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Inhibition of KLF7-Targeting MicroRNA 146b Promotes Sciatic Nerve Regeneration

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Abstract

A previous study has indicated that Krüppel-like factor 7 (KLF7), a transcription factor that stimulates Schwann cell (SC) proliferation and axonal regeneration after peripheral nerve injury, is a promising therapeutic transcription factor in nerve injury. We aimed to identify whether inhibition of microRNA-146b (miR-146b) affected SC proliferation, migration, and myelinated axon regeneration following sciatic nerve injury by regulating its direct target KLF7. SCs were transfected with miRNA lentivirus, miRNA inhibitor lentivirus, or KLF7 siRNA lentivirus in vitro. The expression of miR146b and KLF7, as well as SC proliferation and migration, were subsequently evaluated. In vivo, an acellular nerve allograft (ANA) followed by injection of GFP control vector or a lentiviral vector encoding an miR-146b inhibitor was used to assess the repair potential in a model of sciatic nerve gap. miR-146b directly targeted KLF7 by binding to the 3′-UTR, suppressing KLF7. Up-regulation of miR-146b and KLF7 knockdown significantly reduced the proliferation and migration of SCs, whereas silencing miR-146b resulted in increased proliferation and migration. KLF7 protein was localized in SCs in which miR-146b was expressed in vivo. Similarly, 4 weeks after the ANA, anti-miR-146b increased KLF7 and its target gene nerve growth factor cascade, promoting axonal outgrowth. Closer analysis revealed improved nerve conduction and sciatic function index score, and enhanced expression of neurofilaments, P0 (anti-peripheral myelin), and myelinated axon regeneration. Our findings provide new insight into the regulation of KLF7 by miR-146b during peripheral nerve regeneration and suggest a potential therapeutic strategy for peripheral nerve injury.

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Acknowledgements

We would like to thank the staff of the Experimental Animal Center of China Medical University for providing excellent animal care. This work was supported by the National Natural Science Foundation of China (81371362, 81641125, and 81500629), the Scientific Research Foundation of Heilongjiang Province, China (LC2017040), the Science Fund of Heilongjiang Provincial Health and Family Planning Commission, China (2016357 and 2016385), and the Basic Research Operating Expenses Program of Heilongjiang Provincial Universities, China (2017- KYYWFMY-0661).

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Authors and Affiliations

  1. Department of Anatomy, Mudanjiang College of Medicine, Mudanjiang, 157011, China

    Wen-Yuan Li, Yan-Cui Liu, Ping Sun, Xiao-Feng Zhu & Ying Wang

  2. The Affiliated Hongqi Hospital, Mudanjiang College of Medicine, Mudanjiang, 157011, China

    Wei-Ting Zhang & Hui-Ting Li

  3. Department of Pathology, Mudanjiang College of Medicine, Mudanjiang, 157011, China

    Yong-Xia Cheng

  4. Department of Pharmacy, Mudanjiang College of Medicine, Mudanjiang, 157011, China

    Feng-Guo Zhai

  5. Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, 46202, USA

    Ling-Xiao Deng

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  1. Wen-Yuan Li
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Li, WY., Zhang, WT., Cheng, YX. et al. Inhibition of KLF7-Targeting MicroRNA 146b Promotes Sciatic Nerve Regeneration. Neurosci. Bull. 34, 419–437 (2018). https://doi.org/10.1007/s12264-018-0206-x

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