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. 2022 Aug 5;28(21):4724–4736. doi: 10.1158/1078-0432.CCR-22-0301

Figure 2.

Figure 2. AZD5305 selectively targets cancer cells with HRR-Deficiency, inducing DNA damage accumulation and cell cycle arrest. A, Colony formation assays of DLD-1 isogenic cell line pair (WT in dotted line and BRCA2−/− in solid line) treated with AZD5305 in dose response (A, left) Curves are the mean of six independent experiments, error bars indicate ±SEM. (A, bottom) Representative images of clonogenic assays. (A, right) Bar chart representing the differential activity of each PARPi on the BRCA2−/- isogenic cell line pair. Bars are the IC50 ratio of WT over the BRCA2−/− cells for each indicated PARPi. (B, left). Colony formation assays performed in SKOV-3 isogenic pairs: WT (dotted line, empty circle), BRCA2-KO, PALB2-KO and RAD51C-KO (solid lines and full symbols, as indicated) treated with a dose response of AZD5305. Curves are the mean of three independent experiments, error bars indicate ±SEM. (B, right) Bar chart representing the differential activity of each compound on each isogenic cell line pair. Bars are the IC50 ratio of WT over the indicated KO isogenic pair, measured for each indicated PARPi. (C, left) γH2AX measured by immunofluorescence after 72-hour treatments with a dose response of AZD5305 in DLD-1 BRCA2−/− (solid line and full symbol) or WT cells (dotted line and empty symbol). Curves are the mean of three independent experiments, error bars indicate ±SEM. (C, right), representative immunofluorescence images of DLD-1 BRCA2−/− cells treated with 10 nmol/L of AZD5305 (bottom) or untreated (top). D, Cell cycle analysis by flow cytometry of DLD-1 BRCA2−/− (left) and WT cells (right) treated with AZD5305 for 48 hours at indicated concentrations. Bar charts represent the distribution of cells in the different cell cycle phases, G1, S, and G2; below are representative cell cycle scatter plots, with DNA and EdU intensity signal on x- and y- axes, respectively.

AZD5305 selectively targets cancer cells with HRR-deficiency, inducing DNA damage accumulation and cell-cycle arrest. A, Left, Colony formation assays of DLD-1 isogenic cell line pair (WT in dotted line and BRCA2−/− in solid line) treated with AZD5305 in dose–response. Curves are the mean of six independent experiments; error bars indicate ±SEM. Bottom, Representative images of clonogenic assays. Right, Bar chart representing the differential activity of each PARPi on the BRCA2−/− isogenic cell line pair. Bars are the IC50 ratio of WT over the BRCA2−/− cells for each indicated PARPi. B, Left, Colony formation assays performed in SKOV-3 isogenic pairs: WT (dotted line, empty circle), BRCA2-KO, PALB2-KO, and RAD51C-KO (solid lines and full symbols, as indicated) treated with a dose–response of AZD5305. Curves are the mean of three independent experiments; error bars indicate ±SEM. Right, Bar chart representing the differential activity of each compound on each isogenic cell line pair. Bars are the IC50 ratio of WT over the indicated KO isogenic pair, measured for each indicated PARPi. C, Left, γH2AX measured by immunofluorescence after 72-hour treatments with a dose–response of AZD5305 in DLD-1 BRCA2−/− (solid line and full symbol) or WT cells (dotted line and empty symbol). Curves are the mean of three independent experiments; error bars indicate ±SEM. Right, representative immunofluorescence images of DLD-1 BRCA2−/− cells treated with 10 nmol/L of AZD5305 (bottom) or untreated (top). D, Cell-cycle analysis by flow cytometry of DLD-1 BRCA2−/− (left) and WT cells (right) treated with AZD5305 for 48 hours at indicated concentrations. Bar charts represent the distribution of cells in the different cell-cycle phases, G1, S, and G2; below are representative cell-cycle scatter plots, with DNA and EdU intensity signal on x- and y-axes, respectively.