Sadick Jessica S, Crawford Lorin A, Cramer Harry C, Franck Christian, Liddelow Shane A, Darling Eric M
Department of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, RI, 02912, USA.
Neuroscience Institute, NYU Langone Medical Center, New York, NY, 10016, USA.
Ann Biomed Eng. 2020 Aug;48(8):2218-2232. doi: 10.1007/s10439-020-02507-y. Epub 2020 Apr 17.
Here we demonstrate a technique to generate proteomic signatures of specific cell types within heterogeneous populations. While our method is broadly applicable across biological systems, we have limited the current work to study neural cell types isolated from human, post-mortem Alzheimer's disease (AD) and aged-matched non-symptomatic (NS) brains. Motivating the need for this tool, we conducted an initial meta-analysis of current, human AD proteomics studies. While the results broadly corroborated major neurodegenerative disease hypotheses, cell type-specific predictions were limited. By adapting our Formaldehyde-fixed Intracellular Target-Sorted Antigen Retrieval (FITSAR) method for proteomics and applying this technique to characterize AD and NS brains, we generated enriched neuron and astrocyte proteomic profiles for a sample set of donors (available at www.fitsarpro.appspot.com ). Results showed the feasibility for using FITSAR to evaluate cell-type specific hypotheses. Our overall methodological approach provides an accessible platform to determine protein presence in specific cell types and emphasizes the need for protein-compatible techniques to resolve systems complicated by cellular heterogeneity.
在此，我们展示了一种在异质群体中生成特定细胞类型蛋白质组特征的技术。虽然我们的方法广泛适用于各种生物系统，但目前我们将工作局限于研究从人类尸检后的阿尔茨海默病（AD）和年龄匹配的无症状（NS）大脑中分离出的神经细胞类型。出于对这种工具的需求，我们对当前的人类AD蛋白质组学研究进行了初步的荟萃分析。虽然结果大致证实了主要的神经退行性疾病假说，但细胞类型特异性的预测有限。通过将我们的甲醛固定细胞内靶点分选抗原修复（FITSAR）方法应用于蛋白质组学，并将该技术应用于表征AD和NS大脑，我们为一组供体生成了富集的神经元和星形胶质细胞蛋白质组图谱（可在www.fitsarpro.appspot.com获取）。结果表明使用FITSAR评估细胞类型特异性假说的可行性。我们的整体方法提供了一个可用于确定特定细胞类型中蛋白质存在情况的平台，并强调了使用蛋白质兼容技术来解析因细胞异质性而复杂的系统的必要性。
