Deciphering the History of ERK Activity from Fixed-Cell Immunofluorescence Measurements.
Deciphering the History of ERK Activity from Fixed-Cell Immunofluorescence Measurements.
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从固定细胞免疫荧光测量中解读 ERK 活性的历史。
DOI：
10.1101/2024.02.16.580760
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Albeck,John
中科院分区：
文献类型：
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作者：
Ram,Abhineet;Pargett,Michael;Choi,Yongin;Murphy,Devan;Cabel,Markhus;Kosaisawe,Nont;Quon,Gerald;Albeck,John
RAS/ERK 通路在许多癌症的诊断和治疗中发挥着核心作用。 ERK 活性在单个细胞内高度动态，并通过效应蛋白（包括 c-Myc、c-Fos、Fra-1 和 Egr-1）驱动细胞增殖、代谢和其他过程。这些蛋白质对 ERK 活性的动态敏感，但尚不清楚单个细胞中的 ERK 活性模式在多大程度上决定效应蛋白表达，或者效应器表达模式中嵌入了多少有关 ERK 动态的信息。在这里，我们使用活细胞生物传感器测量 ERK 活性来评估这些关系，并与该通路下游靶蛋白的免疫荧光染色相结合。将这些数据集与线性回归、机器学习和微分方程模型相结合，我们开发了免疫荧光数据的解释框架，其中 Fra-1 和 pRb 水平意味着 ERK 信号传导的长期激活，而 Egr-1 和 c-Myc 则表明最近的激活。对多种癌细胞系的分析揭示了恶性细胞中 ERK 活性与细胞状态之间的扭曲关系。我们表明，该框架可以从异质群体内的效应蛋白染色推断各种类型的 ERK 动态，为注释固定细胞内的 ERK 动态提供基础。
The RAS/ERK pathway plays a central role in diagnosis and therapy for many cancers. ERK activity is highly dynamic within individual cells and drives cell proliferation, metabolism, and other processes through effector proteins including c-Myc, c-Fos, Fra-1, and Egr-1. These proteins are sensitive to the dynamics of ERK activity, but it is not clear to what extent the pattern of ERK activity in an individual cell determines effector protein expression, or how much information about ERK dynamics is embedded in the pattern of effector expression. Here, we evaluate these relationships using live-cell biosensor measurements of ERK activity, multiplexed with immunofluorescence staining for downstream target proteins of the pathway. Combining these datasets with linear regression, machine learning, and differential equation models, we develop an interpretive framework for immunofluorescence data, wherein Fra-1 and pRb levels imply long-term activation of ERK signaling, while Egr-1 and c-Myc indicate more recent activation. Analysis of multiple cancer cell lines reveals a distorted relationship between ERK activity and cell state in malignant cells. We show that this framework can infer various classes of ERK dynamics from effector protein stains within a heterogeneous population, providing a basis for annotating ERK dynamics within fixed cells.