Oncotarget

Research Papers:

VX-984 is a selective inhibitor of non-homologous end joining, with possible preferential activity in transformed cells

Atif J. Khan _, Sarah M. Misenko, Aditya Thandoni, Devora Schiff, Sachin R. Jhawar, Samuel F. Bunting and Bruce G. Haffty

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Oncotarget. 2018; 9:25833-25841. https://doi.org/10.18632/oncotarget.25383

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Abstract

Atif J. Khan1,2, Sarah M. Misenko3, Aditya Thandoni2, Devora Schiff2, Sachin R. Jhawar2, Samuel F. Bunting3 and Bruce G. Haffty2

1Department of Radiation Oncology, Memorial Sloan Kettering Cancer Center, New York, NY 10011, USA

2Department of Radiation Oncology, Rutgers-Robert Wood Johnson Medical School, Piscataway, NJ 08854, USA

3Department of Molecular Biology and Biochemistry, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA

Correspondence to:

Atif J. Khan, email: khana7@mskcc.org

Keywords: DNA-PK; non-homologous end joining; DNA repair; double-strand break repair; radiation therapy

Received: December 05, 2017     Accepted: April 25, 2018     Published: May 25, 2018

ABSTRACT

Purpose: DNA double-strand breaks (DSBs) can be repaired by non-homologous end joining (NHEJ) or homologous recombination (HR). We demonstrate the selectivity of VX-984, a DNA-PK inhibitor, using assays not previously reported.

Experimental Design: The class switch recombination assay (CSR) in primary B cells was used to measure efficiency of NHEJ. A cellular reporter assay (U2OS EJ-DR) was used to assess the efficiency of HR and NHEJ in cells treated with VX-984. Immunofluorescence assays (IF) evaluated γ-H2AX foci for DSB repair kinetics in human astrocytes and T98G glioma cells. Western blotting was used to evaluate phosphorylation of DNA-PKcs substrates.

Results: We found a dose-dependent reduction in CSR efficiency with VX-984, and through the EJ-DR assay, dramatic dose-dependent increases in HR and mNHEJ. Immunofluorescence assays showed an inability of malignant cells to resolve γ-H2AX foci in the presence of VX-984. Radiation-induced phosphorylation of DNA-PK substrates was further reduced by treatment with VX-984.

Conclusions: VX-984 efficiently inhibits NHEJ, resulting in compensatory increases in alternative repair pathways, increases DSBs, and appears to affect transformed cells preferentially.


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