Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss
Age-related hearing loss (ARHL) is the most common sensory impairment in the elderly affecting millions of people worldwide. To shed light on the genetics of ARHL, a large cohort of 464 Italian patients has been deeply characterized at clinical and molecular level. In particular, 46 candidate genes,...
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doaj-1bb308bee0c545e8a52ee95807cae31c2020-11-24T21:42:58ZengFrontiers Media S.A.Frontiers in Genetics1664-80212019-02-011010.3389/fgene.2019.00142420326Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing LossGiorgia Girotto0Giorgia Girotto1Anna Morgan2Anna Morgan3Navaneethakrishnan Krishnamoorthy4Navaneethakrishnan Krishnamoorthy5Massimiliano Cocca6Marco Brumat7Marco Brumat8Sissy Bassani9Sissy Bassani10Martina La Bianca11Mariateresa Di Stazio12Mariateresa Di Stazio13Paolo Gasparini14Paolo Gasparini15Department of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyDepartment of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalySidra Medical and Research Center, Doha, QatarHeart Science Centre, National Heart and Lung Institute, Imperial College London, London, United KingdomInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyDepartment of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyDepartment of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyDepartment of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyDepartment of Medicine, Surgery and Health Sciences, University of Trieste, Trieste, ItalyInstitute for Maternal and Child Health – IRCCS “Burlo Garofolo”, Trieste, ItalyAge-related hearing loss (ARHL) is the most common sensory impairment in the elderly affecting millions of people worldwide. To shed light on the genetics of ARHL, a large cohort of 464 Italian patients has been deeply characterized at clinical and molecular level. In particular, 46 candidate genes, selected on the basis of genome-wide association studies (GWAS), animal models and literature updates, were analyzed by targeted re-sequencing. After filtering and prioritization steps, SLC9A3R1 has been identified as a strong candidate and then validated by “in vitro” and “in vivo” studies. Briefly, a rare (MAF: 2.886e-5) missense variant c.539G > A, p.(R180Q) was detected in two unrelated male patients affected by ARHL characterized by a severe to profound high-frequency hearing loss. The variant, predicted as damaging, was not present in healthy matched controls. Protein modeling confirmed the pathogenic effect of p.(R180Q) variant on protein’s structure leading to a change in the total number of hydrogen bonds. In situ hybridization showed slc9a3r1 expression in zebrafish inner ear. A zebrafish knock-in model, generated by CRISPR-Cas9 technology, revealed a reduced auditory response at all frequencies in slc9a3r1R180Q/R180Q mutants compared to slc9a3r1+/+ and slc9a3r1+/R180Q animals. Moreover, a significant reduction (5.8%) in the total volume of the saccular otolith (which is responsible for sound detection) was observed in slc9a3r1R180Q/R180Q compared to slc9a3r1+/+ (P = 0.0014), while the utricular otolith, necessary for balance, was not affected in agreement with the human phenotype. Overall, these data strongly support the role of SLC9A3R1 gene in the pathogenesis of ARHL opening new perspectives in terms of diagnosis, prevention and treatment.https://www.frontiersin.org/article/10.3389/fgene.2019.00142/fullhearing lossnew gene discoveryzebrafish modelCRISPR-Cas9next-generation sequencing |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Giorgia Girotto Giorgia Girotto Anna Morgan Anna Morgan Navaneethakrishnan Krishnamoorthy Navaneethakrishnan Krishnamoorthy Massimiliano Cocca Marco Brumat Marco Brumat Sissy Bassani Sissy Bassani Martina La Bianca Mariateresa Di Stazio Mariateresa Di Stazio Paolo Gasparini Paolo Gasparini |
spellingShingle |
Giorgia Girotto Giorgia Girotto Anna Morgan Anna Morgan Navaneethakrishnan Krishnamoorthy Navaneethakrishnan Krishnamoorthy Massimiliano Cocca Marco Brumat Marco Brumat Sissy Bassani Sissy Bassani Martina La Bianca Mariateresa Di Stazio Mariateresa Di Stazio Paolo Gasparini Paolo Gasparini Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss Frontiers in Genetics hearing loss new gene discovery zebrafish model CRISPR-Cas9 next-generation sequencing |
author_facet |
Giorgia Girotto Giorgia Girotto Anna Morgan Anna Morgan Navaneethakrishnan Krishnamoorthy Navaneethakrishnan Krishnamoorthy Massimiliano Cocca Marco Brumat Marco Brumat Sissy Bassani Sissy Bassani Martina La Bianca Mariateresa Di Stazio Mariateresa Di Stazio Paolo Gasparini Paolo Gasparini |
author_sort |
Giorgia Girotto |
title |
Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss |
title_short |
Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss |
title_full |
Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss |
title_fullStr |
Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss |
title_full_unstemmed |
Next Generation Sequencing and Animal Models Reveal SLC9A3R1 as a New Gene Involved in Human Age-Related Hearing Loss |
title_sort |
next generation sequencing and animal models reveal slc9a3r1 as a new gene involved in human age-related hearing loss |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Genetics |
issn |
1664-8021 |
publishDate |
2019-02-01 |
description |
Age-related hearing loss (ARHL) is the most common sensory impairment in the elderly affecting millions of people worldwide. To shed light on the genetics of ARHL, a large cohort of 464 Italian patients has been deeply characterized at clinical and molecular level. In particular, 46 candidate genes, selected on the basis of genome-wide association studies (GWAS), animal models and literature updates, were analyzed by targeted re-sequencing. After filtering and prioritization steps, SLC9A3R1 has been identified as a strong candidate and then validated by “in vitro” and “in vivo” studies. Briefly, a rare (MAF: 2.886e-5) missense variant c.539G > A, p.(R180Q) was detected in two unrelated male patients affected by ARHL characterized by a severe to profound high-frequency hearing loss. The variant, predicted as damaging, was not present in healthy matched controls. Protein modeling confirmed the pathogenic effect of p.(R180Q) variant on protein’s structure leading to a change in the total number of hydrogen bonds. In situ hybridization showed slc9a3r1 expression in zebrafish inner ear. A zebrafish knock-in model, generated by CRISPR-Cas9 technology, revealed a reduced auditory response at all frequencies in slc9a3r1R180Q/R180Q mutants compared to slc9a3r1+/+ and slc9a3r1+/R180Q animals. Moreover, a significant reduction (5.8%) in the total volume of the saccular otolith (which is responsible for sound detection) was observed in slc9a3r1R180Q/R180Q compared to slc9a3r1+/+ (P = 0.0014), while the utricular otolith, necessary for balance, was not affected in agreement with the human phenotype. Overall, these data strongly support the role of SLC9A3R1 gene in the pathogenesis of ARHL opening new perspectives in terms of diagnosis, prevention and treatment. |
topic |
hearing loss new gene discovery zebrafish model CRISPR-Cas9 next-generation sequencing |
url |
https://www.frontiersin.org/article/10.3389/fgene.2019.00142/full |
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