A High-Throughput Assay for Small Molecule Destabilizers of the KRAS Oncoprotein

作者：Carver Joseph; Dexheimer Thomas S; Hsu Dennis; Weng Meng Tzu; Smith Jordan L; Guha Rajarshi; Jadhav Ajit; Simeonov Anton^*; Luo Ji

来源：PLos One, 2014, 9(8): e103836.

DOI：10.1371/journal.pone.0103836

Mutations in the Ras family of small GTPases, particularly KRAS, occur at high frequencies in cancer and represent a major unmet therapeutic need due to the lack of effective targeted therapies. Past efforts directed at inhibiting the activity of the Ras oncoprotein have proved difficult. We propose an alternative approach to target Ras by eliminating Ras protein from cells with pharmacological means. In this study, we developed a cell-based, high-content screening platform to identify small molecules that could promote the degradation of the KRAS oncoprotein. We generated an EGFP-KRAS(G12V) fluorescence reporter system and implemented it for automated screening in 1536-well plates using high-throughput cellular imaging. We screened a library of clinically relevant compounds at wide dose range and identified Ponatinib and AMG-47a as two candidate compounds that selectively reduced the levels of EGFP-KRAS(G12V) protein but did not affect EGFP protein in cells. This proof-of-principle study demonstrates that it is feasible to use a high-throughput screen to identify compounds that promote the degradation of the Ras oncoprotein as a new approach to target Ras.

• 出版日期2014-8-5