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ResearchIn-Press PreviewCell biologyOncology Open Access | 10.1172/JCI199483

Targeting casein kinase 1α-mediated FADD phosphorylation restores sensitivity in select CDK4/6 inhibitor-resistant breast cancer models

Sahezeel Awadia,1 Elizabeth K. Ziemke,2 Nicole M. Curnutt,3 Emily Kirk,4 Julianne Thomas,4 Anna Zimmerman,4 Maya J. Mileski,4 Sundaresh Ram,5 Reine Abou Zeidane,6 Craig Galban,2 Christina M. Woo,3 Corey W. Speers,7 Judith Leopold,2 and Alnawaz Rehemtulla4

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Awadia, S. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Ziemke, E. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Curnutt, N. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Kirk, E. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Thomas, J. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Zimmerman, A. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Mileski, M. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Ram, S. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

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1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Galban, C. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Woo, C. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Speers, C. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Leopold, J. in: PubMed | Google Scholar

1Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, United States of America

2Department of Radiology, The University of Michigan Medical School, Ann Arbor, United States of America

3Department of Chemistry and Chemical Biology, Harvard University, Cambridge, United States of America

4Department of Radiation Oncology, The University of Michigan Medical School, Ann Arbor, United States of America

5Department of Radiology and Imaging Sciences, Emory University and Georgia Institute of Technology, Atlanta, United States of America

6Department of Radiation Oncology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, United States of America

7Department of Radiation Oncology, The University of Alabama at Birmingham, Birmingham, United States of America

Find articles by Rehemtulla, A. in: PubMed | Google Scholar |

Published July 28, 2026 - More info

J Clin Invest. https://doi.org/10.1172/JCI199483.
Copyright © 2026, Awadia et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Published July 28, 2026 - Version history
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Abstract

Resistance to CDK4/6 inhibitors (CDK4/6i) combined with endocrine therapy presents a major barrier to improving outcomes in ER+ breast cancer. We identified FADD phosphorylation at Ser194 (phospho-FADD) as a mediator of CDK4/6i resistance in the models examined. Phospho-FADD acted as a pseudosubstrate inhibitor of the APC/C-Cdh1 complex, promoting G1/S transition and bypassing the canonical CDK4/6-Rb-E2F pathway. This CDK4/6-independent pathway was associated with PI3K hyperactivation. Clinical relevance of this bypass pathway was supported by increased phospho-FADD and pAKT in 73% of paired patient biopsies at post-treatment recurrence, while the remaining cases exhibited high baseline phospho-FADD and pAKT with intrinsic non-response. Inhibition of FADD phosphorylation with the CK1α degrader DEG-77, or PI3K inhibition, restored CDK4/6i sensitivity in resistant cells. In CDK4/6i-refractory xenografts, CK1α degradation combined with CDK4/6i resulted in profound tumor regressions and increased progression-free survival compared with either single agent, including complete tumor regressions in 92% of tumors. These findings support CK1α-mediated FADD phosphorylation as a targetable resistance mechanism in a subset of CDK4/6i-resistant ER+/HER2– breast cancer.

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