyperglycemia on H3K27me3, and recruitment of Ezh2 at the MMP-9 promoter were quantified by chromatin-immunoprecipitation technique. Role of H3K27 trimethylation in regulating DNA methylation-transcription of MMP-9 was determined by regulating Ezh2 by its specific siRNA and also a pharmacologic inhibitor. Hyperglycemia elevated H3K27me3 levels and the recruitment of Ezh2 at the MMP-9 promoter, and increased the enzyme activity of Ezh2. Inhibition of Ezh2 attenuated recruitment of both DNA methylating (Dnmt1) and hydroxymethylating (Tet2) enzymes and 5 hydroxymethyl cytosine at the same region of the MMP-9 promoter, and prevented increase in MMP-9 transcription and mitochondrial damage. Activation of Ezh2 in diabetes, via trimethylation of H3K27, facilitates recruitment of the enzymes responsible for regulation of DNA methylation of the MMP-9 promoter, resulting in its transcriptional activation. Thus, a close crosstalk between H3K27 trimethylation and DNA methylation in diabetes plays a critical role in the maintenance of cellular epigenetic integrity of MMP-9.
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DOI：
10.1038/labinvest.2010.89
发表时间：
2010-09
期刊：
Laboratory investigation; a journal of technical methods and pathology
影响因子：
--
作者：
通讯作者：
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影响因子：
15.9
作者：
Mizutani, M;Kern, TS;Lorenzi, M
通讯作者：
Lorenzi, M
影响因子：
15.9
作者：
Barber, AJ;Lieth, E;Gardner, TW
通讯作者：
Gardner, TW
影响因子：
10.5
作者：
Fischle, Wolfgang
通讯作者：
Fischle, Wolfgang
影响因子：
4.4
作者：
Mohammad, Ghulam;Kowluru, Renu A.
通讯作者：
Kowluru, Renu A.