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Cited by 20PARP inhibition in cancer therapyEstrogen and related hormone effectsAdvanced Breast Cancer Therapies

Estrogen therapy induces receptor-dependent DNA damage enhanced by PARP inhibition in ER+ breast cancer

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Traphagen, Nicole A., Schwartz, Gary N., Tau, Steven, Roberts, Alyssa M., Jiang, Amanda, Hosford, Sarah R., Marotti, Jonathan D., Goen, Abigail E. +9 more·2023·Clinical Cancer Research
DOI 10.1158/1078-0432.ccr-23-0488PMC PMC10528687

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Abstract

Purpose: Clinical evidence indicates that treatment with estrogens elicits anticancer effects in ∼30% of patients with advanced endocrine-resistant estrogen receptor α (ER)-positive breast cancer. Despite the proven efficacy of estrogen therapy, its mechanism of action is unclear and this treatment remains underused. Mechanistic understanding may offer strategies to enhance therapeutic efficacy. Experimental Design: We performed genome-wide CRISPR/Cas9 screening and transcriptomic profiling in long-term estrogen-deprived ER+ breast cancer cells to identify pathways required for therapeutic response to the estrogen 17β-estradiol (E2). We validated findings in cell lines, patient-derived xenografts (PDX), and patient samples, and developed a novel combination treatment through testing in cell lines and PDX models. Results: Cells treated with E2 exhibited replication-dependent markers of DNA damage and the DNA damage response prior to apoptosis. Such DNA damage was partially driven by the formation of DNA:RNA hybrids (R-loops). Pharmacologic suppression of the DNA damage response via PARP inhibition with olaparib enhanced E2-induced DNA damage. PARP inhibition synergized with E2 to suppress growth and prevent tumor recurrence in BRCA1/2-mutant and BRCA1/2-wild-type cell line and PDX models. Conclusions: E2-induced ER activity drives DNA damage and growth inhibition in endocrine-resistant breast cancer cells. Inhibition of the DNA damage response using drugs such as PARP inhibitors can enhance therapeutic response to E2. These findings warrant clinical exploration of the combination of E2 with DNA damage response inhibitors in advanced ER+ breast cancer, and suggest that PARP inhibitors may synergize with therapeutics that exacerbate transcriptional stress.

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  • eGR is an unbiased metric of in vivo antiproliferative drug effect.

    “Our method is based on eGR, an unbiased metric of in vivo antiproliferative drug effect (30), which is similar to two unbiased metrics of in vitro antiproliferative drug effect, the DIP rate proposed by Harris and colleagues (33), and the drug-induced GR inhibition by Hafner and colleagues (32), and it offers an easy algebraic way for calculating GR for in vivo studies.”

    From Statistical Assessment of Drug Synergy from <i>In Vivo</i> Combination Studies Using Mouse Tumor Models (2023) · cites this paper as [30]

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  1. [1]Shiino S, Kinoshita T, Yoshida M, Jimbo K, Asaga S, Takayama S, Prognostic Impact of Discordance in Hormone Receptor Status Between Primary and Recurrent Sites in Patients With Recurrent Breast Cancer. Clinical breast cancer 2016; 16(4): e133– 40. 27268749
  2. [2]Osborne CK, Pippen J, Jones SE, Parker LM, Ellis M, Come S, Double-blind, randomized trial comparing the efficacy and tolerability of fulvestrant versus anastrozole in postmenopausal women with advanced breast cancer progressing on prior endocrine therapy: results of a North American trial. J Clin Oncol 2002; 20(16): 3386– 95. 12177098
  3. [3]Haddow A, Watkinson JM, Paterson E, Koller PC. Influence of Synthetic Oestrogens on Advanced Malignant Disease. British medical journal 1944; 2(4368): 393– 8. 20785660 PMC2286289
  4. [4]Ingle JN, Ahmann DL, Green SJ, Edmonson JH, Bisel HF, Kvols LK, Randomized clinical trial of diethylstilbestrol versus tamoxifen in postmenopausal women with advanced breast cancer. The New England journal of medicine 1981; 304(1): 16– 21. 7001242
  5. [5]Peethambaram PP, Ingle JN, Suman VJ, Hartmann LC, Loprinzi CL. Randomized trial of diethylstilbestrol vs. tamoxifen in postmenopausal women with metastatic breast cancer. An updated analysis. Breast Cancer Res Treat 1999; 54(2): 117– 22. 10424402
  6. [6]Kennedy BJ. Systemic Effects of Androgenic and Estrogenic Hormones in Advanced Breast Cancer. J Am Geriatr Soc 1965; 13: 230– 5. 14270629
  7. [7]Ellis MJ, Gao F, Dehdashti F, Jeffe DB, Marcom PK, Carey LA, Lower-dose vs high-dose oral estradiol therapy of hormone receptor-positive, aromatase inhibitor-resistant advanced breast cancer: a phase 2 randomized study. Jama 2009; 302(7): 774– 80. 19690310 PMC3460383
  8. [8]Iwase H, Yamamoto Y, Yamamoto-Ibusuki M, Murakami KI, Okumura Y, Tomita S, Ethinylestradiol is beneficial for postmenopausal patients with heavily pre-treated metastatic breast cancer after prior aromatase inhibitor treatment: a prospective study. British journal of cancer 2013; 109(6): 1537– 42. 24002591 PMC3777011
  9. [9]Zucchini G, Armstrong AC, Wardley AM, Wilson G, Misra V, Seif M, A phase II trial of low-dose estradiol in postmenopausal women with advanced breast cancer and acquired resistance to aromatase inhibition. European journal of cancer 2015; 51(18): 2725– 31. 26597446
  10. [10]Lonning PE, Taylor PD, Anker G, Iddon J, Wie L, Jorgensen LM, High-dose estrogen treatment in postmenopausal breast cancer patients heavily exposed to endocrine therapy. Breast Cancer Res Treat 2001; 67(2): 111– 6. 11519859
  11. [11]Agrawal A, Robertson JF, Cheung KL. Efficacy and tolerability of high dose “ethinylestradiol” in post-menopausal advanced breast cancer patients heavily pre-treated with endocrine agents. World journal of surgical oncology 2006; 4: 44. 16834778 PMC1533829
  12. [12]Traphagen NA, Hosford SR, Jiang A, Marotti JD, Brauer BL, Demidenko E, High estrogen receptor alpha activation confers resistance to estrogen deprivation and is required for therapeutic response to estrogen in breast cancer. Oncogene 2021; 40(19): 3408– 21. 33875787 PMC8122072
  13. [13]Miller TW, Hennessy BT, Gonzalez-Angulo AM, Fox EM, Mills GB, Chen H, Hyperactivation of phosphatidylinositol-3 kinase promotes escape from hormone dependence in estrogen receptor-positive human breast cancer. J Clin Invest 2010; 120(7): 2406– 13. 20530877 PMC2898598
  14. [14]Schwartz GN, Kaufman PA, Giridhar KV, Marotti JD, Chamberlin MD, Arrick BA, Alternating 17beta-estradiol and aromatase inhibitor therapies is efficacious in post-menopausal women with advanced endocrine-resistant ER+ breast cancer. Clin Cancer Res 2023. PMC10688025 37260292
  15. [15]Li S, Shen D, Shao J, Crowder R, Liu W, Prat A, Endocrine-therapy-resistant ESR1 variants revealed by genomic characterization of breast-cancer-derived xenografts. Cell reports 2013; 4(6): 1116– 30. 24055055 PMC3881975
  16. [16]DeRose YS, Gligorich KM, Wang G, Georgelas A, Bowman P, Courdy SJ, Patient-derived models of human breast cancer: protocols for in vitro and in vivo applications in tumor biology and translational medicine. Curr Protoc Pharmacol 2013; Chapter 14:Unit14 23. PMC3630511 23456611
  17. [17]Martin M Cutadapt removes adapter sequences from high-throughput sequencing reads. 2011 2011; 17(1): 3.
  18. [18]Li W, Koster J, Xu H, Chen CH, Xiao T, Liu JS, Quality control, modeling, and visualization of CRISPR screens with MAGeCK-VISPR. Genome biology 2015; 16: 281. 26673418 PMC4699372
  19. [20]Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, STAR: ultrafast universal RNA-seq aligner. Bioinformatics 2013; 29(1): 15– 21. 23104886 PMC3530905
  20. [21]Barbie DA, Tamayo P, Boehm JS, Kim SY, Moody SE, Dunn IF, Systematic RNA interference reveals that oncogenic KRAS-driven cancers require TBK1. Nature 2009; 462(7269): 108– 12. 19847166 PMC2783335
  21. [22]Reich M, Liefeld T, Gould J, Lerner J, Tamayo P, Mesirov JP. GenePattern 2.0. Nature genetics 2006; 38(5): 500– 1. 16642009
  22. [23]Demidenko E Mixed models: theory and applications with R. Hoboken, New Jersey: Wiley; 2013. xxvii, 717 pages p.
  23. [24]Demidenko E Advanced statistics with applications in R. Wiley series in probability and statistics. Hoboken, NJ: Wiley,; 2020. p 1 online resource.
  24. [25]Demidenko E, Miller TW. Statistical determination of synergy based on Bliss definition of drugs independence. Plos One 2019; 14(11): e0224137. 31765385 PMC6876842
  25. [26]Demidenko E Mixed Models: Theory and Applications with R. Hoboken, NJ: Wiley; 2013.
  26. [27]Demidenko E Advanced Statistics with Applications in R. Wiley (Hoboken, NJ). 2020.
  27. [28]Hosford SR, Shee K, Wells JD, Traphagen NA, Fields JL, Hampsch RA, Estrogen therapy induces an unfolded protein response to drive cell death in ER+ breast cancer. Molecular oncology 2019; 13(8): 1778– 94. 31180176 PMC6670014
  28. [29]Pakdel F, Le Goff P, Katzenellenbogen BS. An assessment of the role of domain F and PEST sequences in estrogen receptor half-life and bioactivity. The Journal of steroid biochemistry and molecular biology 1993; 46(6): 663– 72. 8274400
  29. [30]Kumarasamy V, Vail P, Nambiar R, Witkiewicz AK, Knudsen ES. Functional Determinants of Cell Cycle Plasticity and Sensitivity to CDK4/6 Inhibition. Cancer Res 2021; 81(5): 1347– 60. 33323381 PMC8026500
  30. [31]Mailand N, Gibbs-Seymour I, Bekker-Jensen S. Regulation of PCNA-protein interactions for genome stability. Nature reviews Molecular cell biology 2013; 14(5): 269– 82. 23594953
  31. [32]Ginno PA, Lott PL, Christensen HC, Korf I, Chedin F. R-loop formation is a distinctive characteristic of unmethylated human CpG island promoters. Mol Cell 2012; 45(6): 814– 25. 22387027 PMC3319272
  32. [33]Skourti-Stathaki K, Proudfoot NJ, Gromak N. Human senataxin resolves RNA/DNA hybrids formed at transcriptional pause sites to promote Xrn2-dependent termination. Mol Cell 2011; 42(6): 794– 805. 21700224 PMC3145960
  33. [34]Yang Y, McBride KM, Hensley S, Lu Y, Chedin F, Bedford MT. Arginine methylation facilitates the recruitment of TOP3B to chromatin to prevent R loop accumulation. Mol Cell 2014; 53(3): 484– 97. 24507716 PMC3959860
  34. [35]Hamperl S, Bocek MJ, Saldivar JC, Swigut T, Cimprich KA. Transcription-Replication Conflict Orientation Modulates R-Loop Levels and Activates Distinct DNA Damage Responses. Cell 2017; 170(4): 774– 86 e19. 28802045 PMC5570545
  35. [36]Stork CT, Bocek M, Crossley MP, Sollier J, Sanz LA, Chedin F, Co-transcriptional R-loops are the main cause of estrogen-induced DNA damage. eLife 2016; 5. PMC5030092 27552054
  36. [37]Ye BJ, Kang HJ, Lee-Kwon W, Kwon HM, Choi SY. PARP1-mediated PARylation of TonEBP prevents R-loop-associated DNA damage. DNA Repair (Amst) 2021; 104: 103132. 34049076
  37. [38]Cristini A, Groh M, Kristiansen MS, Gromak N. RNA/DNA Hybrid Interactome Identifies DXH9 as a Molecular Player in Transcriptional Termination and R-Loop-Associated DNA Damage. Cell reports 2018; 23(6): 1891– 905. 29742442 PMC5976580
  38. [40]Moynahan ME, Pierce AJ, Jasin M. BRCA2 is required for homology-directed repair of chromosomal breaks. Mol Cell 2001; 7(2): 263– 72. 11239455
  39. [41]Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA, Richardson TB, Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature 2005; 434(7035): 917– 21. 15829967
  40. [42]Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature 2005; 434(7035): 913– 7. 15829966
  41. [43]Kota K, Brufsky A, Oesterreich S, Lee A. Estradiol as a Targeted, Late-Line Therapy in Metastatic Breast Cancer with Estrogen Receptor Amplification. Cureus 2017; 9(7): e1434. 28924522 PMC5587405
  42. [44]Savage KI, Matchett KB, Barros EM, Cooper KM, Irwin GW, Gorski JJ, BRCA1 deficiency exacerbates estrogen-induced DNA damage and genomic instability. Cancer Res 2014; 74(10): 2773– 84. 24638981 PMC4024319
  43. [46]Tripathi K, Mani C, Somasagara RR, Clark DW, Ananthapur V, Vinaya K, Detection and evaluation of estrogen DNA-adducts and their carcinogenic effects in cultured human cells using biotinylated estradiol. Mol Carcinog 2017; 56(3): 1010– 20. 27597267
  44. [47]Ju BG, Lunyak VV, Perissi V, Garcia-Bassets I, Rose DW, Glass CK, A topoisomerase IIbeta-mediated dsDNA break required for regulated transcription. Science 2006; 312(5781): 1798– 802. 16794079
  45. [48]Bhardwaj P, Iyengar NM, Zahid H, Carter KM, Byun DJ, Choi MH, Obesity promotes breast epithelium DNA damage in women carrying a germline mutation in BRCA1 or BRCA2. Science translational medicine 2023; 15(684):eade1857. PMC10557057 36812344
  46. [49]Chatterjee P, Schweizer MT, Lucas JM, Coleman I, Nyquist MD, Frank SB, Supraphysiological androgens suppress prostate cancer growth through androgen receptor-mediated DNA damage. J Clin Invest 2019; 130: 4245– 60. PMC6763228 31310591
  47. [50]Schweizer MT, Antonarakis ES, Wang H, Ajiboye AS, Spitz A, Cao H, Effect of bipolar androgen therapy for asymptomatic men with castration-resistant prostate cancer: results from a pilot clinical study. Sci Transl Med 2015; 7(269): 269ra2. PMC4507510 25568070
  48. [51]Schweizer MT, Gulati R, Yezefski T, Cheng HH, Mostaghel E, Haffner MC, Bipolar androgen therapy plus olaparib in men with metastatic castration-resistant prostate cancer. Prostate Cancer Prostatic Dis 2022. PMC10286318 36564459
  49. [52]Rebbeck TR, Mitra N, Wan F, Sinilnikova OM, Healey S, McGuffog L, Association of type and location of BRCA1 and BRCA2 mutations with risk of breast and ovarian cancer. JAMA 2015; 313(13): 1347– 61. 25849179 PMC4537700
  50. [53]Dean JL, McClendon AK, Knudsen ES. Modification of the DNA damage response by therapeutic CDK4/6 inhibition. J Biol Chem 2012; 287(34): 29075– 87. 22733811 PMC3436568
  51. [54]Pikman Y, Alexe G, Roti G, Conway AS, Furman A, Lee ES, Synergistic Drug Combinations with a CDK4/6 Inhibitor in T-cell Acute Lymphoblastic Leukemia. Clin Cancer Res 2017; 23(4): 1012– 24. 28151717 PMC5432118
  52. [55]Cao J, Zhu Z, Wang H, Nichols TC, Lui GYL, Deng S, Combining CDK4/6 inhibition with taxanes enhances anti-tumor efficacy by sustained impairment of pRB-E2F pathways in squamous cell lung cancer. Oncogene 2019; 38(21): 4125– 41. 30700828
  53. [56]Salvador-Barbero B, Alvarez-Fernandez M, Zapatero-Solana E, El Bakkali A, Menendez MDC, Lopez-Casas PP, CDK4/6 Inhibitors Impair Recovery from Cytotoxic Chemotherapy in Pancreatic Adenocarcinoma. Cancer Cell 2020; 38(4): 584. 33049208

Cited by

Sorted by citations

Authors
Traphagen, N. A., Schwartz, G. N., Tau, S., Roberts, A. M., Jiang, A., Hosford, S. R., Marotti, J. D., Goen, A. E., Romo, B. A., Johnson, A. L., Duffy, E.-C. K., Demidenko, E., Heverly, P., Mosesson, Y., Soucy, S. M., Kolling, F., & Miller, T. W.
Affiliations
  • Dartmouth College (US)
  • 5Champions Oncology Inc., Rockville, Maryland.
Corresponding author
Miller, Todd W.
ORCID
Venue
Clinical Cancer Research
Publisher
American Association for Cancer Research (AACR)
Published
2023-07-26
Volume
29
Issue
18
Pages
3717-3728
ISSN
  • 1078-0432 (ISSN-L)
  • 1557-3265
DOI
10.1158/1078-0432.ccr-23-0488
PMC
PMC10528687
OpenAlex
W4384126490
Cited by
20
References
3
Citations by year
2026: 9 · 2025: 4 · 2024: 3 · 2023: 4
FWCI
2.44
Type
journal article
Language
en
Open access
Locations
  • Clinical Cancer Research (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • PubMed Central (submittedVersion)
Funders
  • National Cancer Institute (R01CA200994, R01CA267691, R01CA262232, R01CA211869, P30CA023108, F31CA243409, S10OD030242)
  • Susan G. Komen (CCR1533084)
  • National Institute of General Medical Sciences (P20GM130454)
Topics
  • Health Sciences › Medicine › Oncology › PARP inhibition in cancer therapy
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
  • Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Advanced Breast Cancer Therapies
Keywords
estrogen therapy, estrogen receptor alpha, estrogen receptor-positive breast cancer, endocrine therapy resistance, estradiol, DNA damage response, PARP inhibition, olaparib, R-loops, RNA-DNA hybrids, transcriptional stress, BRCA mutations, combination therapy, CRISPR-Cas9 screen

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