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Non-phosphorylatable cyclin D1 mutant potentiates endometrial hyperplasia and drives carcinoma with Pten loss

Oncogene volume 41, pages 2187–2195 (2022)Cite this article

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Abstract

Cyclin D1 is a regulatory subunit of -Cyclin Dependent Kinases 4 and 6 (CDK4/6) and regulates progression from G1 to S phase of the cell cycle. Dysregulated cyclin D1-CDK4/6 contributes to abnormal cell proliferation and tumor development. Phosphorylation of threonine 286 of cyclin D1 is necessary for ubiquitin-dependent degradation. Non-phosphorylatable cyclin D1 mutants are stabilized and concentrated in the nucleus, contributing to genomic instability and tumor development. Studies investigating the tumor-promoting functions of cyclin D1 mutants have focused on the use of artificial promoters to drive the expression which unfortunately may not accurately reflect tumorigenic functions of mutant cyclin D1 in cancer development. We have generated a conditional knock-in mouse model where cyclin D1T286A is expressed under the control of its endogenous promoter following Cre-dependent excision of a lox-stop-lox sequence. Acute expression of cyclin D1T286A following tamoxifen-inducible Cre recombinase triggers inflammation, lymphocyte abnormality and ultimately mesenteric tumors in the intestine. Tissue-specific expression of cyclin D1T286A in the uterus and endometrium cooperates with Pten loss to drive endometrial hyperplasia and cancer. Mechanistically, cyclin D1T286A mutant activates NF-κB signaling, augments inflammation, and contributes to tumor development. These results indicate that mutation of cyclin D1 at threonine 286 has a critical role in regulating inflammation and tumor development.

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Fig. 1: Characterization of cyclin D1 knock-in mouse model.
Fig. 2: Endogenous expression of cyclin D1T286A induces mesenteric tumors in mice.
Fig. 3: Cyclin D1T286A accelerates Pten + / − induced endometrial hyperplasia development and progression.
Fig. 4: NFkB signaling is activated upon cyclin D1 T286A expression.
Fig. 5: p65 (RelA) is regulated by cyclin D1-CDK4/6 complex.

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Acknowledgements

We thank assistance from Dr. E. Ricky Chan in the Institute for Computational Biology for RNA sequencing analysis, Simone Edelheit in the Genomics Core for RNA sequencing sample preparation, and the Animal Resource Center at Case Western Reserve University. We thank assistance from Melodie Parrish in the Translational Science Resource at The Medical University of South Carolina for Hematoxylin and Eosin staining and immunohistochemistry staining.

Funding

This work was supported by grants from National Cancer Institute: R01 CA093237 and P01 CA098101 (JAD); R01 CA121275 and Advancing a Healthier Wisconsin Endowment Award (GL).

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Authors and Affiliations

  1. Department of Dermatology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, OH, 44106, USA

    Akihiro Yoshida

  2. Department of Biochemistry, Case Western Reserve University, Cleveland, OH, 44106, USA

    Akihiro Yoshida, Polly Phillips-Mason, Vincenzo Tarallo & J. Alan Diehl

  3. Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, OH, 44106, USA

    Akihiro Yoshida, Stefanie Avril & J. Alan Diehl

  4. Department of Pathology, University Hospitals Cleveland Medical Center and Case Western Reserve University, Cleveland, OH, 44106, USA

    Stefanie Avril

  5. Department of Biochemistry, Medical University of South Carolina, Charleston, SC, 29425, USA

    Christopher Koivisto

  6. Medical College of Wisconsin Cancer Center, Department of Biochemistry, Medical College of Wisconsin, Wauwatosa, WI, 53226, USA

    Gustavo Leone

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  1. Akihiro Yoshida
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Contributions

AY and JAD designed experiments, analyzed data, and wrote the manuscript. AY, PP-M, and VT performed experiments. SA and CK performed histological analysis. GL provided Sprr2f-Cre transgenic mice.

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Correspondence to Akihiro Yoshida or J. Alan Diehl.

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Yoshida, A., Phillips-Mason, P., Tarallo, V. et al. Non-phosphorylatable cyclin D1 mutant potentiates endometrial hyperplasia and drives carcinoma with Pten loss. Oncogene 41, 2187–2195 (2022). https://doi.org/10.1038/s41388-022-02243-8

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