A toolbox for class I HDACs reveals isoform specific roles in gene regulation and protein acetylation.
A toolbox for class I HDACs reveals isoform specific roles in gene regulation and protein acetylation.
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DOI：
10.1371/journal.pgen.1010376
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发表时间：
2022-08
期刊：
影响因子：
4.5
通讯作者：
中科院分区：
文献类型：
--
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I类组蛋白去乙酰化酶是健康和疾病中细胞命运决定的重要调节剂。虽然泛和类特异性HDAC抑制剂是可用的，但这些药物不允许全面了解个体HDAC功能，或亚型特异性靶向的治疗潜力。为了系统地比较HDAC 1、HDAC 2和HDAC 3各自催化功能的影响，我们产生了表达无催化活性HDAC酶的人HAP 1细胞系。使用这个遗传工具箱，我们比较了个体HDAC抑制与I类特异性抑制剂对细胞活力、蛋白质乙酰化和基因表达的影响。HDAC 1或HDAC 2的单独失活对细胞活力仅具有轻微影响，而HDAC 3失活或丢失导致DNA损伤和凋亡。HDAC 1/HDAC 2的失活导致COREST共阻遏物复合物组分的乙酰化增加，与该复合物相关的脱乙酰酶活性降低，神经元基因的去阻遏。HDAC 3控制核激素受体相关蛋白的乙酰化和核激素受体调节基因的表达。特异性组蛋白乙酰转移酶和HDAC的乙酰化对HDAC 1/HDAC 2的失活敏感。在广泛的测定中，我们确定特别是HDAC 1或HDAC 2催化失活模拟I类特异性HDAC抑制剂。重要的是，我们进一步证明了HDAC 1或HDAC 2的催化失活使细胞对特定的癌症药物敏感。总之，我们的系统研究揭示了HDAC 1/2/3催化功能的异构体特异性作用。我们认为，特定亚型的靶向遗传失活有效地模拟了药理学HDAC抑制，从而可以将相关HDAC鉴定为治疗干预的靶点。组蛋白脱乙酰基酶（HDAC）从组蛋白和非组蛋白蛋白去除乙酰基。乙酰化失调已被归因于各种疾病状态，包括癌症、免疫学和神经系统疾病。由于表观遗传或翻译后修饰的潜在可逆性，HDAC的药理学靶向具有很高的潜力。然而，大多数用于临床试验的HDAC抑制剂缺乏特异性，这可能导致副作用。尽管进行了多年的深入研究，但令人惊讶的是，对单个HDAC酶的底物特异性知之甚少。为了阐明个体HDAC亚型的更特异性靶向的后果，需要对每种酶的催化功能进行详细分析。尽管HDAC总体上得到了很好的研究，但由于使用不同的模型系统和频繁使用缺乏特异性的抑制剂，研究之间的交叉比较很困难。此外，常用的敲除或敲低模型消除了酶的结构功能，可能无法准确反映仅导致酶失活的抑制剂处理的情况。我们认为，使用催化失活突变体，而不是敲除可能是另一个步骤，以模仿亚型特异性酶抑制，并提供了详细的比较，并排分析I类HDAC。
The class I histone deacetylases are essential regulators of cell fate decisions in health and disease. While pan- and class-specific HDAC inhibitors are available, these drugs do not allow a comprehensive understanding of individual HDAC function, or the therapeutic potential of isoform-specific targeting. To systematically compare the impact of individual catalytic functions of HDAC1, HDAC2 and HDAC3, we generated human HAP1 cell lines expressing catalytically inactive HDAC enzymes. Using this genetic toolbox we compare the effect of individual HDAC inhibition with the effects of class I specific inhibitors on cell viability, protein acetylation and gene expression. Individual inactivation of HDAC1 or HDAC2 has only mild effects on cell viability, while HDAC3 inactivation or loss results in DNA damage and apoptosis. Inactivation of HDAC1/HDAC2 led to increased acetylation of components of the COREST co-repressor complex, reduced deacetylase activity associated with this complex and derepression of neuronal genes. HDAC3 controls the acetylation of nuclear hormone receptor associated proteins and the expression of nuclear hormone receptor regulated genes. Acetylation of specific histone acetyltransferases and HDACs is sensitive to inactivation of HDAC1/HDAC2. Over a wide range of assays, we determined that in particular HDAC1 or HDAC2 catalytic inactivation mimics class I specific HDAC inhibitors. Importantly, we further demonstrate that catalytic inactivation of HDAC1 or HDAC2 sensitizes cells to specific cancer drugs. In summary, our systematic study revealed isoform-specific roles of HDAC1/2/3 catalytic functions. We suggest that targeted genetic inactivation of particular isoforms effectively mimics pharmacological HDAC inhibition allowing the identification of relevant HDACs as targets for therapeutic intervention. Histone deacetylases (HDACs) remove acetyl groups from histones and non-histone proteins. Dysregulated acetylation has been attributed to various disease states, including cancer, immunological and neurological diseases. Due to the potential reversibility of epigenetic or post-translational modifications, the pharmacological targeting of HDACs has high potential. However, most HDAC inhibitors used for clinical trials lack specificity which might contribute to side effects. Despite many years of intense research, surprisingly little is known about the substrate specificity of individual HDAC enzymes. To elucidate the consequences of more specific targeting of individual HDAC isoforms, detailed analysis of the catalytic function of each enzyme is required. Albeit HDACs being well studied in general, cross-comparison between the studies is difficult due to the use of different model systems and the frequent use of inhibitors, which lack specificity. Also, commonly used knockout or knockdown models abolish the structural function of the enzymes, potentially not accurately reflecting the situation of inhibitor treatment which only causes enzymatic inactivation. We suggest that the use of catalytic inactive mutants instead of knockouts might be another step towards mimicking isoform specific enzyme inhibition and provide a detailed comparative, side-by-side analysis of class I HDACs.
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