Identifying Dysregulated Epigenetic Enzyme Activity in Castrate-Resistant Prostate Cancer Development.
Identifying Dysregulated Epigenetic Enzyme Activity in Castrate-Resistant Prostate Cancer Development.
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DOI：
10.1021/acschembio.6b01035
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发表时间：
2017-11-17
影响因子：
4
通讯作者：
Denu JM
中科院分区：
文献类型：
--
作者：
Lee JH;Yang B;Lindahl AJ;Damaschke N;Boersma MD;Huang W;Corey E;Jarrard DF;Denu JM
迫切需要新的策略来直接评估组蛋白修饰因子的活性，以探测与疾病进展相关的表观遗传决定因素。在这里，我们开发了一种高通量肽芯片检测方法来鉴定前列腺癌（PCa）中组蛋白赖氨酸（去乙酰化）活性的改变。这项基于微阵列的活性分析显示，与激素敏感（HS）等基因对照物相比，去势抵抗（CR） PCa细胞系中针对特定组蛋白H3位点的组蛋白乙酰转移酶（HAT）活性上调。NAD+依赖的去乙酰化实验显示，在验证的CR细胞系中sirtuin活性下调。在匹配的HS和CR细胞系之间，乙酰转移酶GCN5、PCAF、CBP和p300的水平没有变化。然而，p300在K1499位点的自乙酰化（一种已知可增强HAT活性的修饰，也是SIRT2去乙酰化的靶标）在CR细胞中高度升高，而SIRT2蛋白水平在CR细胞中降低。对HS和匹配的CR异种移植细胞系的研究显示，H3K18超乙酰化、p300活性增加和SIRT2表达降低与8/12期CR进展相关（66%）。组织芯片分析显示，H3K18的高乙酰化是CRPC的一个特征。抑制p300导致H3K18ac水平降低和雄激素受体表达增加。因此，一种新的组蛋白阵列可识别在CRPC进展过程中改变的酶活性，并可用于个性化医疗方法。SIRT2表达降低和p300活性增加导致CRPC中特定组蛋白赖氨酸位点（H3K9、H3K14和H3K18）的超乙酰化机制一致。
There is a tremendous need for novel strategies aimed at directly assessing activities of histone modifiers to probe epigenetic determinants associated with disease progression. Here, we developed a high-throughput peptide microarray assay to identify altered histone lysine (de)acetylation activity in prostate cancer (PCa). This microarray-based activity assay revealed up-regulated histone acetyltransferase (HAT) activity against specific histone H3 sites in a castrate-resistant (CR) PCa cell line compared to its hormone-sensitive (HS) isogenic counterpart. NAD+-dependent deacetylation assays revealed down-regulated sirtuin activity in validated CR lines. Levels of acetyltransferases GCN5, PCAF, CBP, and p300 were unchanged between matched HS and CR cell lines. However, autoacetylation of p300 at K1499, a modification known to enhance HAT activity and a target of deacetylation by SIRT2, was highly elevated in CR cells, while SIRT2 protein level was reduced in CR cells. Interrogation of HS and matched CR xenograft lines reveals that H3K18 hyperacetylation, increased p300 activity, and decreased SIRT2 expression are associated with progression to CR in 8/12 (66%). Tissue microarray analysis revealed that hyperacetylation of H3K18 is a feature of CRPC. Inhibition of p300 results in lower H3K18ac levels and increased expression of androgen receptors. Thus, a novel histone array identifies altered enzyme activities during the progression to CRPC and may be utilized in a personalized medicine approach. Reduced SIRT2 expression and increased p300 activity lead to a concerted mechanism of hyperacetylation at specific histone lysine sites (H3K9, H3K14, and H3K18) in CRPC.
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影响因子：
3.9
作者：
LeRoy, Gary;DiMaggio, Peter A.;Garcia, Benjamin A.
通讯作者：
Garcia, Benjamin A.
影响因子：
3.4
作者：
Britton LM;Gonzales-Cope M;Zee BM;Garcia BA
通讯作者：
Garcia BA
影响因子：
16
作者：
Black, Joshua C.;Mosley, Amber;Kitada, Tasuku;Washburn, Michael;Carey, Michael
通讯作者：
Carey, Michael
影响因子：
62.1
作者：
Huang H;Lin S;Garcia BA;Zhao Y
通讯作者：
Zhao Y
DOI：
10.1038/89668
发表时间：
2001-07-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Finnin, MS;Donigian, JR;Pavletich, NP
通讯作者：
Pavletich, NP