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Bmi1 Regulates the Proliferation of Cochlear Supporting Cells Via the Canonical Wnt Signaling Pathway

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Abstract

Cochlear supporting cells (SCs), which include the cochlear progenitor cells, have been shown to be a promising resource for hair cell (HC) regeneration, but the mechanisms underlying the initiation and regulation of postnatal cochlear SC proliferation are not yet fully understood. Bmi1 is a member of the Polycomb protein family and has been reported to regulate the proliferation of stem cells and progenitor cells in multiple organs. In this study, we investigated the role of Bmi1 in regulating SC and progenitor cell proliferation in neonatal mice cochleae. We first showed that knockout of Bmi1 significantly inhibited the proliferation of SCs and Lgr5-positive progenitor cells after neomycin injury in neonatal mice in vitro, and we then showed that Bmi1 deficiency significantly reduced the sphere-forming ability of the organ of Corti and Lgr5-positive progenitor cells in neonatal mice. These results suggested that Bmi1 is required for the initiation of SC and progenitor cell proliferation in neonatal mice. Next, we found that DKK1 expression was significantly upregulated, while beta-catenin and Lgr5 expression were significantly downregulated in neonatal Bmi1−/− mice compared to wild-type controls. The observation that Bmi1 knockout downregulates Wnt signaling provides compelling evidence that Bmi1 is required for the Wnt signaling pathway. Furthermore, the exogenous Wnt agonist BIO overcame the downregulation of SC proliferation in Bmi1−/− mice, suggesting that Bmi1 knockout might inhibit the proliferation of SCs via downregulation of the canonical Wnt signaling pathway. Our findings demonstrate that Bmi1 plays an important role in regulating the proliferation of cochlear SCs and Lgr5-positive progenitor cells in neonatal mice through the Wnt signaling pathway, and this suggests that Bmi1 might be a new therapeutic target for HC regeneration.

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Acknowledgments

This work was supported by grants from the Major State Basic Research Development Program of China (973 Program) (2011CB504500, 2015CB965000), the National Natural Science Foundation of China (Nos. 81570913, 81470692, 81371094, 81230019, 81500790, 81570921, 31500852, 31501194), the Jiangsu Province Natural Science Foundation (BK20150022, BK20140620, BK20150598), the Project from China Scholarship Council (201406105052), the Program of Leading Medical Personnel in Shanghai, the Fundamental Research Funds for the Central Universities (2242014R30022, 021414380037), the Construction Program of Shanghai Committee of Science and Technology (12DZ2251700), and the Major Program of Shanghai Committee of Science and Technology (11441901000).

Author Contributions

HL and RC conceived and designed the experiments. XL, SS, QJ, ZS, WL, LL, and YZ performed the experiments. XL, SS, RC, and HL analyzed the data. DM provided the Bmi1 KO mice. XL, RC, and HL wrote the paper.

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    Authors and Affiliations

    1. Department of Otorhinolaryngology and Hearing Research Institute of Affiliated Eye and ENT Hospital, State Key Laboratory of Medicine Neurobiology, Fudan University, Room 611, Building 9, No. 83, Fenyang Road, Xuhui District, Shanghai, 200031, China

      Xiaoling Lu, Shan Sun, Wenyan Li, Liman Liu, Yanping Zhang, Yan Chen & Huawei Li

    2. Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Southeast University, Nanjing, 210096, China

      Jieyu Qi, Shasha Zhang & Renjie Chai

    3. Co-innovation Center of Neuroregeneration, Nantong University, Nantong, 226001, China

      Jieyu Qi, Shasha Zhang & Renjie Chai

    4. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China

      Lei Wang & Huawei Li

    5. State Key Laboratory of Reproductive Medicine, Research Center for Bone and Stem Cells, Department of Human Anatomy, Nanjing Medical University, Nanjing, 210096, China

      Dengshun Miao

    Authors
    1. Xiaoling Lu
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    2. Shan Sun
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    4. Wenyan Li
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    Corresponding authors

    Correspondence to Renjie Chai or Huawei Li.

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    The authors declare that they have no competing interests.

    Additional information

    Xiaoling Lu and Shan Sun contributed equally to this work.

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    Supp Fig 1

    Bmi1 is expressed in both HCs and SCs in the apical turn of the organ of Corti. Scale bars = 20 μm (DOCX 870 kb)

    Supp Fig 2

    Without damage, Lgr5 was expressed in Deiters’ cells, pillar cells, and inner phalangeal/border cells in neonatal WT mice. Scale bars = 20 μm (DOCX 717 kb)

    Supp Fig 3

    a Compared with P0 WT mice, the expression level of beta-catenin showed no difference in P0 Bmi1+/− mice. b The expression level of p27Kip1 in heterozygous mice showed no difference compared to P0 control. Scale bars = 20 μm (DOCX 809 kb)

    Supp Fig 4

    a The expression level of Lgr5 between P0 WT and Bmi1+/− mice showed no difference. b The expression level of Lgr5 between P30 WT and Bmi1+/− mice showed no difference. Scale bars = 20 μm. (DOCX 836 kb)

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    Lu, X., Sun, S., Qi, J. et al. Bmi1 Regulates the Proliferation of Cochlear Supporting Cells Via the Canonical Wnt Signaling Pathway. Mol Neurobiol 54, 1326–1339 (2017). https://doi.org/10.1007/s12035-016-9686-8

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    • DOI: https://doi.org/10.1007/s12035-016-9686-8

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