Breast cancer is the most prevalent cancer in women, and over two-thirds of cases express estrogen receptor-α (ER-α, encoded by ESR1). Through a prospective clinical sequencing program for advanced cancers, we enrolled 11 patients with ER-positive metastatic breast cancer. Whole-exome and transcriptome analysis showed that six cases harbored mutations of ESR1 affecting its ligand-binding domain (LBD), all of whom had been treated with anti-estrogens and estrogen deprivation therapies. A survey of The Cancer Genome Atlas (TCGA) identified four endometrial cancers with similar mutations of ESR1. The five new LBD-localized ESR1 mutations identified here (encoding p.Leu536Gln, p.Tyr537Ser, p.Tyr537Cys, p.Tyr537Asn and p.Asp538Gly) were shown to result in constitutive activity and continued responsiveness to anti-estrogen therapies in vitro. Taken together, these studies suggest that activating mutations in ESR1 are a key mechanism in acquired endocrine resistance in breast cancer therapy.

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The authors thank D. Miller, T. Barrette and D. Gibbs for hardware and database management, K. Giles for assistance with manuscript preparation, physicians M. Wicha, L. Pierce, D. Smith, K. Levin and F. Feng for referring patients, and C. Betts and J. Athanikar for assistance with tumor boards. We also thank the larger MI-ONCOSEQ team, including clinical research coordinator E. Williams, pathologist R. Mehra, genetic counselors J. Everett, S. Gustafson and V. Raymond, and radiologists E. Higgins, E. Caoili and R. Dunnick. This project is supported in part by the Prostate Cancer Foundation with funding for our sequencing infrastructure, the National Cancer Institute Early Detection Research Network (U01 CA111275), National Human Genome Research Institute Clinical Sequencing Exploratory Research (CSER) Consortium (1UM1HG006508), US Department of Defense contract W81XWH-12-1-0080 and a Department of Defense Era of Hope Scholar Award. A.M.C. is also supported by the Alfred A. Taubman Institute, the American Cancer Society, the Howard Hughes Medical Institute and a Doris Duke Charitable Foundation Clinical Scientist Award.

Author information

Author notes

    • Dan R Robinson
    •  & Yi-Mi Wu

    These authors contributed equally to this work.


  1. Michigan Center for Translational Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Dan R Robinson
    • , Yi-Mi Wu
    • , Pankaj Vats
    • , Fengyun Su
    • , Robert J Lonigro
    • , Xuhong Cao
    • , Shanker Kalyana-Sundaram
    • , Rui Wang
    • , Yu Ning
    • , Lynda Hodges
    • , Amy Gursky
    • , Javed Siddiqui
    • , Scott A Tomlins
    • , Lakshmi P Kunju
    •  & Arul M Chinnaiyan
  2. Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Dan R Robinson
    • , Yi-Mi Wu
    • , Pankaj Vats
    • , Fengyun Su
    • , Shanker Kalyana-Sundaram
    • , Rui Wang
    • , Yu Ning
    • , Amy Gursky
    • , Javed Siddiqui
    • , Scott A Tomlins
    • , Lakshmi P Kunju
    •  & Arul M Chinnaiyan
  3. Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Robert J Lonigro
    • , James M Rae
    •  & Arul M Chinnaiyan
  4. Howard Hughes Medical Institute, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Xuhong Cao
    •  & Arul M Chinnaiyan
  5. Department of Internal Medicine, Ohio State University, Columbus, Ohio, USA.

    • Sameek Roychowdhury
  6. Brady Urological Institute, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

    • Kenneth J Pienta
  7. Center for Bioethics and Social Science in Medicine, University of Michigan, Ann Arbor, Michigan, USA.

    • Scott Y Kim
  8. Department of Health Behavior & Health Education, University of Michigan, Ann Arbor, Michigan, USA.

    • J Scott Roberts
  9. Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • James M Rae
    • , Catherine H Van Poznak
    • , Daniel F Hayes
    • , Rashmi Chugh
    • , Moshe Talpaz
    •  & Anne F Schott
  10. Department of Urology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Arul M Chinnaiyan
  11. Center for Computational Medicine and Biology, University of Michigan, Ann Arbor, Michigan, USA.

    • Arul M Chinnaiyan


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D.R.R., Y.-M.W. and A.M.C. conceived the experiments. D.R.R., Y.-M.W., X.C., R.W., F.S. and Y.N. performed exome and transcriptome sequencing. P.V., R.J.L., S.K.-S. and D.R.R. carried out bioinformatics analysis of high-throughput sequencing data for somatic mutation, copy number and tumor content determination and performed gene expression and gene fusion analysis. D.R.R., Y.-M.W. and F.S. generated ESR1 constructs and carried out in vitro experiments. L.H. coordinated patients for clinical research. J.S. and A.G. collected and processed clinical tissue samples for next-generation sequencing. L.P.K. and S.A.T. provided pathology review. J.M.R. provided experimental analysis. C.H.V.P., D.F.H., R.C. and A.F.S. enrolled patients and provided clinical data and consultation at tumor boards. D.R.R., X.C., Y.-M.W., P.V., R.J.L., S.K.-S., S.Y.K., J.S.R., S.R., M.T., K.J.P. and A.M.C. developed the integrated clinical sequencing protocol. D.R.R., Y.-M.W. and A.M.C. prepared the manuscript, which was reviewed by all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Arul M Chinnaiyan.

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