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Human Genetics

, Volume 135, Issue 5, pp 513–524 | Cite as

A mutation in SLC22A4 encoding an organic cation transporter expressed in the cochlea strial endothelium causes human recessive non-syndromic hearing loss DFNB60

  • Mariem Ben Said
  • M’hamed Grati
  • Takahiro Ishimoto
  • Bing Zou
  • Imen Chakchouk
  • Qi Ma
  • Qi Yao
  • Bouthaina Hammami
  • Denise Yan
  • Rahul Mittal
  • Noritaka Nakamichi
  • Abdelmonem Ghorbel
  • Lingling Neng
  • Mustafa Tekin
  • Xiao Rui Shi
  • Yukio Kato
  • Saber MasmoudiEmail author
  • Zhongmin Lu
  • Mounira Hmani
  • Xuezhong LiuEmail author
Original Investigation

Abstract

The high prevalence/incidence of hearing loss (HL) in humans makes it the most common sensory defect. The majority of the cases are of genetic origin. Non-syndromic hereditary HL is extremely heterogeneous. Genetic approaches have been instrumental in deciphering genes that are crucial for auditory function. In this study, we first used NADf chip to exclude the implication of known North-African mutations in HL in a large consanguineous Tunisian family (FT13) affected by autosomal recessive non-syndromic HL (ARNSHL). We then performed genome-wide linkage analysis and assigned the deafness gene locus to ch:5q23.2-31.1, corresponding to the DFNB60 ARNSHL locus. Moreover, we performed whole exome sequencing on FT13 patient DNA and uncovered amino acid substitution p.Cys113Tyr in SLC22A4, a transporter of organic cations, cosegregating with HL in FT13 and therefore the cause of ARNSHL DFNB60. We also screened a cohort of small Tunisian HL families and uncovered an additional deaf proband of consanguineous parents that is homozygous for p.Cys113Tyr carried by the same microsatellite marker haplotype as in FT13, indicating that this mutation is ancestral. Using immunofluorescence, we found that Slc22a4 is expressed in stria vascularis (SV) endothelial cells of rodent cochlea and targets their apical plasma membrane. We also found Slc22a4 transcripts in our RNA-seq library from purified primary culture of mouse SV endothelial cells. Interestingly, p.Cys113Tyr mutation affects the trafficking of the transporter and severely alters ergothioneine uptake. We conclude that SLC22A4 is an organic cation transporter of the SV endothelium that is essential for hearing, and its mutation causes DFNB60 form of HL.

Keywords

Hearing Loss Hair Cell Carnitine Organic Cation Transporter Apical Plasma Membrane 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We gratefully thank all the subjects in this study for their collaboration. This study was supported by the National Institutes of Health [Grant Numbers R01DC005575, 2P50DC000422-Sub-Project 6432 and R01DC012115] to X.Z.L., the National Institutes of Health [Grant Number R21DC009879], the University of Miami Provost Research Award, and the College of Arts and Sciences Gabelli Fellowship to Z.L., the National Institutes of Health [Grant Numbers R21DC012398 and R01DC010844] to X.R.S., and the National Institutes of Health [Grant Number R01DC009645] to M.T. This work was also supported by funds from the International Centre for Genetic Engineering and Biotechnology (ICGEB) and the Ministry of Higher Education and Research of Tunisia to S.M., and by a Grant-in-Aid of Scientific Research (B) from the Japanese Ministry of Education, Science and Culture [Grant Number 15H04664] and a Grant from Hoansha Foundation (3-1-8 Dosho-machi, Chuo-ku, Osaka 541-0045, Japan) to Y.K.

Compliance with ethical standards

Conflict of interest

The authors have declared that no competing financial interests exist.

Ethics approval

This study was approved by the institutional review boards (IRB) of the University of Miami and the ethical committee of the University Hospital of Sfax, Tunisia.

Patient consent

Parental consents obtained.

References

  1. Ben Arab S, Masmoudi S, Beltaief N, Hachicha S, Ayadi H (2004) Consanguinity and endogamy in Northern Tunisia and its impact on non-syndromic deafness. Genet Epidemiol 27:74–79CrossRefPubMedGoogle Scholar
  2. Bleasby K, Castle JC, Roberts CJ, Cheng C, Bailey WJ, Sina JF, Kulkarni AV, Hafey MJ, Evers R, Johnson JM et al (2006) Expression profiles of 50 xenobiotic transporter genes in humans and pre-clinical species: a resource for investigations into drug disposition. Xenobiotica 36:963–988CrossRefPubMedGoogle Scholar
  3. Chakchouk I, Ben Said M, Jbeli F, Benmarzoug R, Loukil S, Smeti I, Chakroun A, Gibriel AA, Ghorbel A, Hadjkacem H et al (2015) NADf chip, a two-color microarray for simultaneous screening of multigene mutations associated with hearing impairment in North African Mediterranean countries. J Mol Diagn 17:155–161CrossRefPubMedGoogle Scholar
  4. Cheah IK, Ong RL, Gruber J, Yew TS, Ng LF, Chen CB, Halliwell B (2013) Knockout of a putative ergothioneine transporter in Caenorhabditis elegans decreases lifespan and increases susceptibility to oxidative damage. Free Radic Res 47:1036–1045CrossRefPubMedGoogle Scholar
  5. Clerici WJ, Hensley K, DiMartino DL, Butterfield DA (1996) Direct detection of ototoxicant-induced reactive oxygen species generation in cochlear explants. Hear Res 98:116–124CrossRefPubMedGoogle Scholar
  6. Dehne N, Lautermann J, ten Cate WJ, Rauen U, de Groot H (2000) In vitro effects of hydrogen peroxide on the cochlear neurosensory epithelium of the guinea pig. Hear Res 143:162–170CrossRefPubMedGoogle Scholar
  7. Farthing CA, Sweet DH (2014) Expression and function of organic cation and anion transporters (SLC22 family) in the CNS. Curr Pharm Des 20:1472–1486CrossRefPubMedGoogle Scholar
  8. Forge A, Schacht J (2000) Aminoglycoside antibiotics. Audiol Neurootol 5:3–22CrossRefPubMedGoogle Scholar
  9. Grati M, Kachar B (2011) Myosin VIIa and sans localization at stereocilia upper tip-link density implicates these Usher syndrome proteins in mechanotransduction. Proc Natl Acad Sci USA 108:11476–11481CrossRefPubMedPubMedCentralGoogle Scholar
  10. Grati M, Aggarwal N, Strehler EE, Wenthold RJ (2006) Molecular determinants for differential membrane trafficking of PMCA1 and PMCA2 in mammalian hair cells. J Cell Sci 119:2995–3007CrossRefPubMedGoogle Scholar
  11. Grati M, Chakchouk I, Ma Q, Bensaid M, Desmidt A, Turki N, Yan D, Baanannou A, Mittal R, Driss N et al (2015) A missense mutation in DCDC2 causes human recessive deafness DFNB66, likely by interfering with sensory hair cell and supporting cell cilia length regulation. Hum Mol Genet 24:2482–2491CrossRefPubMedGoogle Scholar
  12. Gründemann D, Harlfinger S, Golz S, Geerts A, Lazar A, Berkels R, Jung N, Rubbert A, Schömig E (2005) Discovery of the ergothioneine transporter. Proc Natl Acad Sci USA 102:5256–5261CrossRefPubMedPubMedCentralGoogle Scholar
  13. Hashimoto N, Suzuki F, Tamai I, Nikaido H, Kuwajima M, Hayakawa J, Tsuji A (1998) Gene-dose effect on carnitine transport activity in embryonic fibroblasts of JVS mice as a model of human carnitine transporter deficiency. Biochem Pharmacol 55:1729–1732CrossRefPubMedGoogle Scholar
  14. Horiuchi M, Kobayashi K, Yamaguchi S, Shimizu N, Koizumi T, Nikaido H, Hayakawa J, Kuwajima M, Saheki T (1994) Primary defect of juvenile visceral steatosis (jvs) mouse with systemic carnitine deficiency is probably in renal carnitine transport system. Biochim Biophys Acta 1226:25–30CrossRefPubMedGoogle Scholar
  15. Kato Y, Kubo Y, Iwata D, Kato S, Sudo T, Sugiura T, Kagaya T, Wakayama T, Hirayama A, Sugimoto M et al (2010) Gene knockout and metabolome analysis of carnitine/organic cation transporter OCTN1. Pharm Res 27:832–840CrossRefPubMedGoogle Scholar
  16. Kawasaki Y, Kato Y, Sai Y, Tsuji A (2004) Functional characterization of human organic cation transporter OCTN1 single nucleotide polymorphisms in the Japanese population. J Pharm Sci 93:2920–2926CrossRefPubMedGoogle Scholar
  17. Lautermann J, Crann SA, McLaren J, Schacht J (1997) Glutathione-dependent antioxidant systems in the mammalian inner ear: effects of aging, ototoxic drugs and noise. Hear Res 114:75–82CrossRefPubMedGoogle Scholar
  18. Neng L, Zhang W, Hassan A, Zemla M, Kachelmeier A, Fridberger A, Auer M, Shi X (2013) Isolation and culture of endothelial cells, pericytes and perivascular resident macrophage-like melanocytes from the young mouse ear. Nat Protoc 8:709–720CrossRefPubMedPubMedCentralGoogle Scholar
  19. Nezu J, Tamai I, Oku A, Ohashi R, Yabuuchi H, Hashimoto N, Nikaido H, Sai Y, Koizumi A, Shoji Y et al (1999) Primary systemic carnitine deficiency is caused by mutations in a gene encoding sodium ion-dependent carnitine transporter. Nat Genet 21:91–94CrossRefPubMedGoogle Scholar
  20. Pelis RM, Wright SH (2014) SLC22, SLC44, and SLC47 transporters–organic anion and cation transporters: molecular and cellular properties. Curr Top Membr 73:233–261CrossRefPubMedGoogle Scholar
  21. Peltekova VD, Wintle RF, Rubin LA, Amos CI, Huang Q, Gu X, Newman B, Van Oene M, Cescon D, Greenberg G et al (2004) Functional variants of OCTN cation transporter genes are associated with Crohn disease. Nat Genet 36:471–475CrossRefPubMedGoogle Scholar
  22. Shen J, Scheffer DI, Kwan KY, Corey DP (2015) SHIELD: an integrative gene expression database for inner ear research. Database 2015:1–9CrossRefGoogle Scholar
  23. Tamai I (2013) Pharmacological and pathophysiological roles of carnitine/organic cation transporters (OCTNs: SLC22A4, SLC22A5 and Slc22a21). Biopharm Drug Dispos 34:29–44CrossRefPubMedGoogle Scholar
  24. Tamai I, Ohashi R, Nezu J, Yabuuchi H, Oku A, Shimane M, Sai Y, Tsuji A (1998) Molecular and functional identification of sodium ion-dependent, high affinity human carnitine transporter OCTN2. J Biol Chem 273:20378–20382CrossRefPubMedGoogle Scholar
  25. Taubert D, Jung N, Goeser T, Schömig E (2009) Increased ergothioneine tissue concentrations in carriers of the Crohn’s disease risk-associated 503F variant of the organic cation transporter OCTN1. Gut 58:312–314CrossRefPubMedGoogle Scholar
  26. Tokuhiro S, Yamada R, Chang X, Suzuki A, Kochi Y, Sawada T, Suzuki M, Nagasaki M, Ohtsuki M, Ono M et al (2003) An intronic SNP in a RUNX1 binding site of SLC22A4, encoding an organic cation transporter, is associated with rheumatoid arthritis. Nat Genet 35:341–348CrossRefPubMedGoogle Scholar
  27. Trapnell C, Roberts A, Goff L, Pertea G, Kim D, Kelley DR, Pimentel H, Salzberg SL, Rinn JL, Pachter L (2012) Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nat Protoc 7:562–578CrossRefPubMedPubMedCentralGoogle Scholar
  28. Yabuuchi H, Tamai I, Nezu J, Sakamoto K, Oku A, Shimane M, Sai Y, Tsuji A (1999) Novel membrane transporter OCTN1 mediates multispecific, bidirectional, and pH-dependent transport of organic cations. J Pharmacol Exp Ther 289:768–773PubMedGoogle Scholar
  29. Yamane H, Nakai Y, Takayama M, Iguchi H, Nakagawa T, Kojima A (1995) Appearance of free radicals in the guinea pig inner ear after noise-induced acoustic trauma. Eur Arch Otorhinolaryngol 252:504–508CrossRefPubMedGoogle Scholar
  30. Yasunaga S, Grati M, Cohen-Salmon M, El-Amraoui A, Mustapha M, Salem N, El-Zir E, Loiselet J, Petit C (1999) A mutation in OTOF, encoding otoferlin, a FER-1-like protein, causes DFNB9, a nonsyndromic form of deafness. Nat Genet 21:363–369CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Mariem Ben Said
    • 1
  • M’hamed Grati
    • 2
  • Takahiro Ishimoto
    • 3
  • Bing Zou
    • 2
  • Imen Chakchouk
    • 1
  • Qi Ma
    • 2
  • Qi Yao
    • 2
    • 4
  • Bouthaina Hammami
    • 5
  • Denise Yan
    • 2
  • Rahul Mittal
    • 2
  • Noritaka Nakamichi
    • 3
  • Abdelmonem Ghorbel
    • 5
  • Lingling Neng
    • 6
  • Mustafa Tekin
    • 2
    • 7
  • Xiao Rui Shi
    • 6
  • Yukio Kato
    • 3
  • Saber Masmoudi
    • 1
    Email author
  • Zhongmin Lu
    • 4
  • Mounira Hmani
    • 1
  • Xuezhong Liu
    • 2
    • 7
    • 8
    Email author
  1. 1.Laboratoire Procédés de Criblage Moléculaire et Cellulaire, Centre de Biotechnologie de SfaxUniversité de SfaxSfaxTunisia
  2. 2.Department of Otolaryngology (D-48)University of Miami Miller School of MedicineMiamiUSA
  3. 3.Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health SciencesKanazawa UniversityKanazawaJapan
  4. 4.Department of BiologyUniversity of MiamiMiamiUSA
  5. 5.Service OtorhinolaryngologieHôpital Universitaire Habib BourguibaSfaxTunisia
  6. 6.Oregon Hearing Research Center, Department of Otolaryngology/Head and Neck SurgeryOregon Health and Science UniversityPortlandUSA
  7. 7.Dr. John T. Macdonald Foundation Department of Human Genetics, and John P. Hussman Institute for Human GenomicsUniversity of MiamiMiamiUSA
  8. 8.Department of Otolaryngology, Xiangya HospitalCentral South UniversityChangshaChina

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