Development and validation of a novel assay to identify radiosensitizers that target nucleophosmin 1.
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.
Department of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Narsimha R
NR
Penthala
Department of Pharmaceutical Sciences, College of Pharmacy, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Peter A
PA
Crooks
Department of Radiation Oncology, Vanderbilt University School Medicine, Nashville, TN 37232, USA.
Michael L
ML
Freeman
Department of Radiation Oncology, Vanderbilt University School Medicine, Nashville, TN 37232, USA. Electronic address: raja.konjeti@vanderbilt.edu.
Konjeti R
KR
Sekhar
D001463:Barbiturates / Q000494:pharmacology*; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D018385:Centrosome / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects; D053903:DNA Breaks, Double-Stranded / Q000528:radiation effects*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects; D005720:Gamma Rays; D006801:Humans; D007211:Indoles / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000090243:Nucleophosmin; D010766:Phosphorylation; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology*; D014410:Tumor Stem Cell Assay; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism
C569647:5-((N-benzyl-1H-indol-3-yl)methylene)pyrimidine-2,4,6(1H,3H,5H)trione; D001463:Barbiturates; D002843:Chromatin; D007211:Indoles; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D011838:Radiation-Sensitizing Agents; D025801:Ubiquitin; D000090243:Nucleophosmin
Assay development; Cancer; DNA repair; Indole analogs; Lung cancer; NPM1; Nucleophosmin 1; PNR605; Radiation; Radiosensitizers; YTR107