Top

Breast Cancer Research and Treatment

Published in:

30-10-2023 | Breast Cancer | Preclinical study

Olaparib enhances sensitization of BRCA-proficient breast cancer cells to x-rays and protons

Authors: Sohee Park, Changhoon Choi, Haeyoung Kim, Yong Jae Shin, Yunjeong Oh, Won Park, Won Kyung Cho, Nalee Kim

Published in: Breast Cancer Research and Treatment | Issue 3/2024

Abstract

Purpose

This study aimed to compare the radiosensitizing effect of the PARP inhibitor, Olaparib, between proton and X-rays irradiations in BRCA-proficient breast cancer (BC) cells.

Methods

Two BRCA-proficient BC cell lines, MDA-MB-231 and T47D BC, were used. Cell proliferation was assessed using the CCK-8 assay, and radiosensitivity was determined through the clonogenic survival assay. Flow cytometry was employed to analyze cell cycle distribution and apoptosis. The kinetics of DNA damage repair were evaluated using γH2AX immunofluorescence imaging and the comet assay. Tumor spheroid assays were conducted to test radiosensitivity in a three-dimensional culture condition.

Results

Olaparib sensitized both MDA-MB-231 and T47D cells to proton and X-ray irradiation in the clonogenic assay. MDA-MB-231 cells exhibited a higher dose enhancement factor for Olaparib than T47D cells. Olaparib increased radiation-induced G2/M cell cycle arrest and apoptosis specifically in MDA-MB-231 cells. γH2AX immunostaining and the comet assay showed Olaparib augmented radiation-induced DNA damage and apoptosis. The enhancement effect of Olaparib was more pronounced in proton irradiation than in X-ray irradiation, particularly in MDA-MB-231 cells than T47D cells. Both radiation and Olaparib dose-dependently inhibited spheroid growth in both cell lines. The synergy scores demonstrated that Olaparib interacted more strongly with protons than X-rays. The addition of an ATR inhibitor further enhanced Olaparib-induced proton radiosensitization in MDA-MB-231 cells.

Conclusion

This study found that Olaparib enhanced radiation efficacy in BRCA-proficient breast cancer cells, with a more pronounced effect observed with proton irradiation compared to X-ray irradiation. Combining Olaparib with an ATR inhibitor increased the radiosensitizing effect of protons.

Kowalchuk RO, Corbin KS, Jimenez RB (2022) Particle therapy for breast cancer. Cancers (Basel). https://doi.org/10.3390/cancers14041066CrossRefPubMed

Baumann BC, Mitra N, Harton JG, Xiao Y, Wojcieszynski AP, Gabriel PE, Zhong H, Geng H, Doucette A, Wei J, O’Dwyer PJ, Bekelman JE, Metz JM (2020) Comparative effectiveness of proton vs photon therapy as part of concurrent chemoradiotherapy for locally advanced cancer. JAMA Oncol 6:237–246. https://doi.org/10.1001/jamaoncol.2019.4889CrossRefPubMed

Tommasino F, Durante M (2015) Proton radiobiology. Cancers (Basel) 7:353–381. https://doi.org/10.3390/cancers7010353CrossRefPubMed

Sorensen BS, Pawelke J, Bauer J, Burnet NG, Dasu A, Hoyer M, Karger CP, Krause M, Schwarz M, Underwood TSA, Wagenaar D, Whitfield GA, Luhr A (2021) Does the uncertainty in relative biological effectiveness affect patient treatment in proton therapy? Radiother Oncol 163:177–184. https://doi.org/10.1016/j.radonc.2021.08.016CrossRefPubMed

Grosse N, Fontana AO, Hug EB, Lomax A, Coray A, Augsburger M, Paganetti H, Sartori AA, Pruschy M (2014) Deficiency in homologous recombination renders Mammalian cells more sensitive to proton versus photon irradiation. Int J Radiat Oncol Biol Phys 88:175–181. https://doi.org/10.1016/j.ijrobp.2013.09.041CrossRefPubMed

Fontana AO, Augsburger MA, Grosse N, Guckenberger M, Lomax AJ, Sartori AA, Pruschy MN (2015) Differential DNA repair pathway choice in cancer cells after proton- and photon-irradiation. Radiother Oncol 116:374–380. https://doi.org/10.1016/j.radonc.2015.08.014CrossRefPubMed

Bright SJ, Flint DB, Chakraborty S, McFadden CH, Yoon DS, Bronk L, Titt U, Mohan R, Grosshans DR, Sumazin P, Shaitelman SF, Asaithamby A, Sawakuchi GO (2019) Nonhomologous end joining is more important than proton linear energy transfer in dictating cell death. Int J Radiat Oncol Biol Phys 105:1119–1125. https://doi.org/10.1016/j.ijrobp.2019.08.011CrossRefPubMedPubMedCentral

Bai P (2015) Biology of poly(ADP-ribose) polymerases: the factotums of cell maintenance. Mol Cell 58:947–958. https://doi.org/10.1016/j.molcel.2015.01.034CrossRefPubMed

Zheng F, Zhang Y, Chen S, Weng X, Rao Y, Fang H (2020) Mechanism and current progress of poly ADP-ribose polymerase (PARP) inhibitors in the treatment of ovarian cancer. Biomed Pharmacother 123:109661. https://doi.org/10.1016/j.biopha.2019.109661CrossRefPubMed

10.

Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, Kyle S, Meuth M, Curtin NJ, Helleday T (2005) Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 434:913–917. https://doi.org/10.1038/nature03443CrossRefPubMed

11.

Pommier Y, O’Connor MJ, de Bono J (2016) Laying a trap to kill cancer cells: PARP inhibitors and their mechanisms of action. Sci Transl Med. https://doi.org/10.1126/scitranslmed.aaf9246CrossRefPubMedPubMedCentral

12.

Cortesi L, Rugo HS, Jackisch C (2021) An overview of PARP inhibitors for the treatment of breast cancer. Target Oncol 16:255–282. https://doi.org/10.1007/s11523-021-00796-4CrossRefPubMedPubMedCentral

13.

Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA, Richardson TB, Santarosa M, Dillon KJ, Hickson I, Knights C, Martin NM, Jackson SP, Smith GC, Ashworth A (2005) Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 434:917–921. https://doi.org/10.1038/nature03445CrossRefPubMed

14.

Robson M, Goessl C, Domchek S (2017) Olaparib for metastatic germline BRCA-mutated breast cancer. N Engl J Med 377:1792–1793. https://doi.org/10.1056/NEJMc1711644CrossRefPubMed

15.

Litton JK, Rugo HS, Ettl J, Hurvitz SA, Goncalves A, Lee KH, Fehrenbacher L, Yerushalmi R, Mina LA, Martin M, Roche H, Im YH, Quek RGW, Markova D, Tudor IC, Hannah AL, Eiermann W, Blum JL (2018) Talazoparib in patients with advanced breast cancer and a germline BRCA mutation. N Engl J Med 379:753–763. https://doi.org/10.1056/NEJMoa1802905CrossRefPubMedPubMedCentral

16.

Barcellini A, Loap P, Murata K, Villa R, Kirova Y, Okonogi N, Orlandi E (2021) PARP inhibitors in combination with radiotherapy: to do or not to do? Cancers (Basel). https://doi.org/10.3390/cancers13215380CrossRefPubMed

17.

Cesaire M, Thariat J, Candeias SM, Stefan D, Saintigny Y, Chevalier F (2018) Combining PARP inhibition, radiation, and immunotherapy: a possible strategy to improve the treatment of cancer? Int J Mol Sci. https://doi.org/10.3390/ijms19123793CrossRefPubMedPubMedCentral

18.

Lakomy DS, Urbauer DL, Westin SN, Lin LL (2020) Phase I study of the PARP inhibitor talazoparib with radiation therapy for locally recurrent gynecologic cancers. Clin Transl Radiat Oncol 21:56–61. https://doi.org/10.1016/j.ctro.2019.12.005CrossRefPubMed

19.

Tutt ANJ, Garber JE, Kaufman B, Viale G, Fumagalli D, Rastogi P, Gelber RD, de Azambuja E, Fielding A, Balmana J, Domchek SM, Gelmon KA, Hollingsworth SJ, Korde LA, Linderholm B, Bandos H, Senkus E, Suga JM, Shao Z, Pippas AW, Nowecki Z, Huzarski T, Ganz PA, Lucas PC, Baker N, Loibl S, McConnell R, Piccart M, Schmutzler R, Steger GG, Costantino JP, Arahmani A, Wolmark N, McFadden E, Karantza V, Lakhani SR, Yothers G, Campbell C, Geyer CE Jr, Olympi ACTSC, Investigators (2021) Adjuvant olaparib for patients with BRCA1- or BRCA2-mutated breast cancer. N Engl J Med 384:2394–2405. https://doi.org/10.1056/NEJMoa2105215CrossRefPubMedPubMedCentral

20.

Robson M, Im SA, Senkus E, Xu B, Domchek SM, Masuda N, Delaloge S, Li W, Tung N, Armstrong A, Wu W, Goessl C, Runswick S, Conte P (2017) Olaparib for metastatic breast cancer in patients with a germline BRCA mutation. N Engl J Med 377:523–533. https://doi.org/10.1056/NEJMoa1706450CrossRefPubMed

21.

Keung MY, Wu Y, Badar F, Vadgama JV (2020) Response of breast cancer cells to PARP inhibitors is independent of BRCA status. J Clin Med. https://doi.org/10.3390/jcm9040940CrossRefPubMedPubMedCentral

22.

Dungey FA, Loser DA, Chalmers AJ (2008) Replication-dependent radiosensitization of human glioma cells by inhibition of poly(ADP-ribose) polymerase: mechanisms and therapeutic potential. Int J Radiat Oncol Biol Phys 72:1188–1197. https://doi.org/10.1016/j.ijrobp.2008.07.031CrossRefPubMed

23.

Wang N, Yang Y, Jin D, Zhang Z, Shen K, Yang J, Chen H, Zhao X, Yang L, Lu H (2022) PARP inhibitor resistance in breast and gynecological cancer: resistance mechanisms and combination therapy strategies. Front Pharmacol 13:967633. https://doi.org/10.3389/fphar.2022.967633CrossRefPubMedPubMedCentral

24.

Ianevski A, He L, Aittokallio T, Tang J (2017) SynergyFinder: a web application for analyzing drug combination dose-response matrix data. Bioinformatics 33:2413–2415. https://doi.org/10.1093/bioinformatics/btx162CrossRefPubMedPubMedCentral

25.

Parsels LA, Engelke CG, Parsels J, Flanagan SA, Zhang Q, Tanska D, Wahl DR, Canman CE, Lawrence TS, Morgan MA (2021) Combinatorial efficacy of olaparib with radiation and ATR inhibitor requires PARP1 protein in homologous recombination-proficient pancreatic cancer. Mol Cancer Ther 20:263–273. https://doi.org/10.1158/1535-7163.MCT-20-0365CrossRefPubMed

26.

Hirai T, Saito S, Fujimori H, Matsushita K, Nishio T, Okayasu R, Masutani M (2016) Radiosensitization by PARP inhibition to proton beam irradiation in cancer cells. Biochem Biophys Res Commun 478:234–240. https://doi.org/10.1016/j.bbrc.2016.07.062CrossRefPubMed

27.

Kageyama SI, Junyan D, Hojo H, Motegi A, Nakamura M, Tsuchihara K, Akimoto T (2020) PARP inhibitor olaparib sensitizes esophageal carcinoma cells to fractionated proton irradiation. J Radiat Res 61:177–186. https://doi.org/10.1093/jrr/rrz088CrossRefPubMedPubMedCentral

28.

Wang L, Cao J, Wang X, Lin E, Wang Z, Li Y, Li Y, Chen M, Wang X, Jiang B, Zhang R, Sahoo N, Zhang X, Zhu XR, Myers JN, Frank SJ (2020) Proton and photon radiosensitization effects of niraparib, a PARP-1/-2 inhibitor, on human head and neck cancer cells. Head Neck 42:2244–2256. https://doi.org/10.1002/hed.26155CrossRefPubMed

29.

Lee DW, Kim JE, Lee GH, Son A, Park HC, Oh D, Jo K, Choi C (2022) High-throughput 3D tumor spheroid array platform for evaluating sensitivity of proton-drug combinations. Int J Mol Sci. https://doi.org/10.3390/ijms23020587CrossRefPubMedPubMedCentral

30.

Zhou C, Fabbrizi MR, Hughes JR, Grundy GJ, Parsons JL (2022) Effectiveness of PARP inhibition in enhancing the radiosensitivity of 3D spheroids of head and neck squamous cell carcinoma. Front Oncol 12:940377. https://doi.org/10.3389/fonc.2022.940377CrossRefPubMedPubMedCentral

31.

Elstrodt F, Hollestelle A, Nagel JH, Gorin M, Wasielewski M, van den Ouweland A, Merajver SD, Ethier SP, Schutte M (2006) BRCA1 mutation analysis of 41 human breast cancer cell lines reveals three new deleterious mutants. Cancer Res 66:41–45. https://doi.org/10.1158/0008-5472.CAN-05-2853CrossRefPubMed

32.

Zhen Y, Zhang Y, Yu Y (2017) A cell-line-specific atlas of PARP-mediated protein Asp/Glu-ADP-ribosylation in breast cancer. Cell Rep 21:2326–2337. https://doi.org/10.1016/j.celrep.2017.10.106CrossRefPubMedPubMedCentral

33.

Min A, Im SA, Kim DK, Song SH, Kim HJ, Lee KH, Kim TY, Han SW, Oh DY, Kim TY, O’Connor MJ, Bang YJ (2015) Histone deacetylase inhibitor, suberoylanilide hydroxamic acid (SAHA), enhances anti-tumor effects of the poly (ADP-ribose) polymerase (PARP) inhibitor olaparib in triple-negative breast cancer cells. Breast Cancer Res 17:33. https://doi.org/10.1186/s13058-015-0534-yCrossRefPubMedPubMedCentral

34.

Berardi D, Hunter Y, van den Driest L, Farrell G, Rattray NJW, Rattray Z (2022) The differential metabolic signature of breast cancer cellular response to olaparib treatment. Cancers (Basel). https://doi.org/10.3390/cancers14153661CrossRefPubMed

35.

Waissi W, Nicol A, Jung M, Rousseau M, Jarnet D, Noel G, Burckel H (2021) Radiosensitizing pancreatic cancer with PARP inhibitor and gemcitabine: an in vivo and a whole-transcriptome analysis after proton or photon irradiation. Cancers (Basel). https://doi.org/10.3390/cancers13030527CrossRefPubMed

36.

Wera AC, Lobbens A, Stoyanov M, Lucas S, Michiels C (2019) Radiation-induced synthetic lethality: combination of poly(ADP-ribose) polymerase and RAD51 inhibitors to sensitize cells to proton irradiation. Cell Cycle 18:1770–1783. https://doi.org/10.1080/15384101.2019.1632640CrossRefPubMedPubMedCentral

37.

Huart C, Fransolet M, Demazy C, Le Calve B, Lucas S, Michiels C, Wera AC (2022) Taking advantage of the senescence-promoting effect of olaparib after X-ray and proton irradiation using the senolytic drug, ABT-263. Cancers (Basel). https://doi.org/10.3390/cancers14061460CrossRefPubMed

38.

Bright SJ, Flint DB, Martinus DKJ, Turner BX, Manandhar M, Kacem MB, McFadden CH, Yap TA, Shaitelman SF, Sawakuchi GO (2022) Targeted inhibition of DNA-PKcs, ATM, ATR, PARP, and Rad51 modulate response to X rays and protons. Radiat Res 198:336–346. https://doi.org/10.1667/RADE-22-00040.1CrossRefPubMedPubMedCentral

39.

Vitti ET, Kacperek A, Parsons JL (2020) Targeting DNA double-strand break repair enhances radiosensitivity of HPV-positive and HPV-negative head and neck squamous cell carcinoma to photons and protons. Cancers (Basel). https://doi.org/10.3390/cancers12061490CrossRefPubMed

40.

Yazinski SA, Comaills V, Buisson R, Genois MM, Nguyen HD, Ho CK, Todorova Kwan T, Morris R, Lauffer S, Nussenzweig A, Ramaswamy S, Benes CH, Haber DA, Maheswaran S, Birrer MJ, Zou L (2017) ATR inhibition disrupts rewired homologous recombination and fork protection pathways in PARP inhibitor-resistant BRCA-deficient cancer cells. Genes Dev 31:318–332. https://doi.org/10.1101/gad.290957.116CrossRefPubMedPubMedCentral

41.

Gralewska P, Gajek A, Marczak A, Mikula M, Ostrowski J, Sliwinska A, Rogalska A (2020) PARP inhibition increases the reliance on ATR/CHK1 checkpoint signaling leading to synthetic lethality-an alternative treatment strategy for epithelial ovarian cancer cells independent from HR effectiveness. Int J Mol Sci. https://doi.org/10.3390/ijms21249715CrossRefPubMedPubMedCentral

42.

Price JM, Prabhakaran A, West CML (2023) Predicting tumour radiosensitivity to deliver precision radiotherapy. Nat Rev Clin Oncol 20:83–98. https://doi.org/10.1038/s41571-022-00709-yCrossRefPubMed

43.

Zhang J, Si J, Gan L, Zhou R, Guo M, Zhang H (2021) Harnessing the targeting potential of differential radiobiological effects of photon versus particle radiation for cancer treatment. J Cell Physiol 236:1695–1711. https://doi.org/10.1002/jcp.29960CrossRefPubMed

44.

Choi C, Cho WK, Park S, Shin SW, Park W, Kim H, Choi DH (2020) Checkpoint kinase 1 (CHK1) inhibition enhances the sensitivity of triple-negative breast cancer cells to proton irradiation via Rad51 downregulation. Int J Mol Sci. https://doi.org/10.3390/ijms21082691CrossRefPubMedPubMedCentral

45.

Choi C, Park S, Cho WK, Choi DH (2019) Cyclin D1 is associated with radiosensitivity of triple-negative breast cancer cells to proton beam irradiation. Int J Mol Sci. https://doi.org/10.3390/ijms20194943CrossRefPubMedPubMedCentral

Title: Olaparib enhances sensitization of BRCA-proficient breast cancer cells to x-rays and protons
Authors: Sohee Park
Changhoon Choi
Haeyoung Kim
Yong Jae Shin
Yunjeong Oh
Won Park
Won Kyung Cho
Nalee Kim
Publication date: 30-10-2023
Publisher: Springer US
Keywords: Breast Cancer
Breast Cancer
Published in: Breast Cancer Research and Treatment / Issue 3/2024
Print ISSN: 0167-6806
Electronic ISSN: 1573-7217
DOI: https://doi.org/10.1007/s10549-023-07150-4

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 3/2024

Association of physical weight statuses defined by body mass index (BMI) with molecular subtypes of premenopausal breast cancer: a systematic review and meta-analysis

Assessment of breast cancer chemotherapy dose reduction in an integrated healthcare delivery system

Secretory breast carcinoma: clinicopathological features and prognosis of 52 patients

Nodal pCR and overall survival following neoadjuvant chemotherapy for node positive ER+/Her2- breast cancer

Interleukin-6 and C-Reactive protein in metastatic breast cancer patients treated with eribulin

Screening mammographic performance by race and age in the National Mammography Database: 29,479,665 screening mammograms from 13,181,241 women

Keynote webinar | Spotlight on antibody–drug conjugates in cancer