Development and validation of a novel assay to identify radiosensitizers that target nucleophosmin 1. Issue 13 (1st July 2015)

• Record Type: Journal Article
• Authors: Penthala, Narsimha R.; Crooks, Peter A.; Freeman, Michael L.; Sekhar, Konjeti R.
• Is Part Of: Bioorganic & medicinal chemistry. Volume 23:Issue 13(2015)
• Page Start: 3681
• Page End: 3686
• Publication Date: 2015-07-01
• Subjects: Radiosensitizers -- Assay development -- Cancer -- DNA repair -- Nucleophosmin 1 -- NPM1 -- YTR107 -- PNR605 -- Indole analogs -- Radiation -- Lung cancer; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; Biochemistry -- Periodicals; Chemistry, Clinical -- Periodicals; Chemistry, Organic -- Periodicals; Chimie bio-organique -- Périodiques; Chimie pharmaceutique -- Périodiques; 615.19
• DOI: 10.1016/j.bmc.2015.04.018
• Languages: English
• ISSNs: 0968-0896

Abstract

Graphical abstract: Novel Western blot assay was developed and tested for its applicability in identifying radiosensitizers that target nucleophosmin 1 (NPM1). Extractability of pNPM1 was used as a measurement for identification of active compounds from a library of analogs synthesized based on the lead molecule, YTR107. Abstract: A series of indole analogs that are synthesized using the scaffold of a potent radiosensitizer, YTR107, were tested for their ability to alter the solubility of phosphorylated nucleophosmin 1 (pNPM1). NPM1 is critical for DNA double strand break (DSB) repair. In response to formation of DNA DSBs, phosphorylated T199 NPM1 binds to ubiquitinated chromatin, in a RNF8/RNF168-dependent manner, forming irradiation-induced foci (IRIF) that promote repair of DNA DSBs. A Western blot assay was developed using lead molecule, YTR107, for the purpose of screening newly synthesized molecules that target pNPM1 in irradiated cells. A colony formation assay was used to demonstrate the radiosensitization properties of the compounds. Compounds that enhanced the extractability of pNPM1 upon radiation treatment possessed radiosensitization properties.