Multiplexed protein profiling reveals spatial subcellular signaling networks.
Multiplexed protein profiling reveals spatial subcellular signaling networks.
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DOI：
10.1016/j.isci.2022.104980
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发表时间：
2022-09-16
期刊：
影响因子：
5.8
通讯作者：
Coskun, Ahmet F.
中科院分区：
文献类型：
--
作者：
Cai, Shuangyi;Hu, Thomas;Venkatesan, Mythreye;Allam, Mayar;Schneider, Frank;Ramalingam, Suresh S.;Sun, Shi-Yong;Coskun, Ahmet F.
蛋白质-蛋白质相互作用网络在许多疾病的多基因失调中发生改变。基于图像的蛋白质多路复用揭示了信号通路，以解剖细胞间的异质性，以前被批量检测所掩盖。在此，我们提出了一种快速多路免疫荧光（RapMIF）方法，在亚细胞分辨率下测量来自培养物和组织的多达25路空间蛋白质图谱，在33路多路像素级簇中提供272对和1360三蛋白信号状态。RapMIF流水线在单个平台上自动对生物标本进行染色、漂白和成像。RapMIF显示WNT/β-catenin信号在抑制AKT/mTOR通路时上调。亚细胞蛋白图像显示了单个细胞中的易位模式、空间受体不连续和亚细胞信号簇。信号网络在药物反应培养中表现出信号蛋白的空间再分布。机器学习分析从相互连接的信号蛋白图像预测磷酸化的β-catenin表达。RapMIF是精确治疗设计的理想信号发现方法。像素聚类图揭示信号蛋白的邻域奥西替尼下调p-AKT并抑制细胞增殖和存活多路分析解剖信号网络用于药物反应预测生物科学；生物技术;生物科学研究方法；生物学实验方法
Protein-protein interaction networks are altered in multi-gene dysregulations in many disorders. Image-based protein multiplexing sheds light on signaling pathways to dissect cell-to-cell heterogeneity, previously masked by the bulk assays. Herein, we present a rapid multiplexed immunofluorescence (RapMIF) method measuring up to 25-plex spatial protein maps from cultures and tissues at subcellular resolution, providing combinatorial 272 pairwise and 1,360 tri-protein signaling states across 33 multiplexed pixel-level clusters. The RapMIF pipeline automated staining, bleaching, and imaging of the biospecimens in a single platform. RapMIF showed that WNT/β-catenin signaling upregulated upon the inhibition of the AKT/mTOR pathway. Subcellular protein images demonstrated translocation patterns, spatial receptor discontinuity, and subcellular signaling clusters in single cells. Signaling networks exhibited spatial redistribution of signaling proteins in drug-responsive cultures. Machine learning analysis predicted the phosphorylated β-catenin expression from interconnected signaling protein images. RapMIF is an ideal signaling discovery approach for precision therapy design. Pixel clustering maps revealed the neighborhood of signaling proteins Osimertinib downregulated p-AKT and inhibited cell proliferation and survival Multiplexed profiling dissects signaling networks for drug response prediction Biological sciences; Biotechnology; Biological sciences research methodologies; Biology experimental methods
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影响因子：
4.8
作者：
He, B;You, L;Jablons, DM
通讯作者：
Jablons, DM
DOI：
10.18632/aging.103475
发表时间：
2020-09-14
期刊：
Aging
影响因子：
--
作者：
Bu J;Zhang P;Zhu K;Yan Y;Shi B;Wang J;Xu S
通讯作者：
Xu S
影响因子：
48
作者：
Giesen, Charlotte;Wang, Hao A. O.;Bodenmiller, Bernd
通讯作者：
Bodenmiller, Bernd
影响因子：
6.2
作者：
Gu, Jiajia;Yao, Weilong;Sun, Shi-Yong
通讯作者：
Sun, Shi-Yong
影响因子：
7.7
作者：
Civenni, G;Holbro, T;Hynes, NE
通讯作者：
Hynes, NE