A Live-Cell Screen for Altered Erk Dynamics Reveals Principles of Proliferative Control.
A Live-Cell Screen for Altered Erk Dynamics Reveals Principles of Proliferative Control.
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DOI：
10.1016/j.cels.2020.02.005
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发表时间：
2020-03-25
期刊：
影响因子：
9.3
通讯作者：
Toettcher JE
中科院分区：
文献类型：
--
作者：
Goglia AG;Wilson MZ;Jena SG;Silbert J;Basta LP;Devenport D;Toettcher JE
在细胞信号传导中广泛观察到复杂的、随时间变化的反应，但具体的动态是如何产生或调节的却很大程度上未知。一个主要障碍是高通量筛选通常与用于监测动态的活细胞测定不兼容。在这里，我们通过筛选 429 种激酶抑制剂库并在 5 小时内监测超过 80,000 个原代小鼠角质形成细胞中的 Erk 活性来应对这一挑战。我们的筛选揭示了已知和未表征的 Erk 动力学调节剂，包括增加 Erk 脉冲频率和整体活性的非 EGFR 受体酪氨酸激酶 (RTK) 抑制剂。通过药物治疗和直接光遗传学控制，我们证明药物诱导的 Erk 动力学变化会改变细胞增殖的条件。我们的工作打开了使用活细胞生物传感器进行高通量筛选的大门，并揭示了细胞增殖整合了来自 Erk 动力学的信息以及其他许可线索。戈利亚等人。通过筛选 429 种激酶抑制剂库并在 5 小时内监测超过 80,000 个原代小鼠角质形成细胞中的 Erk 活性，确定了 ERK 动态调节剂。他们鉴定了已知和未表征的调节剂，包括增加 Erk 脉冲频率和整体活性的非 EGFR 受体酪氨酸激酶 (RTK) 抑制剂。他们的工作打开了使用活细胞生物传感器进行高通量筛选的大门，并揭示了细胞增殖整合了来自 Erk 动力学的信息以及其他许可线索。
Complex, time-varying responses have been observed widely in cell signaling, but how specific dynamics are generated or regulated is largely unknown. One major obstacle has been that high-throughput screens are typically incompatible with the live-cell assays used to monitor dynamics. Here, we address this challenge by screening a library of 429 kinase inhibitors and monitoring Erk activity over 5 hours in more than 80,000 single primary mouse keratinocytes. Our screen revealed both known and uncharacterized modulators of Erk dynamics, including inhibitors of non-EGFR receptor tyrosine kinases (RTKs) that increased Erk pulse frequency and overall activity. Using drug treatment and direct optogenetic control, we demonstrate that drug-induced changes to Erk dynamics alter the conditions under which cells proliferate. Our work opens the door to high-throughput screens using live-cell biosensors and reveals that cell proliferation integrates information from Erk dynamics as well as additional permissive cues. Goglia et al. identified modulators of ERK dynamics by screening a library of 429 kinase inhibitors and monitoring Erk activity over 5 hours in more than 80,000 single primary mouse keratinocytes. They identified both known and uncharacterized modulators, including inhibitors of non-EGFR receptor tyrosine kinases (RTKs) that increased Erk pulse frequency and overall activity. Their work opens the door to high-throughput screens using live-cell biosensors and reveals that cell proliferation integrates information from Erk dynamics as well as additional permissive cues.
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影响因子：
64.5
作者：
Heidorn SJ;Milagre C;Whittaker S;Nourry A;Niculescu-Duvas I;Dhomen N;Hussain J;Reis-Filho JS;Springer CJ;Pritchard C;Marais R
通讯作者：
Marais R
影响因子：
46.9
作者：
通讯作者：
--
影响因子：
64.5
作者：
Feldman, David;Singh, Avtar;Blainey, Paul C.
通讯作者：
Blainey, Paul C.
影响因子：
9.3
作者：
Gillies TE;Pargett M;Minguet M;Davies AE;Albeck JG
通讯作者：
Albeck JG
DOI：
10.1073/pnas.1417910112
发表时间：
2015-01-06
影响因子：
11.1
作者：
Guntas, Gurkan;Hallett, Ryan A.;Kuhlman, Brian
通讯作者：
Kuhlman, Brian