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Olaparib

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Olaparib is a poly (ADP-ribose) polymerase (PARP) inhibitor. It is a small molecule that binds to and inhibits the PARP enzyme, which is involved in DNA repair processes within cells.

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Lab products found in correlation

Rpmi medium, by Merck Group (2 mentions) Mk 1775, by Abcam (2 mentions) Azd 7762, by Abcam (2 mentions) Azd4547, by Abcam (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Lapatinib, by Abcam (2 mentions) Dmem medium, by Thermo Fisher Scientific (2 mentions) Topotecan, by Cayman Chemical (2 mentions) Enasidenib, by Abcam (2 mentions) D2hg assay kit, by Abcam (2 mentions) 2 hydroxypropyl β cyclodextrin, by Merck Group (2 mentions) Olaparib, by Selleck Chemicals (2 mentions) Hydroxyurea, by Merck Group (1 mentions) Camptothecin, by Merck Group (1 mentions) Ly2606368, by Selleck Chemicals (1 mentions) Mitomycin c, by Merck Group (1 mentions) Etoposide, by Merck Group (1 mentions) Cisplatin, by Merck Group (1 mentions) Ku55933, by Selleck Chemicals (1 mentions) Azd6738, by Selleck Chemicals (1 mentions)

6 protocols using olaparib

1

Evaluating Small Molecule Inhibitors

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KU-55933, AZD6738, and LY2606368 were purchased from Selleck Chemicals. Olaparib was purchased from BioVision Inc. Hydroxyurea, cisplatin, mitomycin C (MMC), camptothecin, and etoposide were purchased from Sigma-Aldrich.
Su D., Feng X., Colic M., Wang Y., Zhang C., Wang C., Tang M., Hart T, & Chen J. (2020). CRISPR/CAS9-based DNA damage response screens reveal gene-drug interactions. DNA repair, 87, 102803.
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2

Establishment and Characterization of EAC Cell Lines

Cited 1 time
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The primary cell line panel was derived from EAC cases included in the ICGC sequencing study, including MFD (Tim Underwood, Southampton, OCCAMS consortium member), OES127 (Anna Grabowska, Nottingham, OCCAMS consortium member) and CAM02 (organoid, Mathew Garnett, Cambridge). The MFD line required 10% fetal calf serum (PAA) in DMEM medium (Invitrogen, ThermoFisher Scientific) and the CAM02 culture method was as previously described [51 (link)]. The feeder layer system was used to expand OES127 lines. The established EAC lines, SK-GT-4, OAC-P4C, OACM5.1C, and OE33 were cultured in RPMI medium (Sigma) with 10% fetal calf serum, except for FLO-1, which was grown in DMEM with 10% fetal calf serum. The identity of all cell lines was verified by short tandem repeat (STR) profiling and routinely examined for mycoplasma contamination.
Small molecular inhibitors used for treatment were: Lapatinib, AZD-4547, Olaparib, MK-1775 and AZD-7762 (BioVision), Crizotinib (LKT Labs) and Topotecan (Cayman Chemical). Inhibitors were diluted to working concentrations in DMSO (Sigma).
Secrier M., Li X., de Silva N., Eldridge M.D., Contino G., Bornschein J., MacRae S., Grehan N., O’Donovan M., Miremadi A., Yang T.P., Bower L., Chettouh H., Crawte J., Galeano-Dalmau N., Grabowska A., Saunders J., Underwood T., Waddell N., Barbour A.P., Nutzinger B., Achilleos A., Edwards P.A., Lynch A.G., Tavaré S, & Fitzgerald R.C. (2016). Mutational signatures in esophageal adenocarcinoma define etiologically distinct subgroups with therapeutic relevance. Nature genetics, 48(10), 1131-1141.
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3

Olaparib Sensitivity in RPE1-hTERT Cells

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About 2 × 103 cells each for indicated Lenti-sgRNA transfected RPE1-hTERT TP53−/− BRCA1−/− cells used in the study were seeded onto 6-well plates in triplicate, treated with various doses (0, 50, and 100 nM) of Olaparib (BioVision), and then incubated for 14 days. Colonies were fixed and stained with 0.5% crystal violet. Relative cell viability was measured using ImageJ (National Institutes of Health).
Tang M., Chen Z., Wang C., Feng X., Lee N., Huang M., Zhang H., Li S., Xiong Y, & Chen J. (2022). Histone chaperone ASF1 acts with RIF1 to promote DNA end joining in BRCA1-deficient cells. The Journal of Biological Chemistry, 298(6), 101979.
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4

Establishment and Characterization of EAC Cell Lines

Cited 1 time
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The primary cell line panel was derived from EAC cases included in the ICGC sequencing study, including MFD (Tim Underwood, Southampton, OCCAMS consortium member), OES127 (Anna Grabowska, Nottingham, OCCAMS consortium member) and CAM02 (organoid, Mathew Garnett, Cambridge). The MFD line required 10% fetal calf serum (PAA) in DMEM medium (Invitrogen, ThermoFisher Scientific) and the CAM02 culture method was as previously described [51 (link)]. The feeder layer system was used to expand OES127 lines. The established EAC lines, SK-GT-4, OAC-P4C, OACM5.1C, and OE33 were cultured in RPMI medium (Sigma) with 10% fetal calf serum, except for FLO-1, which was grown in DMEM with 10% fetal calf serum. The identity of all cell lines was verified by short tandem repeat (STR) profiling and routinely examined for mycoplasma contamination.
Small molecular inhibitors used for treatment were: Lapatinib, AZD-4547, Olaparib, MK-1775 and AZD-7762 (BioVision), Crizotinib (LKT Labs) and Topotecan (Cayman Chemical). Inhibitors were diluted to working concentrations in DMSO (Sigma).
Secrier M., Li X., de Silva N., Eldridge M.D., Contino G., Bornschein J., MacRae S., Grehan N., O’Donovan M., Miremadi A., Yang T.P., Bower L., Chettouh H., Crawte J., Galeano-Dalmau N., Grabowska A., Saunders J., Underwood T., Waddell N., Barbour A.P., Nutzinger B., Achilleos A., Edwards P.A., Lynch A.G., Tavaré S, & Fitzgerald R.C. (2016). Mutational signatures in esophageal adenocarcinoma define etiologically distinct subgroups with therapeutic relevance. Nature genetics, 48(10), 1131-1141.
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5

In vivo Olaparib and Enasidenib Pharmacokinetics

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Olaparib (AZD2281) (Selleckchem, Cat#S1060) was dissolved in PBS containing 10% (w/v) 2-hydroxy-propyl-beta-cyclodextrin (Sigma) at a concentration of 20mg/ml. For in vivo studies, Olaparib was administered IP at 100 mg/kg once daily. Enasidenib was purchased from LeadGen Labs, USA and dissolved in 0.5% methylcellulose and 0.2% Tween 80 in PBS at a concentration of 4 mg/ml and administered via oral gavage at 40 mg/kg once daily. 2-HG was measured in plasma before and after treatment with vehicle, Enasidenib, or Olaparib in triplicates with the D-2-Hydroxyglutarate (D2HG) assay kit (Biovision, Cat#K970–100, Sigma, Cat# MAK320–1KT) according to manufacturer’s protocol.
Gbyli R., Song Y., Liu W., Gao Y., Biancon G., Chandhok N.S., Wang X., Fu X., Patel A., Sundaram R., Tebaldi T., Mamillapalli P., Zeidan A.M., Flavell R.A., Prebet T., Bindra R.S, & Halene S. (2022). In Vivo Anti-Tumor Effect of PARP Inhibition in IDH1/2 Mutant MDS/AML Resistant to Targeted Inhibitors of Mutant IDH1/2. Leukemia, 36(5), 1313-1323.
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6

In vivo Olaparib and Enasidenib Pharmacokinetics

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Olaparib (AZD2281) (Selleckchem, Cat#S1060) was dissolved in PBS containing 10% (w/v) 2-hydroxy-propyl-beta-cyclodextrin (Sigma) at a concentration of 20mg/ml. For in vivo studies, Olaparib was administered IP at 100 mg/kg once daily. Enasidenib was purchased from LeadGen Labs, USA and dissolved in 0.5% methylcellulose and 0.2% Tween 80 in PBS at a concentration of 4 mg/ml and administered via oral gavage at 40 mg/kg once daily. 2-HG was measured in plasma before and after treatment with vehicle, Enasidenib, or Olaparib in triplicates with the D-2-Hydroxyglutarate (D2HG) assay kit (Biovision, Cat#K970–100, Sigma, Cat# MAK320–1KT) according to manufacturer’s protocol.
Gbyli R., Song Y., Liu W., Gao Y., Biancon G., Chandhok N.S., Wang X., Fu X., Patel A., Sundaram R., Tebaldi T., Mamillapalli P., Zeidan A.M., Flavell R.A., Prebet T., Bindra R.S, & Halene S. (2022). In Vivo Anti-Tumor Effect of PARP Inhibition in IDH1/2 Mutant MDS/AML Resistant to Targeted Inhibitors of Mutant IDH1/2. Leukemia, 36(5), 1313-1323.
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