The actin cytoskeleton plays important roles in various cell structures and activities. The formation and function of the actin cytoskeleton require the actin-based motors, myosins, which comprise a large superfamily of adenosine triphosphatases (ATPases). Utilizing the energy released from ATP hydrolysis, myosins move along actin filaments to generate motility or force. Each myosin contains a highly conserved N-terminal motor (or head) domain, a central neck domain and a C-terminal tail region. The motor domain contains ATPase activity that catalyzes ATP hydrolysis and regulates actin-binding. The neck domain binds light chains or calmodulins, and acts as a lever arm for force transduction. The C-terminal tail region is the most divergent among the different myosins and contains various domains that mediate oligomerization or cargo interactions.
Myosins form a large and diverse protein superfamily, whose members are grouped into more than 30 distinct classes. Myosin II (MYO2), the first myosin discovered, is designated as a “conventional myosin”, and the other myosins are considered as “unconventional myosins”. The conventional myosin is found in muscle cells and the contractile ring in non-muscle cells, forming bipolar thick filaments via tail-mediated oligomerization that pull actin filaments to produce contraction. The unconventional myosins, however, do not form bipolar thick filaments. Instead, their tails mediate binding to the plasma membrane or cargo proteins. The unconventional myosins are implicated in diverse cellular processes, such as organelle trafficking, F-actin organization and cell movement, among others.
Myosin tail homology 4 - protein 4.1, ezrin, radixin, and moesin (MyTH4-FERM) myosins are a group of unconventional myosins involved in actin-based finger-like protrusions such as filopodia, microvilli, and stereocilia. Known MyTH4-FERM myosins include MYO7A, MYO7B, MYO10, and MYO15A, all of which contain the MyTH4-FERM domain in their C-terminal tail regions (Fig. 1). The MyTH4 and FERM domains form a structural and functional supramodule, the MyTH4 domain mediating microtubule binding and the FERM domain mediating cargo interactions.[7–9] MYO10 has one MyTH4-FERM domain after a single α-helix (SAH) domain, a proline, glutamate, serine, threonine peptide (PEST) sequence and several Pleckstrin homology (PH) domains. MYO7A, MYO7B, and MYO15A have 2 tandem MyTH4-FERM domains separated by an SRC homology 3 (SH3) domain. MYO15A also contains a PDZ-binding interface (PBI) at the C-terminal end that mediates interaction with the PDZ (PSD95, Dlg1, and ZO-1) domain.
In this review, we will focus on the unconventional myosin MYO15A. MYO15A is localized at the tips of stereocilia in inner ear hair cells and plays important roles in development and maintenance of stereocilia. We will discuss the expression pattern of the MYO15A gene in the inner ear, the biochemical properties of the MYO15A protein, the association of MYO15A mutations with hearing loss and our present understanding of how MYO15A regulates stereocilia development and maintenance.
Database Search Strategy
An electronic search of the PubMed database was performed to identify relevant publications from 1945 to 2019 using the following “myosin 15A” OR “myosin XVA” OR “myo15a”. The results were further screened by title and abstract to only present studies describing the expression; properties; interactions; or functions of MYO15A. Lastly, the authors screened the reference list of included publications to identify other potentially relevant publications.
MYO15A/Myo15a gene expression
Both human MYO15A and mouse Myo15a genes contain 66 exons, encoding a large protein of up to nearly 400 kDa. Exon 2 of the MYO15A/Myo15a gene is subjected to alternative splicing, which gives rise to MYO15A isoforms of different lengths. The short MYO15A isoform (MYO15A-S) contains a N-terminal motor domain, a central neck domain and a C-terminal tail region. The long MYO15A isoform (MYO15A-L) has an additional large N-terminal extension of unknown function before the motor domain (Fig. 1). Moreover, alternative splicing was observed in exons 8 and 26, which are 6 nucleotides (nt) and 216 nt/189 nt (human/mouse) long, respectively. Inclusion of exon 8 affects the motor domain, while inclusion of exon 26 affects the neck domain, whose biological significance remains undefined.
In situ hybridization using a probe for both long and short isoforms showed that Myo15a transcripts are first expressed in the developing inner ear at embryonic day 13.5 (E13.5). At E17.5, Myo15a expression is restricted to the sensory epithelium in both the vestibula and the cochlea. At postnatal day 8 (P8), Myo15a transcripts are only detected in the hair cells but not in the supporting cells. Similar results were obtained when a long isoform-specific probe was used. The specific expression pattern of Myo15a transcripts in the inner ear, especially in the hair cells, suggests that MYO15A might play important roles in the development and/or function of the hair cells.
Furthermore, quantitative PCR (qPCR) showed that Myo15a transcripts exhibit isoform switching during development in the mouse cochlea.Myo15a-l transcripts become progressively more abundant during development, increasing about 9-fold from P0 to P21. In contrast, expression of Myo15a-s transcripts is significantly decreased about 21-fold from P0 to P6. Meanwhile, the total amount of Myo15a transcripts remains stable during development. The developmentally isoform switching suggests that the different MYO15A isoforms play different roles in the hair cells.
Biochemical properties of MYO15A protein
The biochemical properties of MYO15A have been investigated using its purified motor domain plus the neck region. The results showed that MYO15A is a plus end-directed myosin that moves rapidly along the actin filaments. The duty ratio is an important property for myosins, defined as the fraction of the ATPase cycle that myosin spends strongly associated with F-actin. MYO15A is a high-duty ratio motor, suggesting that it can move along F-actin processively. In vitro experiments also showed that heat shock protein 90 (HSP90) and its co-chaperone UNC-45B could bind to MYO15A and promote its folding.
The neck region of MYO15A contains 3 predicted light-chain binding sites (IQ motifs). The first and second IQ motifs bind to regulatory light chain (RLC) and essential light chain (ELC), respectively, while the third IQ motif seems to be unoccupied. Calmodulin could compete with ELC for binding to the second IQ motif, although with low affinity. MYO15A does not have a predicted coiled-coil sequence and, at present, it is unknown whether it could dimerize in vivo.
The C-terminal tail of MYO15A contains two tandem MyTH4-FERM domains separated by an SH3 domain, as well as a PBI at the C-terminal end. It has been suggested that MYO15A binds to the C terminus of the actin-regulatory protein, epidermal growth factor receptor kinase substrate 8 (EPS8), possibly via its MyTH4-FERM domain. MYO15A also binds to the PDZ domains of scaffold protein whirlin (WHRN) via its C-terminal MyTH4-FERM and PBI.[15,16] In summary, MYO15A is a plus end-directed, high-duty ratio motor, and interacts with RLC, ELC, calmodulin, EPS8, and WHRN via its multiple domains. Mutations that affect these domains destroy the function of MYO15A and lead to hearing loss, which will be discussed in the next section.
Mutations in the MYO15A/Myo15a gene cause hearing loss in humans and rodents
In 1993, the gene associated with congenital, recessive deafness DFNB3 was mapped to the pericentromeric region of chromosome 17, which was subsequently refined to 17p11.2.[17,18] In 1998, mutations in the human MYO15A gene were identified as responsible for DFNB3. Since then, more than 70 mutations in MYO15A gene have been identified in patients with hearing loss (Fig. 2).[20–44] These mutations cover the full length of MYO15A, affecting every important domain and motif.
Animal models containing Myo15a gene mutations have also been identified. Shaker-2 (sh2) is a recessive mouse mutant with hearing and balancing problems and has been suggested as the mouse model of DFNB3. In 1998, mutation in the Myo15a gene was identified as responsible for the phenotypes in sh2 mice.[45,46] Another mouse line, shaker-2j (sh2j), has similar phenotypes as sh2, and mutation in the Myo15a gene was identified as the causative gene. The mutations in sh2 and sh2j mice affect the motor and tail domains of MYO15A, respectively, suggesting that both regions are critical for the function of MYO15A in hair cells. A deletion in the Myo15a gene that encodes part of the motor domain was then identified in the spontaneous deaf kuru2 mice. Recently, a knock-in mouse line was established by introducing an E1086X nonsense mutation into the Myo15a gene, mimicking the E1105X mutation in DFNB3 patients.[12,21] Because this mutation lies in exon 2, it only affects MYO15A-L, but leaves MYO15A-S unaffected. The resultant Myo15aΔN/ΔN mice show less severe hearing loss compared with sh2 or sh2j mice and normal vestibular function. Lastly, a spontaneous mutation L3157P in the second MyTH4 domain of MYO15A was identified in the circling-2 (ci2) rats that possess hearing, balancing, and vision deficits. These animal models provide valuable tools for the investigation into the function of MYO15A in hearing.
MYO15A regulates the development and maintenance of stereocilia
Immunostaining with an antibody against common epitopes present in both isoforms, PB48, showed that MYO15A is localized at the tips of all stereocilia in both OHCs and IHCs (Fig. 3A and B).[12,49] MYO15A immunoreactivity was first detected at the stereociliary tips of mouse E14.5 vestibular hair cells and E18.5 cochlear hair cells, coinciding with the onset of the staircase architecture formation of the stereocilia. The two known MYO15A binding partners, EPS8 and WHRN, are also localized at the tips of stereocilia.[14–16] MYO15A, EPS8, and WHRN form a tripartite complex at the tips of stereocilia and regulate stereocilia elongation. The same antibody, PB48, was also used in immunostaining of Myo15a△N/△N mice that lack MYO15A-L, which revealed that MYO15A-S is localized at the tips of the tallest row stereocilia in both OHCs and IHCs (Fig. 3C and D). The localization of MYO15A-L was examined by performing immunostaining of normal control mice with PB888, an antibody specific to the N-terminal extension part. The results suggested that MYO15A-L is localized at the tips of the shorter row stereocilia in IHCs but at the tips of all stereocilia in OHCs (Fig. 3E and F).
The role of MYO15A in the development and/or maintenance of stereocilia was further investigated using Myo15a mutant mice. In sh2/sh2 or sh2j/sh2j mice that lack both functional MYO15A isoforms, the stereocilia of cochlear and vestibular hair cells are extremely short.[11,45] Hair bundles in their IHCs also show supernumerary stereocilia rows. Similar stereocilia abnormalities were also observed in Whrn- or Eps8-deficient mice.[14,50] However, overexpressed MYO15A-S in normal control hair cells has no effect on the elongation of developing stereocilia. Interestingly, the effect of Myo15a disruption on stereocilia development differs between IHCs and OHCs. In sh2/sh2 IHCs, all the stereocilia are equally short, and no obliquely oriented tip-links are observed. However, the short stereocilia of sh2/sh2 OHCs still retain a staircase arrangement with prominent tip-links. It has also been shown that fast adaptation and Ca2+ sensitivity of mechanical-electrical transduction (MET) are affected in IHCs but not OHCs of sh2/sh2 mice.
In the Myo15a△N/△N mice that lack MYO15A-L, the development of the staircase architecture of stereocilia is unaffected by P10, suggesting that MYO15A-L is not required for stereocilia development. However, from P32 onwards in Myo15a△N/△N hair cells, the second row stereocilia are significantly shorter, and the third row stereocilia are almost completely resorbed, whereas the tallest row stereocilia are largely unaffected, suggesting that MYO15A-L is indispensable for the postnatal maintenance of the shorter row stereocilia. Further examination of the ultrastructure of Myo15a△N/△N shorter row stereocilia at P8 revealed exaggerated and over-elongated prolate tips filled with actin filaments, suggesting that actin polymerization was dysregulated at this position. It is worth noticing that the localization of WHRN and EPS8 is unaffected in the Myo15a△N/△N mice, suggesting that MYO15A-S is sufficient to traffic these two proteins to the tips of stereocilia. The sensitivity of MET was also affected in the Myo15a△N/△N IHCs but not OHCs. In summary, the present data suggest that MYO15A-S and MYO15A-L are localized at the tips of stereocilia of different rows, and play different but important roles in the development and maintenance of stereocilia (Fig. 3G and H).
Conclusion and perspectives
Research over the last 2 decades suggest that the unconventional myosin MYO15A is localized at the tips of stereocilia and regulates stereocilia development/maintenance. The MYO15A gene is subjected to alternative splicing, which gives rise to different MYO15A isoforms with slightly different localization and functions. MYO15A-S is localized at the tips of the tallest row stereocilia and is necessary for early stereocilia development. However, MYO15A-L is localized at the tips of the shorter row stereocilia in IHCs, and is indispensable for the postnatal maintenance of the shorter row stereocilia. At present it remains unclear how the different localization and function of the 2 MYO15A isoforms are regulated. The N-terminal extension part that is unique to MYO15A-L must be important for this discrepancy. Therefore, identification of proteins that specifically bind to this N-terminal extension will certainly help answer this question.
At the tips of the tallest row stereocilia, MYO15A-S interacts with WHRN and EPS8, forming a so-called stereocilia tip complex. This complex also contains other components such as GPSM2 and Gαi3.[52,53] There is evidence suggesting that MYO15A-EPS8 forms the core of this complex and is transported to the stereocilia tips first, then WHRN-GPSM2-Gαi3 is added to the pre-existing MYO15A-EPS8 stereocilia tip complex. Mutations that affect any component in this complex cause similar stereocilia elongation phenotypes, further supporting the role of this complex in stereocilia development. At present how this complex regulates the length of the stereocilia is still unknown and awaits further investigation.
MYO15A-L is localized at the tips of shorter row stereocilia in IHCs, where actin capping protein subunit B2 (CAPZB2), twinfilin 2 (TWF2) and EPS8-like 2 (EPS8L2) also reside.[54–57] Mice lacking CAPZB2 or EPS8L2 show stereocilia maintenance abnormality similar to that observed in Myo15a△N/△N mice.[56,57] At present it is unknown whether all these proteins can bind to each other and form a protein complex, although CAPZB2 has been shown to bind TWF2. The MET complex also localizes at the tips of shorter stereocilia, near the lower end of tip links.[58–62] It remains to be tested whether these proteins have connections with the MET complex. However, the normal amplitude of MET currents in Myo15a or Eps8l2 mutant mice suggests that neither MYO15A nor EPS8L2 plays an indispensable role in MET.[12,51,56] Further investigations are warranted to learn more about all these proteins localized at the tips of shorter stereocilia.
Different expression patterns and roles of MYO15A isoforms in the stereocilia suggest different intervention strategies for patients with different MYO15A mutations. Adeno-associated viruses were proven to efficiently deliver genes into inner ear cells.[63–65] This approach has been successfully applied in the treatment of hearing loss in several mouse models.[66–71] Furthermore, the powerful gene editing techniques provide a promising therapeutic strategy towards hearing loss.[72,73] Recently this strategy was employed to successfully improve the auditory function of the Tmc1 mutant deaf mice.[74,75] For the potential treatment of MYO15A-related hearing loss, a bacterial artificial chromosome containing full-length Myo15a gene has been shown to rescue the deafness phenotype in sh2 mice. Compared to MYO15A-S, MYO15A-L is not required for the stereocilia until a later developmental stage, which gives patients with mutations in MYO15A exon 2 a broader time window for intervention. Further investigations into the function and regulation of MYO15A will certainly help to develop more efficient treatments of related patients.
HD, NL and ZX retrieved the literature and wrote the manuscript. All the authors approved the final version of the paper.
Our laboratory is supported by the National Key Basic Research Program of China (No. 2018YFC1003600), the National Natural Science Foundation of China (No. 81771001), Shandong Provincial Key Laboratory of Animal Cell and Developmental Biology (No. SPKLACDB-2019000), and the Fundamental Research Funds of Shandong University (No. 2018JC025).
Conflicts of interest
The authors declare that they have no conflicts of interest.
. Masters TA, Kendrick-Jones J, Buss F. Myosins
: domain organisation, motor properties, physiological roles and cellular functions. Handb Exp Pharmacol 2017;235:77–122.
. Kendrick-Jones J, Buss F. Editorial overview: myosins
in review. Traffic 2016;17:819–821.
. Reiser PJ. Current understanding of conventional and novel co-expression patterns of mammalian sarcomeric myosin heavy chains and light chains. Arch Biochem Biophys 2019;662:129–133.
. Batters C, Veigel C. Mechanics and activation of unconventional myosins
. Traffic 2016;17:860–871.
. Fili N, Toseland CP. Unconventional myosins
: how regulation meets function. Int J Mol Sci 2019;21:E67.
. Weck ML, Grega-Larson NE, Tyska MJ. MyTH4-FERM myosins
in the assembly and maintenance of actin-based protrusions. Curr Opin Cell Biol 2017;44:68–78.
. Hirano Y, Hatano T, Takahashi A, et al. Structural basis of cargo recognition by the myosin-X MyTH4-FERM domain. EMBO J 2011;30:2734–2747.
. Wei Z, Yan J, Lu Q, et al. Cargo recognition mechanism of myosin X revealed by the structure of its tail MyTH4-FERM tandem in complex with the DCC P3 domain. Proc Natl Acad Sci U S A 2011;108:3572–3577.
. Wu L, Pan L, Wei Z, et al. Structure of MyTH4-FERM domains in myosin VIIa tail bound to cargo. Science 2011;331:757–760.
. Liang Y, Wang A, Belyantseva IA, et al. Characterization of the human and mouse unconventional myosin XV genes responsible for hereditary deafness DFNB3 and shaker 2. Genomics 1999;61:243–258.
. Anderson DW, Probst FJ, Belyantseva IA, et al. The motor and tail regions of myosin XV are critical for normal structure and function of auditory and vestibular hair cells
. Hum Mol Genet 2000;9:1729–1738.
. Fang Q, Indzhykulian AA, Mustapha M, et al. The 133-kDa N-terminal domain enables myosin 15 to maintain mechanotransducing stereocilia
and is essential for hearing. Elife 2015;4:e08627.
. Bird JE, Takagi Y, Billington N, et al. Chaperone-enhanced purification of unconventional myosin 15, a molecular motor specialized for stereocilia
protein trafficking. Proc Natl Acad Sci U S A 2014;111:12390–12395.
. Manor U, Disanza A, Grati MH, et al. Regulation of stereocilia
length by myosin XVa and whirlin depends on the actin-regulatory protein Eps8. Curr Biol 2011;21:167–172.
. Belyantseva IA, Boger ET, Naz S, et al. Myosin-XVa is required for tip localization of whirlin and differential elongation of hair-cell stereocilia
. Nat Cell Biol 2005;7:148–156.
. Delprat B, Michel V, Goodyear R, et al. Myosin XVa and whirlin, two deafness gene products required for hair bundle growth, are located at the stereocilia
tips and interact directly. Hum Mol Genet 2005;14:401–410.
. Friedman TB, Liang Y, Weber JL, et al. A gene for congenital, recessive deafness DFNB3 maps to the pericentromeric region of chromosome 17. Nat Genet 1995;9:86–91.
. Liang Y, Wang A, Probst FJ, et al. Genetic mapping refines DFNB3 to 17p11.2, suggests multiple alleles of DFNB3, and supports homology to the mouse model shaker-2. Amer J Hum Genet 1998;62:904–915.
. Wang A, Liang Y, Fridell RA, et al. Association of unconventional myosin MYO15 mutations with human nonsyndromic deafness DFNB3. Science 1998;280:1447–1451.
. Liburd N, Ghosh M, Riazuddin S, et al. Novel mutations of MYO15A
associated with profound deafness in consanguineous families and moderately severe hearing loss in a patient with Smith-Magenis syndrome. Hum Genet 2001;109:535–541.
. Nal N, Ahmed ZM, Erkal E, et al. Mutational spectrum of MYO15A
: the large N-terminal extension of myosin XVA is required for hearing. Hum Mutat 2007;28:1014–1019.
. Kalay E, Uzumcu A, Krieger E, et al. MYO15A
(DFNB3) mutations in Turkish hearing loss families and functional modeling of a novel motor domain mutation. Am J Med Genet A 2007;143A:2382–2389.
. Lezirovitz K, Pardono E, de Mello Auricchio MTB, et al. Unexpected genetic heterogeneity in a large consanguineous Brazilian pedigree presenting deafness. Eur J Hum Genet 2008;16:89–96.
. Shearer AE, Hildebrand MS, Webster JA, et al. Mutations in the first MyTH4 domain of MYO15A
are a common cause of DFNB3 hearing loss. Laryngoscope 2009;119:727–733.
. Belguith H, Aifa-Hmani M, Dhouib H, et al. Screening of the DFNB3 locus: identification of three novel mutations of MYO15A
associated with hearing loss and further suggestion for two distinctive genes on this locus. Genet Test Mol Biomarkers 2009;13:147–151.
. Shahin H, Walsh T, Rayyan AA, et al. Five novel loci for inherited hearing loss mapped by SNP-based homozygosity profiles in Palestinian families. Eur J Hum Genet 2010;18:407–413.
. Cengiz FB, Duman D, Sirmaci A, et al. Recurrent and private MYO15A
mutations are associated with deafness in the Turkish population. Genet Test Mol Biomarkers 2010;14:543–550.
. Brownstein Z, Friedman LM, Shahin H, et al. Targeted genomic capture and massively parallel sequencing to identify genes for hereditary hearing loss in Middle Eastern families. Genome Biol 2011;12:R89.
. Imtiaz F, Taibah K, Ramzan K, et al. A comprehensive introduction to the genetic basis of non-syndromic hearing loss in the Saudi Arabian population. BMC Med Genet 2011;12:91.
. Duman D, Sirmaci A, Cengiz FB, et al. Screening of 38 genes identifies mutations in 62% of families with nonsyndromic deafness in Turkey. Genet Test Mol Biomarkers 2011;15:29–33.
. Diaz-Horta O, Duman D, Foster J 2nd, et al. Whole-exome sequencing efficiently detects rare mutations in autosomal recessive nonsyndromic hearing loss. PLoS One 2012;7:e50628.
. Fattahi Z, Shearer AE, Babanejad M, et al. Screening for MYO15A
gene mutations in autosomal recessive nonsyndromic, GJB2 negative Iranian deaf population. Am J Med Genet A 2012;158A:1857–1864.
. Gao X, Zhu QY, Song YS, et al. Novel compound heterozygous mutations in the MYO15A
gene in autosomal recessive hearing loss identified by whole-exome sequencing. J Transl Med 2013;11:284–1284.
. Miyagawa M, Nishio SY, Ikeda T, et al. Massively parallel DNA sequencing successfully identifies new causative mutations in deafness genes in patients with cochlear implantation and EAS. PLoS One 2013;8:e75793.
. Yang T, Wei X, Chai Y, et al. Genetic etiology study of the non-syndromic deafness in Chinese Hans by targeted next-generation sequencing. Orphanet J Rare Dis 2013;8:85.
. Brownstein Z, Abu-Rayyan A, Karfunkel-Doron D, et al. Novel myosin mutations for hereditary hearing loss revealed by targeted genomic capture and massively parallel sequencing. Eur J Hum Genet 2014;22:768–775.
. Miyagawa M, Nishio S-Y, Hattori M, et al. Mutations in the MYO15A
gene are a significant cause of nonsyndromic hearing loss: massively parallel DNA sequencing-based analysis. Ann Otol Rhinol Laryngol 2015;124 (Suppl 1):158S–168S.
. Xia H, Huang X, Guo Y, et al. Identification of a novel MYO15A
mutation in a Chinese family with autosomal recessive nonsyndromic hearing loss. PLoS One 2015;10:e0136306.
. Li W, Guo L, Li Y, et al. A novel recessive truncating mutation in MYO15A
causing prelingual sensorineural hearing loss. Int J Pediatr Otorhinolaryngol 2016;81:92–95.
. Reiisi S, Tabatabaiefar MA, Sanati MH, et al. Screening of DFNB3 in Iranian families with autosomal recessive non-syndromic hearing loss reveals a novel pathogenic mutation in the MyTh4 domain of the MYO15A
gene in a linked family. Iran J Basic Med Sci 2016;19:772–778.
. Salime S, Charif M, Bousfiha A, et al. Homozygous mutations in PJVK and MYO15A
genes associated with non-syndromic hearing loss in Moroccan families. Int J Pediatr Otorhinolaryngol 2017;101:25–29.
. Zhou H, Kuermanhan A, Zhang Z, et al. Identification of a novel homozygous mutation in the MYO15A
gene in a Kazakh family with non-syndromic hearing loss. Int J Pediatr Otorhinolaryngol 2019;125:128–132.
. Mehregan H, Mohseni M, Jalalvand K, et al. Novel mutations in MYTH4-FERM domains of myosin 15 are associated with autosomal recessive nonsyndromic hearing loss. Int J Pediatr Otorhinolaryngol 2019;117:115–126.
. Akbariazar E, Vahabi A, Abdi Rad I. Report of a novel splicing mutation in the MYO15A
gene in a patient with sensorineural hearing loss and spectrum of the MYO15A
mutations. Clin Med Insights Case Rep 2019;12:1179547619871907.
. Probst FJ, Fridell RA, Raphael Y, et al. Correction of deafness in shaker-2 mice by an unconventional myosin in a BAC transgene. Science 1998;280:1444–1447.
. Wakabayashi Y, Takahashi Y, Kikkawa Y, et al. A novel type of myosin encoded by the mouse deafness gene shaker-2. Biochem Biophys Res Commun 1998;248:655–659.
. Watanabe M, Akiyama N, Manome Y, et al. Spontaneous mutant ICR kuru2 might be another shaker-2 deaf mouse. In Vivo 2012;26:787–791.
. Held N, Smits BMG, Gockeln R, et al. A mutation in Myo15 leads to Usher-like symptoms in LEW/Ztm-ci2 rats. PLoS One 2011;6:e15669.
. Belyantseva IA, Boger ET, Friedman TB. Myosin XVa localizes to the tips of inner ear
sensory cell stereocilia
and is essential for staircase formation of the hair bundle. Proc Natl Acad Sci U S A 2003;100:13958–13963.
. Holme RH, Kiernan BW, Brown SDM, et al. Elongation of hair cell stereocilia
is defective in the mouse mutant whirler. J Comp Neurol 2002;450:94–102.
. Stepanyan R, Frolenkov GI. Fast adaptation and Ca2+ sensitivity of the mechanotransducer require myosin-XVa in inner but not outer cochlear hair cells
. J Neurosci 2009;29:4023–4034.
. Mauriac SA, Hien YE, Bird JE, et al. Defective Gpsm2/Gα(i3) signalling disrupts stereocilia
development and growth cone actin dynamics in Chudley-McCullough syndrome. Nat Commun 2017;8:14907.
. Tadenev ALD, Akturk A, Devanney N, et al. GPSM2-GNAI specifies the tallest stereocilia
and defines hair bundle row identity. Curr Biol 2019;29:921–934.
. Peng AW, Belyantseva IA, Hsu PD, et al. Twinfilin 2 regulates actin filament lengths in cochlear stereocilia
. J Neurosci 2009;29:15083–15088.
. Rzadzinska AK, Nevalainen EM, Prosser HM, et al. MyosinVIIa interacts with Twinfilin-2 at the tips of mechanosensory stereocilia
in the inner ear
. PLoS One 2009;4:e7097.
. Furness DN, Johnson SL, Manor U, et al. Progressive hearing loss and gradual deterioration of sensory hair bundles in the ears of mice lacking the actin-binding protein Eps8L2. Proc Natl Acad Sci U S A 2013;110:13898–13903.
. Avenarius MR, Krey JF, Dumont RA, et al. Heterodimeric capping protein is required for stereocilia
length and width regulation. J Cell Biol 2017;216:3861–3881.
. Beurg M, Fettiplace R, Nam J-H, et al. Localization of inner hair cell mechanotransducer channels using high-speed calcium imaging. Nat Neurosci 2009;12:553–558.
. Xiong W, Grillet N, Elledge HM, et al. TMHS is an integral component of the mechanotransduction machinery of cochlear hair cells
. Cell 2012;151:1283–1295.
. Zhao B, Wu Z, Grillet N, et al. TMIE is an essential component of the mechanotransduction machinery of cochlear hair cells
. Neuron 2014;84:954–967.
. Kurima K, Ebrahim S, Pan B, et al. TMC1 and TMC2 localize at the site of mechanotransduction in mammalian inner ear
hair cell stereocilia
. Cell Rep 2015;12:1606–1617.
. Li X, Yu X, Chen X, et al. Localization of TMC1 and LHFPL5 in auditory hair cells
in neonatal and adult mice. FASEB J 2019;33:6838–6851.
. Landegger LD, Pan B, Askew C, et al. A synthetic AAV vector enables safe and efficient gene transfer to the mammalian inner ear
. Nat Biotechnol 2017;35:280–284.
. Isgrig K, McDougald DS, Zhu J, et al. AAV2.7m8 is a powerful viral vector for inner ear
gene therapy. Nat Commun 2019;10:427.
. Tan F, Chu C, Qi J, et al. AAV-ie enables safe and efficient gene transfer to inner ear
cells. Nat Commun 2019;10:3733.
. Pan B, Askew C, Galvin A, et al. Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c. Nat Biotechnol 2017;35:264–272.
. Dulon D, Papal S, Patni P, et al. Clarin-1 gene transfer rescues auditory synaptopathy in model of Usher syndrome. J Clin Invest 2018;128:3382–3401.
. Al-Moyed H, Cepeda AP, Jung S, et al. A dual-AAV approach restores fast exocytosis and partially rescues auditory function in deaf otoferlin knock-out mice. EMBO Mol Med 2019;11:e9396.
. Akil O, Dyka F, Calvet C, et al. Dual AAV-mediated gene therapy restores hearing in a DFNB9 mouse model. Proc Natl Acad Sci U S A 2019;116:4496–4501.
. Reisinger E. Dual-AAV delivery of large gene sequences to the inner ear
. Hearing Res 2019;107857.
. Nist-Lund CA, Pan B, Patterson A, et al. Improved TMC1 gene therapy restores hearing and balance in mice with genetic inner ear
disorders. Nat Commun 2019;10:236–1236.
. Rees HA, Liu DR. Base editing: precision chemistry on the genome and transcriptome of living cells. Nat Rev Genet 2018;19:770–788.
. Zhou C, Sun Y, Yan R, et al. Off-target RNA mutation induced by DNA base editing and its elimination by mutagenesis. Nature 2019;571:275–278.
. Zuris JA, Thompson DB, Shu Y, et al. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat Biotechnol 2015;33:73–80.
. Gao X, Tao Y, Lamas V, et al. Treatment of autosomal dominant hearing loss by in vivo delivery of genome editing agents. Nature 2018;553:217–221.