Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel

Haiyan Sun; Ying Wei; Huayun Deng; Qiaojie Xiong; Min Li; Joydeep Lahiri; Ye Fang

doi:10.1038/srep04934

Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel

Haiyan Sun, Ying Wei, Huayun Deng, Qiaojie Xiong, Min Li, Joydeep Lahiri, Ye Fang

Research output: Contribution to journal › Article › peer-review

21 Scopus citations

Abstract

Current technologies for studying ion channels are fundamentally limited because of their inability to functionally link ion channel activity to cellular pathways. Herein, we report the use of label-free cell phenotypic profiling to decode the composition and signaling of an endogenous ATP-sensitive potassium ion channel (K_ATP) in HepG2C3A, a hepatocellular carcinoma cell line. Label-free cell phenotypic agonist profiling showed that pinacidil triggered characteristically similar dynamic mass redistribution (DMR) signals in A431, A549, HT29 and HepG2C3A, but not in HepG2 cells. Reverse transcriptase PCR, RNAi knockdown, and K_ATP blocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 K_ATP channels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated K_ATP channels trigger signaling through Rho kinase and Janus kinase-3, and cause actin remodeling. The results are the first demonstration of a label-free methodology to characterize the composition and signaling of an endogenous ATP-sensitive potassium ion channel.

Original language	English (US)
Article number	4934
Journal	Scientific reports
Volume	4
DOIs	https://doi.org/10.1038/srep04934
State	Published - May 12 2014
Externally published	Yes

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title = "Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel",

abstract = "Current technologies for studying ion channels are fundamentally limited because of their inability to functionally link ion channel activity to cellular pathways. Herein, we report the use of label-free cell phenotypic profiling to decode the composition and signaling of an endogenous ATP-sensitive potassium ion channel (KATP) in HepG2C3A, a hepatocellular carcinoma cell line. Label-free cell phenotypic agonist profiling showed that pinacidil triggered characteristically similar dynamic mass redistribution (DMR) signals in A431, A549, HT29 and HepG2C3A, but not in HepG2 cells. Reverse transcriptase PCR, RNAi knockdown, and KATP blocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 KATP channels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated KATP channels trigger signaling through Rho kinase and Janus kinase-3, and cause actin remodeling. The results are the first demonstration of a label-free methodology to characterize the composition and signaling of an endogenous ATP-sensitive potassium ion channel.",

author = "Haiyan Sun and Ying Wei and Huayun Deng and Qiaojie Xiong and Min Li and Joydeep Lahiri and Ye Fang",

note = "Funding Information: Supplementary information accompanies this paper at http://www.nature.com/ scientificreports Competing financial interests: Y.F., Y.W., H.D. and J.L. are employee of Corning Incorporated. DMR assay is patented. EpicH system is commercial product from Corning Incorporated. This work was partially supported by the National Institutes of Health Grant 5U54MH084691. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.",

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doi = "10.1038/srep04934",

language = "English (US)",

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T1 - Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel

AU - Sun, Haiyan

AU - Wei, Ying

AU - Deng, Huayun

AU - Xiong, Qiaojie

AU - Li, Min

AU - Lahiri, Joydeep

AU - Fang, Ye

N1 - Funding Information: Supplementary information accompanies this paper at http://www.nature.com/ scientificreports Competing financial interests: Y.F., Y.W., H.D. and J.L. are employee of Corning Incorporated. DMR assay is patented. EpicH system is commercial product from Corning Incorporated. This work was partially supported by the National Institutes of Health Grant 5U54MH084691. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

PY - 2014/5/12

Y1 - 2014/5/12

N2 - Current technologies for studying ion channels are fundamentally limited because of their inability to functionally link ion channel activity to cellular pathways. Herein, we report the use of label-free cell phenotypic profiling to decode the composition and signaling of an endogenous ATP-sensitive potassium ion channel (KATP) in HepG2C3A, a hepatocellular carcinoma cell line. Label-free cell phenotypic agonist profiling showed that pinacidil triggered characteristically similar dynamic mass redistribution (DMR) signals in A431, A549, HT29 and HepG2C3A, but not in HepG2 cells. Reverse transcriptase PCR, RNAi knockdown, and KATP blocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 KATP channels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated KATP channels trigger signaling through Rho kinase and Janus kinase-3, and cause actin remodeling. The results are the first demonstration of a label-free methodology to characterize the composition and signaling of an endogenous ATP-sensitive potassium ion channel.

AB - Current technologies for studying ion channels are fundamentally limited because of their inability to functionally link ion channel activity to cellular pathways. Herein, we report the use of label-free cell phenotypic profiling to decode the composition and signaling of an endogenous ATP-sensitive potassium ion channel (KATP) in HepG2C3A, a hepatocellular carcinoma cell line. Label-free cell phenotypic agonist profiling showed that pinacidil triggered characteristically similar dynamic mass redistribution (DMR) signals in A431, A549, HT29 and HepG2C3A, but not in HepG2 cells. Reverse transcriptase PCR, RNAi knockdown, and KATP blocker profiling showed that the pinacidil DMR is due to the activation of SUR2/Kir6.2 KATP channels in HepG2C3A cells. Kinase inhibition and RNAi knockdown showed that the pinacidil activated KATP channels trigger signaling through Rho kinase and Janus kinase-3, and cause actin remodeling. The results are the first demonstration of a label-free methodology to characterize the composition and signaling of an endogenous ATP-sensitive potassium ion channel.

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Label-free cell phenotypic profiling decodes the composition and signaling of an endogenous ATP-sensitive potassium channel

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