Abstract
RIPK3 is a key regulator of necroptosis, but the specific roles of its kinase-dependent and -independent functions in disease pathogenesis remain poorly understood. Here, we generated and characterized RIPK3 D143N kinase-dead knock-in mice, a novel kinase-inactive model that selectively disrupts RIPK3 kinase activity without inducing spontaneous apoptosis. Unlike previously reported kinase-inactive Ripk3D161N/D161N mice, which exhibit embryonic lethality by triggering apoptosis, Ripk3D143N/D143N mice are viable and fertile, demonstrating that RIPK3 kinase activity is dispensable for development. The RIPK3 D143N mutation effectively blocks necroptosis induced by multiple stimuli and fully rescues embryonic lethality of caspase-8-deficient mice. Notably, Ripk3D143N/D143N mice were significantly less protected from TNF-driven inflammatory disease than RIPK3-deficient mice, revealing a critical kinase-independent role for RIPK3. This scaffold function drives inflammation and tissue damage through JAK-STAT1 activation, as pharmacological inhibition of JAK1/2 effectively reduces disease pathogenesis. Thus, our findings establish Ripk3D143N/D143N mice as a valuable model for dissecting the kinase and scaffold functions of RIPK3, and highlights the therapeutic potential of targeting its scaffold function in inflammatory diseases.
Schematic diagram showing the kinase-dependent and kinase-independent function in cell death and inflammation. A novel kinase-inactive RIPK3 mouse model (Ripk3D143N/D143N) defines the kinase-dependent and kinase-independent role of RIPK3 in cell death and inflammation.
Data availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank Dr. Xiaodong Wang (National Institute of Biological Sciences (NIBS), Beijing, China) for kindly providing Smac mimetic and Ripk3-/- mice. This work was supported by the National Key Research and Development Program of China (No. 2022YFC2502700), the National Natural Science Foundation of China (31830051, 32370810, 82371876, 32500645), the CAMS Innovation Fund for Medical Sciences (2023-I2M-2-005, 2024-I2M-TS-032, 2025-I2M-XHJC-051, 2022-I2M-2-004, 2021-I2M-1-041, 2021-I2M-1-047, and 2021-I2M-1-061), Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences (2021-PT180-001, 2019PT310028, 2017NL31004, 2017NL31002), the Special Research Fund for Central Universities, Peking Union Medical College (3332022077, 3332025146), Basic Research Program of Jiangsu (BK20243030, BK20250445), the Suzhou Municipal Key Laboratory (SZS2022005), and the NCTIB Fund for R&D platform for Cell and Gene Therapy.
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S.D.H., X.Y. and X.Z. designed this study and wrote the manuscript. Y.D., J.L., C.Z. and S.Q.H. designed and performed the majority of the experiments, analyzed the data, and wrote the manuscript. Z.L. synthesized the chemical compound and analyzed the data, J.Q analyzed the bulk RNA-seq data, Q.L., X.X. and C.Z. provided technical assistance and expertise for TNF-induced SIRS experiments, L.X. and F.M. provided technical assistance for data analysis.
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X. Z. and S.D.H. are co-founders, consultants, and shareholders of Accro Bioscience Inc, which supports research in their labs. The remaining authors declare no competing financial interests.
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All animal experiments were performed in accordance with protocols approved by the Suzhou Institutes of Systems Medicine Institutional Animal Care and Use Committee (ISM-IACUC-0037-R).
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Du, Y., Li, J., Zhao, C. et al. A viable kinase-inactive RIPK3 D143N mouse model reveals its scaffold function in driving TNF-induced inflammatory disorder. Cell Death Discov. (2026). https://doi.org/10.1038/s41420-026-02962-x
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DOI: https://doi.org/10.1038/s41420-026-02962-x
