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Gene Ther. 2019 Dec;26(12):455-464. doi: 10.1038/s41434-019-0098-z. Epub 2019 Aug 27.

Downregulation of the human peripheral myelin protein 22 gene by miR-29a in cellular models of Charcot-Marie-Tooth disease.

Author information

1
Department of Molecular Genetics & Microbiology, College of Medicine University of Florida, Gainesville, FL, 32610, USA.
2
Department of Neuroscience, College of Medicine University of Florida, Gainesville, FL, 32610, USA.
3
Center for Translational Research in Neurodegenerative Disease, College of Medicine University of Florida, Gainesville, FL, 32610, USA.
4
UF Health Cancer Center, College of Medicine University of Florida, Gainesville, FL, 32610, USA.
5
Department of Neuroscience, College of Medicine University of Florida, Gainesville, FL, 32610, USA. notterp@mbi.ufl.edu.
6
Center for Translational Research in Neurodegenerative Disease, College of Medicine University of Florida, Gainesville, FL, 32610, USA. notterp@mbi.ufl.edu.

Abstract

The majority of hereditary neuropathies are caused by duplication of the peripheral myelin protein 22 (PMP22) gene. Therefore, mechanisms to suppress the expression of the PMP22 gene have high therapeutic significance. Here we asked whether the human PMP22 gene is a target for regulation by microRNA 29a (miR-29a). Using bioinformatics, we determined that the human PMP22 gene contains the conserved seed sequence of the miR-29a binding site and this regulatory motif is included in the duplicated region in neuropathic patients. Using luciferase reporter assays in HEK293 cells, we demonstrated that transient transfection of a miR-29a mimic is associated with reduction in PMP22 3'UTR reporter activity. Transfecting normal and humanized transgenic neuropathic mouse Schwann cells with a miR-29a expression plasmid effectively lowered both the endogenous mouse and the transgenic human PMP22 transcripts compared with control vector. In dermal fibroblasts derived from neuropathic patients, ectopic expression of miR-29a led to ~50% reduction in PMP22 mRNA, which corresponded to ~20% decrease in PMP22 protein levels. Significantly, miR-29a-mediated reduction in PMP22 mitigated the reduced mitotic capacity of the neuropathic cells. Together, these results support further testing of miR-29a and/or PMP22-targeting siRNAs as therapeutic agents for correcting the aberrant expression of PMP22 in neuropathic patients.

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