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SMAD4 mutations and cross-talk between TGF-β/IFNγ signaling accelerate rates of DNA damage and cellular senescence, resulting in a segmental progeroid syndrome—the Myhre syndrome

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Abstract

SMAD4 encodes a member of the SMAD family of proteins involved in the TGF-β signaling pathway. Potentially heritable, autosomal dominant, gain-of-function heterozygous variants of SMAD4 cause a rare developmental disorder, the Myhre syndrome, which is associated with a wide range of developmental and post-developmental phenotypes that we now characterize as a novel segmental progeroid syndrome. Whole-exome sequencing of a patient referred to our International Registry of Werner Syndrome revealed a heterozygous p.Arg496Cys variant of the SMAD4 gene. To investigate the role of SMAD4 mutations in accelerated senescence, we generated cellular models overexpressing either wild-type SMAD4 or mutant SMAD4-R496C in normal skin fibroblasts. We found that cells expressing the SMAD4-R496C mutant exhibited decreased proliferation and elevated expression of cellular senescence and inflammatory markers, including IL-6, IFNγ, and a TGF-β target gene, PAI-1. Here we show that transient exposure to TGF-β, an inflammatory cytokine, followed by chronic IFNγ stimulation, accelerated rates of senescence that were associated with increased DNA damage foci and SMAD4 expression. TGF-β, IFNγ, or combinations of both were not sufficient to reduce proliferation rates of fibroblasts. In contrast, TGF-β alone was able to induce preadipocyte senescence via induction of the mTOR protein. The mTOR inhibitor rapamycin mitigated TGF-β-induced expression of p21, p16, and DNA damage foci and improved replicative potential of preadipocytes, supporting the cell-specific response to this cytokine. These findings collectively suggest that persistent DNA damage and cross-talk between TGF-β/IFNγ pathways contribute to a series of molecular events leading to cellular senescence and a segmental progeroid syndrome.

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Data availability

Data that support the findings of this study are available from the corresponding author upon request.

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Acknowledgments

The authors are grateful to Ms. Lin Lee for administrative and laboratory assistance, Ms. Nino Giorgadze for isolation of primary human preadipocytes, and Dr. Tamar Pirtskhalava for insightful discussions.

Funding

This work was supported in part by JSPS KAKENHI 17H04037 (JO) and NIH grants R01CA210916 (GMM/JO), R37AG013925 (JLK/TT), and P01AG062413 (JLK/TT), and Robert and Arlene Kogod (JLK/TT), the Connor Group (JLK/TT), Robert J. and Theresa W. Ryan (JLK/TT), and the Noaber Foundation (JLK/TT).

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Authors and Affiliations

  1. Department of Pathology, University of Washington, Box 357470, HSB, Seattle, WA, K-543, USA

    Renuka Kandhaya-Pillai, Deyin Hou, Jiaming Zhang, Xiaomeng Yang, George M. Martin & Junko Oshima

  2. Department of Dermatology, University of Southern California, Los Angeles, CA, USA

    Goli Compoginis

  3. Department of Medicine, Tokyo Medical and Dental University, Tokyo, Japan

    Takayasu Mori

  4. Robert and Arlene Kogod Center on Aging, Mayo Clinic, Rochester, MN, USA

    Tamara Tchkonia & James L. Kirkland

  5. Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, WA, USA

    Fuki M. Hisama

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  1. Renuka Kandhaya-Pillai
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Contributions

RKP designed and performed experiments, analyzed data, and wrote the manuscript. DH, JZ, and YY performed experiments and analyzed the data. GP and FH contributed clinical description of the patients and interpretation of clinical data. TM conducted exome analysis. TT and JLK contributed preadipocyte studies, supervised the project, and prepared and revised the manuscript. GMM and JO supervised the project and designed the study and prepared and revised the manuscript.

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Correspondence to Junko Oshima.

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Kandhaya-Pillai, R., Hou, D., Zhang, J. et al. SMAD4 mutations and cross-talk between TGF-β/IFNγ signaling accelerate rates of DNA damage and cellular senescence, resulting in a segmental progeroid syndrome—the Myhre syndrome. GeroScience 43, 1481–1496 (2021). https://doi.org/10.1007/s11357-020-00318-6

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