figure posted on 2023-10-16, 14:20 authored by Helena Folly-Kossi, Joshua D. Graves, Lidija A. Wilhelms Garan, Fang-Tsyr Lin, Weei-Chin Lin
DNA2 inhibitor d16 decreases HR repair. A, Quantification of HR activity in HEK293T cells using the TLR assay after treatment with d16 (20 µmol/L). **, P < 0.01 (two-tailed t test). A representative set of flow profiles are shown in Supplementary Fig. S6. B, MDAH-2774 cells were treated with d16, RU or both for 8 hours. After fixation and permeabilization, immunostaining was performed with an anti-RAD51 rabbit antibody, followed by Texas Red X-conjugated anti-rabbit secondary antibody. Nuclei were stained with Hoechst 33258. Pictures shown are representative images at 100X magnification. C, Quantification of RAD51 foci-positive cells. ***, P < 0.001 (two-tailed t test). D, Immunoblotting shows the expression of DNA2, Rad51, γH2AX, or β-actin in the whole-cell lysates. The Ub-γH2AX signals were quantified using NIH ImageJ software.
HHS | National Institutes of Health (NIH)
U.S. Department of Defense (DOD)
Rivkin Center for Ovarian Cancer (Rivkin Center)
ARTICLE ABSTRACTThe tumor suppressor p53 promotes tumor-suppressive activities including cell-cycle inhibition, apoptosis, senescence, autophagy, and DNA repair. However, somatic mutations in the TP53 gene are one of the most common alterations in human cancers. We previously showed that mutant p53 (mutp53) can bind TopBP1, an ATR activator, to attenuate its ATR-activating function. A partially defective ATR function caused by mutp53 makes cancer cells more vulnerable to inhibitors of other TopBP1-independent ATR activators, such as DNA2. DNA2 plays a role in homologous recombination (HR) repair by resecting DNA ends in double-strand breaks and preparing them for invasion of homologous duplex. Here we identify a new DNA2 inhibitor, namely d16, and show that d16 exhibits anticancer activities and overcomes chemotherapy resistance in mutp53-bearing cancers. Similar to DNA2 depletion, d16 treatment results in cell-cycle arrest mainly at S-phase. Moreover, reexpression of mutp53 in a p53-null cancer cell line makes cells more vulnerable to d16-mediated inhibition of ATR activity. As d16 also inhibits HR, a combination of d16 and PARP inhibitors displays synergistic induction of cell death. DNA2 is often overexpressed in cancer, particularly in cancer cells harboring mutp53. Overexpression of DNA2 is associated with poor outcome in ovarian cancer. Overall, our results provide a rationale to target DNA2 as a new synthetic lethality approach in mutp53-bearing cancers, and further extend the benefit of PARP inhibitors beyond BRCA-mutated cancers.
This study identifies a new DNA2 inhibitor as a synthetic lethal targeted therapy for mutp53-harboring cancers, and provides a new therapeutic strategy by combining DNA2 inhibitors with PARP inhibitors for these cancers.