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Development of Cell-based High Throughput Luminescence Assay for Drug Discovery in Inhibiting OCT4 and MAPKAPK2 Interaction
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  • Sung-Jen Wei,
  • In-Hyoung Yang,
  • Gloria Martinez,
  • Shengping Yang,
  • Eun Jeong Cho,
  • Kevin Dalby,
  • Min Kang
Sung-Jen Wei
Texas Tech University Health Sciences Center
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In-Hyoung Yang
Texas Tech University Health Sciences Center
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Gloria Martinez
Texas Tech University Health Sciences Center
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Shengping Yang
Pennington Biomedical Research Center
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Eun Jeong Cho
The University of Texas at Austin College of Pharmacy
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Kevin Dalby
The University of Texas at Austin College of Pharmacy
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Min Kang
Texas Tech University Health Sciences Center
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Abstract

Overexpression of c-MYC protein without genomic amplification occurs in multiple cancers, including neuroblastoma and small cell lung cancer. In searching for the mechanisms of c-MYC protein overexpression, we demonstrated that the transcription factor, OCT4 mediates c-MYC transcriptional activation in progressive disease neuroblastoma. Subsequently, we identified two kinases, MAPKAPK2 (MK2) and DNA-PK, which are predicted to bind and phosphorylate OCT4 at S111 and S93 residues, respectively. Based on these novel observations, we developed a cell-based luminescence assay to screen and identify compounds that inhibit the interactions between MK2 and OCT4. By screening 79,671 compounds, we identified 65 compounds we designated as “hits”. Using a two-step validation of co-immunoprecipitation and pOCT4S111 detection, the compounds were further narrowed down to three for further studies. The three compounds were tested for their ability to inhibit kinase activity, in vitro cytotoxic activity, and anti-inflammatory activity. In conclusion, we developed a cell-based luminescence assay for the discovery of new agents targeting the c-MYC transcriptional activation pathway. Screening and subsequent validation identified a small number of compounds for further development.

Peer review status:POSTED

20 Oct 2020Submitted to Biotechnology and Bioengineering
21 Oct 2020Assigned to Editor
21 Oct 2020Submission Checks Completed