Molecular imaging of epidermal growth factor receptor kinase activity
Molecular imaging of epidermal growth factor receptor kinase activity
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DOI：
10.1016/j.ab.2011.05.040
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发表时间：
2011-10-01
影响因子：
2.9
通讯作者：
Rehemtulla, Alnawaz
中科院分区：
文献类型：
--
作者：
Khan, Amjad P.;Contessa, Joseph N.;Rehemtulla, Alnawaz
表皮生长因子受体（埃格）是一种受体酪氨酸激酶，通常在不同的肿瘤类型中发生改变，导致异常调节的激酶活性和下游信号级联的过度激活，包括细胞增殖、分化和迁移。为了在真实的时间和活细胞和动物中研究EGFR信号传导事件，在这里，我们描述了一种多结构域嵌合报告基因，其生物发光可用作EGFR激酶活性的替代物。在鳞状细胞癌细胞（UMSCC1）中开发了这种基于端粒酶的报告基因，以生成用于EGFR成像的癌症治疗模型。报告基因被设计为EGFR的磷酸化底物，并在未磷酸化时重建荧光素酶活性，从而提供EGFR抑制的稳健指示。我们在体外验证了报告基因，并证明其活性可通过厄洛替尼抑制EGFR酪氨酸激酶或表皮生长因子激活受体进行差异调节。进一步的体内实验证实了对异种移植物中EGFR酪氨酸激酶活性的定量和动态监测。从这些研究中获得的结果提供了独特的见解药物的药代动力学和药效学的药物调节埃格活性，揭示了该报告在评估药物的可用性和细胞靶向活细胞和小鼠模型的有用性。(C)2011 Elsevier Inc. All rights reserved.
Epidermal growth factor receptor (EGER), a receptor tyrosine kinase, is commonly altered in different tumor types, leading to abnormally regulated kinase activity and excessive activation of downstream signaling cascades, including cell proliferation, differentiation, and migration. To investigate the EGFR signaling events in real time and in living cells and animals, here we describe a multidomain chimeric reporter whose bioluminescence can be used as a surrogate for EGFR kinase activity. This luciferase-based reporter was developed in squamous cell carcinoma cells (UMSCC1) to generate a cancer therapy model for imaging EGFR. The reporter is designed to act as a phosphorylated substrate of EGFR and reconstitutes luciferase activity when it is not phosphorylated, thereby providing a robust indication of EGFR inhibition. We validated the reporter in vitro and demonstrated that its activity could be differentially modulated by EGFR tyrosine kinase inhibition with erlotonib or receptor activation with epidermal growth factor. Further experiments in vivo demonstrated quantitative and dynamic monitoring of EGFR tyrosine kinase activity in xenograft. Results obtained from these studies provide unique insight into pharmacokinetics and pharmacodynamics of agents that modulate EGER activity, revealing the usefulness of this reporter in evaluating drug availability and cell targeting in both living cells and mouse models. (C) 2011 Elsevier Inc. All rights reserved.