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Mutations in the seed region of human miR-96 are responsible for nonsyndromic progressive hearing loss

Abstract

MicroRNAs (miRNAs) bind to complementary sites in their target mRNAs to mediate post-transcriptional repression1,2, with the specificity of target recognition being crucially dependent on the miRNA seed region3. Impaired miRNA target binding resulting from SNPs within mRNA target sites has been shown to lead to pathologies associated with dysregulated gene expression4,5,6,7. However, no pathogenic mutations within the mature sequence of a miRNA have been reported so far. Here we show that point mutations in the seed region of miR-96, a miRNA expressed in hair cells of the inner ear8, result in autosomal dominant, progressive hearing loss. This is the first study implicating a miRNA in a mendelian disorder. The identified mutations have a strong impact on miR-96 biogenesis and result in a significant reduction of mRNA targeting. We propose that these mutations alter the regulatory role of miR-96 in maintaining gene expression profiles in hair cells required for their normal function.

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Figure 1: Mutations in the seed region of MIR96 cause DFNA50 hearing loss.
Figure 2: Predicted secondary structure and processing of the wild-type and mutant forms of miR96.
Figure 3: Downregulation of predicted primary targets is impaired by the miR-96 (+13G>A) and (+14C>A) mutations.

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Acknowledgements

We thank members of the Spanish families whose participation made this study possible. We also express our gratitude to A. Fatica and I. Bozzoni (Universita di Roma “La Sapienza”) for the donation of the microRNA expression vector and to P. Morales (Hospital 12 de Octubre) for providing control DNAs. S.M.-H. is recipient of a fellowship from L'Oréal-UNESCO “for women in Science”. A.M. is recipient of a contract from the Centre for Biomedical Research on Rare Diseases (CIBERER) and M.M. is recipient of a contract from the EuroHear project. F.M.-M. is a fellow from the Spanish Ministerio de Ciencia y Tecnología. This work was supported by grants from the Spanish Ministerio de Ciencia y Tecnología (SAF2005-06355 to F.M.), Spanish Fondo de Investigaciones anitarias (FIS CP03/00014 and PI08/0045 to M.A.M.-P; PI-051942; G03/203), the European Commission (FP6 Integrated Projects: SIROCCO LSHG-CT-2006-037900 to T.D.; EUROHEAR, LSHG-CT-2004-512063), Deafness Research UK and the Wellcome Trust.

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The exclusion of candidates genes within the DFNA50 genetic interval and the identification of the miR96 mutations were carried out by A.M., S.M.-H., M.M., F.M.-M. and L.O. Screening in autosomal recessive deafness families was designed and performed by I.d.C. and L.A.A. Vector generation for luciferase assays and miR96 expression was carried out by A.M., and the experiments were designed and performed by N.R. and T.D. Experimental design of the study and writing of the manuscript was carried out by M.A.M.-P. K.P.S. shared data and ideas. Scientific discussion of data and critical reading of the paper were performed by K.P.S., T.D., I.d.C., F.M. and M.A.M.-P.

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Correspondence to Miguel Ángel Moreno-Pelayo.

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Supplementary Figures 1–6, Supplementary Methods and Supplementary Tables 1–3 (PDF 415 kb)

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Mencía, Á., Modamio-Høybjør, S., Redshaw, N. et al. Mutations in the seed region of human miR-96 are responsible for nonsyndromic progressive hearing loss. Nat Genet 41, 609–613 (2009). https://doi.org/10.1038/ng.355

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