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Znhit3 regulates p53/p21 signaling and governs cerebellar granule cell development

Cell Death & Differentiation (2026)Cite this article

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Abstract

Mutations in ZNHIT3 are strongly associated with progressive encephalopathy with edema, hypsarrhythmia and optic atrophy (PEHO syndrome), characterized by severe cerebellar atrophy and profound intellectual disability; however, their role in cerebellar development remains unknown. By developing spatiotemporally-regulated conditional Znhit3 knockout mice, we discovered that Znhit3 is essential for granule cell progenitor survival, proliferation, differentiation, and migration. Knockout of Znhit3 caused loss of granule cell progenitors due to apoptosis, premature cell-cycle exit, and migration arrest and resulted in progressive anterior-lobe atrophy and motor deficits. The granule cell progenitor-autonomous defects secondarily impaired Purkinje cell alignment, dendritic maturation, and synaptic organization. Transcriptomic analyses revealed activation of the p53/p21 pathway, rRNA processing defects, and nucleolar stress. Genetic or pharmacologic inhibition of p53/p21 signaling rescued granule cell progenitor development and restored cerebellar architecture in the Znhit3-knockout mice. Thus, ZNHIT3 is a critical regulator of ribosome biogenesis and cerebellar growth, suggesting nucleolar stress-p53/p21 signaling as a potential therapeutic target in ZNHIT3-related disorders.

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Fig. 1: Specific ablation of Znhit3 in GCPs leads to progressive cerebellar dysplasia.
Fig. 2: Znhit3 cKOM mice have abnormal motor behaviors.
Fig. 3: ZNHIT3 regulates survival, proliferation, differentiation, and migration of GCPs.
Fig. 4: Znhit3 deletion in GCPs disrupts Purkinje cell development, Bergmann glia alignment, and circuit integration.
Fig. 5: Znhit3 deficiency alters the expression of neuronal genes and apoptosis-related signaling pathways.
Fig. 6: ZNHIT3 modulates rRNA processing, nucleolar stress, and the p53/p21 pathway in vitro.
Fig. 7: Deletion of p53 rescues the phenotype in ZNHIT3-deficient mice.

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Data availability

All high-throughput data mentioned in the paper are publicly available from GEO under accession (GSE298423).

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Acknowledgements

The authors would like to thank Dr. Jin Zhang in Fudan University for valuable technical guidance and providing the M17 cell line used in this study. We thank all members of the Zhou laboratory for constructive discussion. W.Z was supported by National Natural Science Foundation of China (82130048). Q.R.L was supported by the CancerFree Kids Foundation.

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Authors and Affiliations

  1. Institute of Pediatrics, Children’s Hospital of Fudan University, Shanghai, China

    Fangbing Chen, Zhiruo Kang, Kaiyi Liu & Wenyi Yang

  2. Department of Pediatrics, University of Cincinnati School of Medicine, Cincinnati, OH, USA

    Q. Richard Lu

  3. Key Laboratory of Birth Defects, Children’s Hospital of Fudan University, Shanghai, China

    Wenhao Zhou & Yifeng Lin

  4. Division of Neonatology and Center for Newborn Care, Guangzhou Women and Children’s Medical Center, Guangzhou Medical University, Guangzhou, China

    Wenhao Zhou

  5. Institute of Pediatric Infection, Immunity, and Critical Care Medicine, Shanghai Children’s Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China

    Yifeng Lin

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Contributions

Conceptualization: W.Z., and Y.L.; Methodology: F.C., Z.K., and K.L.; Investigation: F.C., Z.K., and W.Y.; Writing-Original Draft: F.C. and Q.R.L.; Writing-Review & Editing: Q.R.L., W.Z., and Y.L.; Resources: W.Z. and Y.L.; Supervision: W.Z., and Y.L.; Funding Acquisition: W.Z.

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Correspondence to Wenhao Zhou or Yifeng Lin.

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The authors declare no competing interests.

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All methods were performed in accordance with the relevant guidelines and regulations. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of Children’s Hospital of Fudan University (No. 2023-215). All mice used in experiments were maintained in a pathogen-free vivarium with a 12-hour light/dark cycle with free access to normal chow food and water. Stringent measures were taken to mitigate animal suffering and reduce the overall number of animals utilized. This study does not directly involve human subjects or human data that requires ethical approval.

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Chen, F., Kang, Z., Liu, K. et al. Znhit3 regulates p53/p21 signaling and governs cerebellar granule cell development. Cell Death Differ (2026). https://doi.org/10.1038/s41418-026-01707-8

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