Drummond Eleanor S, Nayak Shruti, Ueberheide Beatrix, Wisniewski Thomas
Department of Neurology, NYU Langone Medical Center, New York, NY, USA.
Proteomics Resource Center, Office of Collaborative Science, New York University School of Medicine, New York, NY, USA.
Sci Rep. 2015 Oct 21;5:15456. doi: 10.1038/srep15456.
The vast majority of human tissue specimens are formalin-fixed, paraffin embedded (FFPE) archival samples, making this type of tissue a potential gold mine for medical research. It is now accepted that proteomics can be done using FFPE tissue and can generate similar results as snap-frozen tissue. However, the current methodology requires a large amount of starting protein, limiting the questions that can be answered in these types of proteomics studies and making cell-type specific proteomics studies difficult. Cell-type specific proteomics has the potential to greatly enhance understanding of cell functioning in both normal and disease states. Therefore, here we describe a new method that allows localized proteomics on individual cell populations isolated from FFPE tissue sections using laser capture microdissection. To demonstrate this technique we microdissected neurons from archived tissue blocks of the temporal cortex from patients with Alzheimer's disease. Using this method we identified over 400 proteins in microdissected neurons; on average 78% that were neuronal and 50% that were associated with Alzheimer's disease. Therefore, this technique is able to provide accurate and meaningful data and has great potential for any future study that wishes to perform localized proteomics using very small amounts of archived FFPE tissue.
绝大多数人体组织标本是经福尔马林固定、石蜡包埋（FFPE）的存档样本，这使得这类组织成为医学研究的潜在宝库。现在人们已经认识到，可以使用FFPE组织进行蛋白质组学研究，并且能够产生与速冻组织相似的结果。然而，目前的方法需要大量起始蛋白质，这限制了这类蛋白质组学研究中能够回答的问题数量，也使得细胞类型特异性蛋白质组学研究变得困难。细胞类型特异性蛋白质组学有潜力极大地增进我们对正常和疾病状态下细胞功能的理解。因此，在这里我们描述一种新方法，该方法允许使用激光捕获显微切割技术对从FFPE组织切片中分离出的单个细胞群体进行局部蛋白质组学研究。为了演示这项技术，我们从阿尔茨海默病患者颞叶皮质的存档组织块中显微切割出神经元。使用这种方法，我们在显微切割出的神经元中鉴定出了400多种蛋白质；平均而言，78%是神经元特异性的，50%与阿尔茨海默病相关。因此，这项技术能够提供准确且有意义的数据，对于任何希望使用极少量存档FFPE组织进行局部蛋白质组学研究的未来研究都具有巨大潜力。
