Anderson Kyle W, Chen Junjun, Wang Meiyao, Mast Natalia, Pikuleva Irina A, Turko Illarion V
Institute for Bioscience and Biotechnology Research, Rockville, Maryland, United States of America; Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, Maryland, United States of America; Department of Chemistry and Biochemistry, University of Maryland, College Park, Maryland, United States of America.
Institute for Bioscience and Biotechnology Research, Rockville, Maryland, United States of America; Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, Maryland, United States of America.
PLoS One. 2015 May 11;10(5):e0126592. doi: 10.1371/journal.pone.0126592. eCollection 2015.
Histone deacetylase (HDAC) inhibition has promise as a therapy for Alzheimer's disease (AD) and other neurodegenerative diseases. Currently, therapeutic HDAC inhibitors target many HDAC isoforms, a particularly detrimental approach when HDAC isoforms are known to have different and specialized functions. We have developed a multiple reaction monitoring (MRM) mass spectrometry assay using stable isotope-labeled QconCATs as internal standards to quantify HDAC isoforms. We further determined a quantitative pattern of specific HDACs expressed in various human and mouse neural tissues. In human AD frontal cortex, HDAC1,2 decreased 32%, HDAC5 increased 47%, and HDAC6 increased 31% in comparison to age-matched controls. Human neural retina concentrations of HDAC1, 2, HDAC5, HDAC6, and HDAC7 decreased in age-related macular degeneration (AMD)-affected donors and exhibited a greater decrease in AD-affected donors in comparison to age-matched control neural retinas. Additionally, HDAC concentrations were measured in whole hemisphere of brain of 5XFAD mice, a model of β-amyloid deposition, to assess similarity to AD in human frontal cortex. HDAC profiles of human frontal cortex and mouse hemisphere had noticeable differences and relatively high concentrations of HDAC3 and HDAC4 in mice, which were undetectable in humans. Our method for quantification of HDAC isoforms is a practical and efficient technique to quantify isoforms in various tissues and diseases. Changes in HDAC concentrations reported herein contribute to the understanding of the pathology of neurodegeneration.
组蛋白去乙酰化酶（HDAC）抑制有望成为治疗阿尔茨海默病（AD）和其他神经退行性疾病的方法。目前，治疗性HDAC抑制剂作用于多种HDAC亚型，然而已知HDAC亚型具有不同的特定功能，这种方法尤其有害。我们开发了一种多反应监测（MRM）质谱分析法，使用稳定同位素标记的QconCATs作为内标来定量HDAC亚型。我们进一步确定了在各种人类和小鼠神经组织中表达的特定HDAC的定量模式。与年龄匹配的对照组相比，在人类AD额叶皮质中，HDAC1、2减少了32%，HDAC5增加了47%，HDAC6增加了31%。在年龄相关性黄斑变性（AMD）患者的人神经视网膜中，HDAC1、2、HDAC5、HDAC6和HDAC7的浓度降低，与年龄匹配的对照神经视网膜相比，AD患者的降低幅度更大。此外，在β-淀粉样蛋白沉积模型5XFAD小鼠的全脑半球中测量了HDAC浓度，以评估与人类额叶皮质AD的相似性。人类额叶皮质和小鼠半球的HDAC谱有明显差异，小鼠中HDAC3和HDAC4浓度相对较高，而在人类中无法检测到。我们定量HDAC亚型的方法是一种实用且高效的技术，可用于定量各种组织和疾病中的亚型。本文报道的HDAC浓度变化有助于理解神经退行性变的病理过程。
