Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: Telmisartan enhances olaparib efficacy in vitro and in vivo . ( a ) MTT viability of indicated cell lines treated with indicated concentrations of olaparib ± 10 µM telmisartan for 96 hours. P by two-way ANOVA. ( b ) Bliss synergy analysis of ID8agg cells conducted by SynergyFinder. Bliss scores >15 define synergy. ( c ) Immunoblots of indicated cell lines treated with 5 µM olaparib ± 10 µM telmisartan for 48 hours. ( d ) Flow cytometry analysis of DNA damage by γH2AX staining of splenocytes incubated in vitro with 10 µM telmisartan overnight. P by unpaired t-test. ANOVA, analysis of variance; MTT, 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyl-2H-tetrazolium bromide.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: In Vitro, In Vivo, Western Blot, Flow Cytometry, Staining, Incubation

Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: Telmisartan augments olaparib-induced G2/M cell cycle arrest independently of its ARB mechanism. ( a–b ) Representative histograms of flow cytometry propidium iodide cell cycle data from ( a ) ID8agg and ( b ) MC38 cells. ( c–d ) Propidium iodide cell cycle analysis of ( c ) ID8agg and ( d ) MC38 cells treated with 5 µM olaparib ± 10 µM telmisartan. P by unpaired t-test. ( e–f ) MTT viability of ( e ) 4T1 and ( f ) ID8agg cells treated with indicated concentrations of olaparib ± 10 µM of ARB drugs as indicated for 96 hours. P by two-way ANOVA. ANOVA, analysis of variance; ARB, angiotensin type II receptor blocker; MTT, 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyl-2H-tetrazolium bromide.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: Flow Cytometry, Cell Cycle Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: Telmisartan synergizes with olaparib independently of tumor PD-L1 and HR. ( a ) MTT viability of indicated genetic PD-L1-deficient cell lines incubated with 10 µM telmisartan and indicated concentrations of olaparib for 96 hours. P by two-way ANOVA. ( b ) Immunoblots of indicated genetic PD-L1-deficient cell lines incubated with 5 µM olaparib ± 10 µM telmisartan for 48 hours. ( c ) Immunoblots of cell lines treated with 5 µM olaparib ± 10 µM telmisartan for 48 hours. ( d ) qRT-PCR for CD274 of indicated cell lines incubated with 5 µM olaparib ± 10 µM telmisartan. P by unpaired t-test. ( e ) Immunoblots of indicated proteins in T24 cells treated with 5 µM olaparib ± 10 µM telmisartan for 48 hours. ( f ) Immunoblots of T24 cells incubated with 10 µM telmisartan ± 5 µM olaparib for 48 hours for indicated proteins. ( g ) U2OS cells engineered with indicated reporter plasmids to assess functional tests of HR, NHEJ through two distinct mechanisms, and single strand annealing by GFP expression detected throughflow cytometry. ANOVA, analysis of variance; green fluorescence protein (GFP); HR, homologous recombination; MTT, 3-(4,5-dimethylthiazol-2-yl)−2,5-diphenyl-2H-tetrazolium bromide; NHEJ, non-homologous end joining; PD-L1, programmed death ligand-1; qRT-PCR, quantitative real-time PCR.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: Incubation, Western Blot, Quantitative RT-PCR, Functional Assay, Expressing, Cytometry, Fluorescence, Homologous Recombination, Non-Homologous End Joining, Real-time Polymerase Chain Reaction

Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: Telmisartan increases cytosolic DNA to elicit STING signals and type I interferon production. ( a ) Immunoblots for indicated proteins in ID8agg and MC38 cells incubated with 5 µM olaparib ± 10 µM telmisartan for 48 hours. ( b ) Compiled qRT-PCR of cytosolic nuclear DNA isolated from ID8agg cells treated with 5 µM olaparib ± 10 µM telmisartan for 24 hours. Cytosolic genes assessed were GAPDH, PPIA, β-actin, and 18S, represented as individual data points. Values are the average of five biological replicates and are normalized to vehicle. P by one-way ANOVA with post hoc Tukey’s test. ( c ) qRT-PCR of indicated genes in MC38 cells incubated with 5 µM olaparib ± 10 µM telmisartan. P by unpaired t-test. ( d ) Immunoblots of MC38 cells incubated with 5 µM olaparib ± 10 µM telmisartan with 10 µg/mL isotype control or anti-IFNAR1 antibody for 48 hours. ANOVA, analysis of variance; qRT-PCR, quantitative real-time PCR.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: Western Blot, Incubation, Quantitative RT-PCR, Isolation, Control, Real-time Polymerase Chain Reaction

Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: Telmisartan improves olaparib efficacy in vivo through type I interferon signals. ( a ) Growth curves of subcutaneous MC38 tumors challenged in WT mice. Mice were treated with 50 mg/kg olaparib ± 5 mg/kg telmisartan daily starting day 3 post-challenge. P by two-way ANOVA. Data are shown as mean ± SEM. ( b ) Mice in ( a ) were sacrificed on day 23 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ( c ) Growth curves of subcutaneous MC38 tumors challenged into RAG1 KO mice treated with 50 mg/kg olaparib ± 5 mg/kg telmisartan starting on day 3 post-challenge. P by two-way ANOVA. Data are shown as mean ± SEM. ( d ) Mice in ( c ) were sacrificed on day 21 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ( e ) Growth curves of subcutaneous MC38 tumors challenged into WT C57/BL6 mice treated with 50 mg/kg olaparib and 5 mg/kg telmisartan ± αIFNAR1 or isotype control starting day 3 post-challenge. P by two-way ANOVA. Data are shown as mean ± SEM. ( f ) Mice in ( e ) were sacrificed on day 18 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ( g ) Growth curves of subcutaneous MC38 tumors challenged into WT mice or IFNAR1 KO mice treated with 50 mg/kg olaparib ± 5 mg/kg telmisartan starting day 3 post-challenge. P by two-way ANOVA. Data are shown as mean ± SEM. ( h ) Mice in ( g ) were sacrificed on day 17 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ( i ) Growth curves of subcutaneous MC38 tumors challenged into WT mice treated with αIFNAR1 ± 50 mg/kg olaparib alone or combined with 5 mg/kg telmisartan starting day 3 post-challenge. P by two-way ANOVA. Data are shown as mean±SEM. ( j ) Mice in ( i ) were sacrificed on day 17 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ( k ) 0.5×10 6 CTRL (STING + ) and STING KO MC38 tumor cells were challenged subcutaneously into WT mice. Mice were treated with 50 mg/kg olaparib ± 5 mg/kg telmisartan starting on day 5 post-challenge. P by two-way ANOVA. Data are shown as mean ± SEM. ( l ) Mice in ( k ) were sacrificed on day 17 post-challenge. Tumors were excised and weighed. P by unpaired t-test. ANOVA, analysis of variance; WT, wild-type.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: In Vivo, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: Telmisartan increases olaparib efficacy in homologous recombination proficient tumors by augmenting type I interferon production

doi: 10.1136/jitc-2025-012426

Figure Lengend Snippet: αIFNAR1 reduces antitumor immunity induced by telmisartan with olaparib in vivo . ( a–l ) Immune profiling by flow cytometry of tumors excised from mice in . Populations assessed include ( a ) IFNγ + CD8 + T cells, ( b ) IFN-γ MFI in CD8 + T cells, ( c ) granzyme B (GzB) + CD8+ T cells, ( d ) GzB MFI in CD8 + T cells, ( e ) IFN-γ + γδT cells, ( f ) IFN-γ MFI in γδT cells, ( g ) GzB + γδT cells, ( h ) GzB MFI in γδT cells, ( i ) IFN-γ + NK cells, ( j ) IFN-γ MFI in NK cells, ( k ) GzB + NK cells, and ( l ) GzB MFI in NK cells. P by unpaired t-test. IFN, interferon; GzB, granzyme B; MFI, mean fluorescence intensity; NK, natural killer; TCR, T-cell receptor.

Article Snippet: Olaparib and telmisartan were purchased from MedChemExpress.

Techniques: In Vivo, Flow Cytometry, Fluorescence

Journal: bioRxiv

Article Title: ATM safeguards DNA replication at endogenous base lesions

doi: 10.64898/2026.03.20.713105

Figure Lengend Snippet: a,b, Survival of the indicated BARD1 AID/AID HCT-116 cell lines exposed to the indicated doses of olaparib ( a ) or cisplatin ( b ). Resazurin cell viability assay, n=3 biological experiments, mean ± s.d. c, Immunofluorescence imaging-based quantification of RAD51 foci in indicated BARD1 AID/AID HCT-116 cell lines following 24 h treatments with olaparib ( n=2 biological experiments, >200 cells per condition). Horizontal bars, boxes and whiskers denote median, 25–75, and 5–95 percentile values, respectively. Significance, two-sided Kruskal–Wallis H test with Dunn’s correction for multiple comparisons. ****P ≤ 0.0001; ns non-significant. d,e, Immunofluorescence imaging of PAR levels in indicated HCT-116 cells treated with 1 mM PARGi or mock (DMSO) for 45 min and pulse-labelled with EdU (20 min, 10 µM). Representative cells ( d ), EdU -positive and -negative nuclei are outlined in yellow and white, respectively. Superplots ( e ), integrated PAR intensity per nucleus, n=3 biological experiments (each color denotes one replicate, bars indicate median and colored squares indicate median PAR signal per replicate). Significance, two-way ANOVA mixed model with repeat measures and Sidak’s correction for multiple comparisons. ****P ≤ 0.0001, ***P ≤ 0.001 **P ≤ 0.01. f,g DNA damage quantification in the indicated BARD1 AID/AID HCT-116 cells by comet assay under alkaline or neutral conditions. Representative images ( f ), and Scatter plots ( g ), show comet tail moments across n=3 biological experiments, each comprising 80-100 cells and represented in a different color. Horizontal bars, median values. Significance, two sided Kruskal-Wallis H test with Dunn’s correction for multiple comparisons. ****P ≤ 0.0001. h, Experimental design for i,j using splenic B cells. i,j Alkaline comet tail moments in quiescent resting and stimulated cycling cultures of splenic B cells from indicated mice. Scatter plot ( i ), each color represents one of n = 3 biological experiments; horizontal bars denote medians. Significance, Mann-Whitney t-test, ****P ≤ 0.0001; ns, non-significant. Scatter plot ( j ), Median tail moment in n=3 biological experiments, with indicated mice. Significance, unpaired two-tailed t-test, **P ≤ 0.01; ns, non-significant. Throughout the figure, WT denotes non-auxin-treated BARD1 AID/AID cells. For BARD1 Δ / Δ cells, BARD1 AID/AID cell lines were treated with doxycycline (2 μg/ml, 24 h) before addition of auxin (IAA, 1 mM) or dimethyl sulfoxide (DMSO) (carrier control), followed by olaparib.

Article Snippet: One hour after IAA addition, olaparib (S1060, Selleckchem) or cisplatin (P4394, Merck) was added to the indicated final concentrations.

Techniques: Viability Assay, Immunofluorescence, Imaging, Single Cell Gel Electrophoresis, MANN-WHITNEY, Two Tailed Test, Control

Journal: bioRxiv

Article Title: ATM safeguards DNA replication at endogenous base lesions

doi: 10.64898/2026.03.20.713105

Figure Lengend Snippet: a,b, Alkaline comet assays of indicated HCT-116 cell lines cultured with 50 µM β-mercaptoethanol or 250 µM N-acetyl cysteine (NAC) for 72-96 h ( a ). Cells were cultured at ambient (20%) or physiological (3%) O 2 conditions. Scatter plot data ( a ), n=2 biological experiments (80–100 cells per experiment; each color denotes one replicate). Significance, two sided Kruskal-Wallis H test with Dunn’s correction for multiple comparisons. ****P ≤ 0.0001; ns, non-significant. c, Seven-day clonogenic survival of indicated BARD1 AID/AID HCT-116 cell lines grown in the presence of indicated doses of olaparib (PARPi) under ambient (20%) or physiological (3%) O 2 conditions. d-g, Resazurin survival assay of the indicated BARD1 AID/AID HCT-116 cell lines grown for 7 days in the presence indicated doses of olaparib under ambient (20%) or physiological (3%) O 2 conditions, n=3 biological experiments, mean ± s.d. In experiments involving BARD1 Δ / Δ cells, all the indicated BARD1 AID/AID cell lines were cultured in doxycycline (2 μg/ml) for 24 h before auxin (IAA, 1 mM) or dimethyl sulfoxide (DMSO) (carrier control), prior to olaparib addition. Throughout figure, WT denotes non-auxin-treated BARD1 AID/AID cells.

Article Snippet: One hour after IAA addition, olaparib (S1060, Selleckchem) or cisplatin (P4394, Merck) was added to the indicated final concentrations.

Techniques: Cell Culture, Clonogenic Cell Survival Assay, Control

Journal: SLAS discovery : advancing life sciences R & D

Article Title: HTS discovery of PARP1-HPF1 complex inhibitors in cancer

doi: 10.1016/j.slasd.2023.10.003

Figure Lengend Snippet: Development of Assay Method. (A) Demonstration of HPF1-effect on PAR chain length. PARP1 was incubated with p18mer DNA (lane 1) and was PARylated by addition of NAD + for 15 min, leading to a smear of high MW PARP1 (lane 2). In lane 3, PARP1 was incubated with HPF1 without the addition of NAD+. In lane 4, upon addition of NAD + , PARylation of PARP1 is observed to yield much shorter chains. Lanes 5 and 6 show the same experiment as lanes 3 and 4, respectively, using a different preparation of HPF1. (B) Left: Heat map of HTS primary assay FP data in a 384-well plate. Negative controls and non-inhibitor samples are blue. Positive controls and inhibited samples are red. Column 24 (odd rows): Primary assay + 200 μM NAD+ (no FP signal), (even rows): Primary assay – 200 μM NAD+ (high FP signal). Column 23 (odd rows): olaparib 200 nM 1:2 fold dilution (top to bottom), (even rows): benzamide 200 μM 1:2 fold dilution (top to bottom). Six random wells were blind-spiked with 200 μM benzamide and demonstrates that the assay performs well throughout the plate. Right: Dose-response graphs for olaparib (IC 50 : 17 nM) and benzamide (IC 50 : 14 μM). (C) A primary screen was performed to determine the 25 most potent compounds using the PARP1–HPF1 complex. Data points >200 μM are not shown for graphical clarity. Compounds with IC 50 < 25 μM (dotted line) were subjected to a second round of testing in triplicate.

Article Snippet: Olaparib and benzamide were obtained from Tocris and Sigma-Aldrich, respectively, and stock solutions were prepared in DMSO.

Techniques: Incubation

Journal: Cells

Article Title: Targeted PARP Inhibition Combined with FGFR1 Blockade is Synthetically Lethal to Malignant Cells in Patients with Pancreatic Cancer

doi: 10.3390/cells9040911

Figure Lengend Snippet: Enhanced FGFR1 and PARP1 expression characterizes FGFR1 inhibitor-resistant pancreatic cells at mRNA and protein levels. ( A ) Histograms of the FGFR1 (upper panel) and PARP1 (lower panel) RNAseq expression profile in dasatinib-sensitive PANC0403, PANC0504, and PANC1005 cells compared with dasatinib-resistant SU8686, MiaPaCa2, and PANC-1 pancreatic cancer cells in the GSE9357/GP10558/GDS5627 cohort. ( B ) Representative Western blot data showing the differential expression of FGFR1 and PARP in PANC-1, AsPC1, PANC0403, or SUIT-2 cells. ( C ) Line graphs showing the effect of 1–10 μM PD173074 or olaparib on the viability of PANC-1 (left panel) or SUIT-2 (right panel) cells. ( D ) Representative Western blot images of the effect of 4.5 μM PD173074 or 3.5 μM olaparib on the expression level of PARP or FGFR1 protein in the PANC-1 and SUIT-2 cells. β-actin served as the loading control.

Article Snippet: Sections from tumor samples resected from the PD173074 and/or olaparib-treated tumor-bearing BALB/c mice were stained using primary antibodies against FGFR1 (#9740; 1:400, Cell Signaling Technology Inc., Danvers, MA, USA), cleaved PARP (#5625; 1:400, Cell Signaling Technology), Ki-67 (#ab15580; 1:500, Abcam, Cambridge, MA, USA), and Bcl-2 (#15071; 1:400, Cell Signaling Technology).

Techniques: Expressing, Western Blot, Quantitative Proteomics, Control

Journal: Cells

Article Title: Targeted PARP Inhibition Combined with FGFR1 Blockade is Synthetically Lethal to Malignant Cells in Patients with Pancreatic Cancer

doi: 10.3390/cells9040911

Figure Lengend Snippet: Olaparib-induced pharmacological inhibition of PARP1 synergistically enhances the therapeutic effect of PD173074. ( A ) Graphical representation of changes in PANC-1 and SUIT-2 cell viability when treated with 4.5 μM PD173074 and/or 3.5 μM olaparib for 48 h. ( B ) Effect of combining 2.5–10 μM PD173074 and 0.5–5 μM olaparib on the viability of PANC-1 cells. ( C ) Isobologram and drug combination index plot indicating synergy between different concentrations of PD173074 and olaparib. ( D ) The effect of treatment with 4.5 μM PD173074 and/or 3.5 μM olaparib on the expression level of PARP, cleaved PARP, pro-caspase-9, cleaved-caspase-9, pro-caspase-3, cleaved-caspase-3, and Bcl-xL proteins as shown by Western blot analysis. * p < 0.05, ** p < 0.01, (PANC-1: treated vs. CTL); * p < 0.05, ** p < 0.01 (SUIT-2: treated vs. CTL); CTL, control; Combo, PD173074 + olaparib combination therapy; Fa, fraction affected; CI, combination index; DRI, dose-reduction index.

Article Snippet: Sections from tumor samples resected from the PD173074 and/or olaparib-treated tumor-bearing BALB/c mice were stained using primary antibodies against FGFR1 (#9740; 1:400, Cell Signaling Technology Inc., Danvers, MA, USA), cleaved PARP (#5625; 1:400, Cell Signaling Technology), Ki-67 (#ab15580; 1:500, Abcam, Cambridge, MA, USA), and Bcl-2 (#15071; 1:400, Cell Signaling Technology).

Techniques: Inhibition, Expressing, Western Blot, Control

Journal: Cells

Article Title: Targeted PARP Inhibition Combined with FGFR1 Blockade is Synthetically Lethal to Malignant Cells in Patients with Pancreatic Cancer

doi: 10.3390/cells9040911

Figure Lengend Snippet: Olaparib alone or in synergism with PD173074 suppresses the oncogenic cancer stem cell-like phenotype of PDAC cells through an apoptosis-related impairment of DNA repair. ( A ) Tumorsphere formation assay images and histograms showing the effect of PD173074 and/or olaparib on the tumorsphere formation efficiency of the PANC-1 or SUIT-2 cells. ( B ) Representative colony formation assay data (left) and graph (right) showing the effect of PD17374 and/or olaparib on the number of colonies formed by PANC-1 or SUIT-2 cells. ( C ) Immunofluorescence images and histograms of γ-H2AX (red), RAD51 (green), and nuclear DAPI (blue) staining in PANC-1 cells treated with PD173074 and/or olaparib compared with the control. Scale bar, 10 μm. ( D ) Downregulation of FGFR1, Sox2, and Nanog protein expression levels in PD173074-treated Panc-1 cells as determined by Western blot analysis. β-actin was used as a loading control. * p < 0.05

Article Snippet: Sections from tumor samples resected from the PD173074 and/or olaparib-treated tumor-bearing BALB/c mice were stained using primary antibodies against FGFR1 (#9740; 1:400, Cell Signaling Technology Inc., Danvers, MA, USA), cleaved PARP (#5625; 1:400, Cell Signaling Technology), Ki-67 (#ab15580; 1:500, Abcam, Cambridge, MA, USA), and Bcl-2 (#15071; 1:400, Cell Signaling Technology).

Techniques: Tube Formation Assay, Colony Assay, Immunofluorescence, Staining, Control, Expressing, Western Blot

Journal: Cells

Article Title: Targeted PARP Inhibition Combined with FGFR1 Blockade is Synthetically Lethal to Malignant Cells in Patients with Pancreatic Cancer

doi: 10.3390/cells9040911

Figure Lengend Snippet: In vivo, PDAC cancer stem cells are more sensitive to PD173074 in the absence of PARP1. ( A ) Experimental chart of in vivo studies. Line graphs of the differential effect of ( B ) early or ( C ) late initiation of treatment with PD173074 and/or olaparib on BALB/c mice bearing PANC-1 tumorsphere-derived tumors. Graphical representation of tumor weight at the end of the experiment for ( D ) early and ( E ) late treatment initiation groups. ( F ) Frequency of tumor growth and correlative statistical analysis of treatment regimen in the PD173074- and/or olaparib-treated mice. ( G ) Representative immunohistochemical staining images showing differential expression of cleaved PARP, Ki-67, FGFR1, and Bcl-2 in tumors extracted from the PD173074- and/or olaparib-treated NOD/SCID mice. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Sections from tumor samples resected from the PD173074 and/or olaparib-treated tumor-bearing BALB/c mice were stained using primary antibodies against FGFR1 (#9740; 1:400, Cell Signaling Technology Inc., Danvers, MA, USA), cleaved PARP (#5625; 1:400, Cell Signaling Technology), Ki-67 (#ab15580; 1:500, Abcam, Cambridge, MA, USA), and Bcl-2 (#15071; 1:400, Cell Signaling Technology).

Techniques: In Vivo, Derivative Assay, Immunohistochemical staining, Staining, Quantitative Proteomics