AAV-ie enables safe and efficient gene transfer to inner ear cells

Tan, Fangzhi; Chu, Cenfeng; Qi, Jieyu; Li, Wenyan; You, Dan; Li, Ké; Chen, Xin; Zhang, Weidong; Cheng, Cheng; Liu, Xiaoyi; Qiao, Yong; Su, Bing; He, Shuai; Zhong, Chao; Li, Huawei; Chai, Renjie; Zhong, Guisheng

doi:10.1038/s41467-019-11687-8

Cited by 165 publications

(144 citation statements)

References 38 publications

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“…Viral vectors such as AAV-inner ear, AAV2.7m8, bovine AAV, and adenovirus may be more suitable for selective SC transduction. 9,[19][20][21] It remains unclear why our results appear to diverge from those in prior studies, particularly the observed transduction rates of AAV1 and Anc80 for IHCs and OHCs, respectively, as compared with AAV2. By design, our study leverages a route of administration that differs from routes used in other studies assessing AAV transduction in the cochlea.…”

Section: Discussioncontrasting

confidence: 96%

Hair Cell Transduction Efficiency of Single- and Dual-AAV Serotypes in Adult Murine Cochleae

Omichi

Yoshimura

Shibata

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Gene delivery is a key component for the treatment of genetic hearing loss. To date, a myriad of adeno-associated virus (AAV) serotypes and surgical approaches have been employed to deliver transgenes to cochlear hair cells, but the efficacy of dual transduction remains unclear. Herein, we investigated cellular tropism of single injections of AAV serotype 1 (AAV1), AAV2, AAV8, AAV9, and Anc80L65, and quantitated dual-vector co-transduction rates following co-injection of AAV2 and AAV9 vectors in adult murine cochlea. We used the combined round window membrane and canal fenestration (RWM+CF) injection technique for vector delivery. Single AAV2 injections were most robust and transduced 96.7% ± 1.1% of inner hair cells (IHCs) and 83.9% ± 2.0% of outer hair cells (OHCs) throughout the cochlea without causing hearing impairment or hair cell loss. Dual AAV2 injection co-transduced 96.9% ± 1.7% of IHCs and 65.6% ± 8.95% of OHCs. Together, RWM+CF-injected single or dual AAV2 provides the highest auditory hair cell transduction efficiency of the AAV serotypes we studied. These findings broaden the application of cochlear gene therapy targeting hair cells.

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Section: Discussioncontrasting

confidence: 96%

Hair Cell Transduction Efficiency of Single- and Dual-AAV Serotypes in Adult Murine Cochleae

Omichi

Yoshimura

Shibata

et al. 2020

Molecular Therapy - Methods & Clinical Development

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“…The third improvement involves the use of recently reported novel AAVs that have a higher transfection rate. [11][12][13][14]16,69,70 We believe that with these improvements, USMB-mediated cochlear gene transfection via the RWM would become a useful tool that could be translated into human clinical applications.…”

Section: Limitations and Future Improvementsmentioning

confidence: 99%

“…3 This viral vector has been used in the gene therapy studies of auditory system in animal models [11][12][13][14][15] and human trials. 11,16 The inner ear is highly isolated from surrounding organs and tissues. This unique feature makes it an ideal organ for genetic manipulation, with a low risk of side effects.…”

Section: Introductionmentioning

confidence: 99%

Ultrasound‐microbubble cavitation facilitates adeno‐associated virus mediated cochlear gene transfection across the round‐window membrane

Zhang

Chen

Fan

et al. 2020

Bioengineering & Transla Med

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The round window of the cochlea provides an ideal route for delivering medicines and gene therapy reagents that can cross the round window membrane (RWM) into the inner ear. Recombinant adeno-associated viruses (rAAVs) have several advantages and are recommended as viral vectors for gene transfection. However, rAAVs cannot cross an intact RWM. Consequently, ultrasound-mediated microbubble (USMB) cavitation is potentially useful, because it can sonoporate the cell membranes, and increase their permeability to large molecules. The use of USMB cavitation for drug delivery across the RWM has been tested in a few animal studies but has not been used in the context of AAV-mediated gene transfection. The currently available large size of the ultrasound probe appears to be a limiting factor in the application of this method to the RWM. In this study, we used home-made ultrasound probe with a decreased diameter to 1.5 mm, which enabled the easy positioning of the probe close to the RWM. In guinea pigs, we used this probe to determine that (1) USMB cavitation caused limited damage to the outer surface layer or the RWM, (2) an eGFP-gene carrying rAAV could effectively pass the USMB-treated RWM and reliably transfect cochlear cells, and (3) the hearing function of the cochlea remained unchanged. Our results suggest that USMB cavitation of the RWM is a good method for rAAV-mediated cochlear gene transfection with clear potential for clinical translation. We additionally discuss several advantages of the small probe size. K E Y W O R D S cochlea, cochlear gene therapy, gene delivery, Guinea pigs, recombinant adeno-associated virus, round window membrane, ultrasonic microbubbles cavitation Zhen Zhang and Zhengnong Chen contributed equally to this study.

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“…The cause of hearing loss is extremely heterogeneous, and in many regions of the world, deaf people tend to marry with each other to form rather complex deaf families [33][34][35][36][37][38][39][40]. In one such family, we identified two separate genetic causes of hearing loss in distinct affected members, including the recessive c.235delC (p.L79Cfs * 3) mutation in GJB2 (III-2) and the dominant c.481C>T (p.R161C) mutation in SOX10 (II-1, III-1, and IV-1).…”

Section: Discussionmentioning

confidence: 99%

Targeted Next-Generation Sequencing Identifies Separate Causes of Hearing Loss in One Deaf Family and Variable Clinical Manifestations for the p.R161C Mutation inSOX10

Lin

2020

Neural Plasticity

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Hearing loss is the most common sensory deficit in humans. Identifying the genetic cause and genotype-phenotype correlation of hearing loss is sometimes challenging due to extensive clinical and genetic heterogeneity. In this study, we applied targeted next-generation sequencing (NGS) to resolve the genetic etiology of hearing loss in a Chinese Han family with multiple affected family members. Targeted sequencing of 415 deafness-related genes identified the heterozygous c.481C>T (p.R161C) mutation in SOX10 and the homozygous c.235delC (p.L79Cfs∗3) mutation in GJB2 as separate pathogenic mutations in distinct affected family members. The SOX10 c.481C>T (p.R161C) mutation has been previously reported in a Caucasian patient with Kallmann syndrome that features congenital hypogonadotropic hypogonadism with anosmia. In contrast, family members carrying the same p.R161C mutation in this study had variable Waardenburg syndrome-associated phenotypes (hearing loss and/or hair hypopigmentation) without olfactory or reproductive anomalies. Our results highlight the importance of applying comprehensive diagnostic approaches such as NGS in molecular diagnosis of hearing loss and show that the p.R161C mutation in SOX10 may be associated with a wide range of variable clinical manifestations.

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AAV-ie enables safe and efficient gene transfer to inner ear cells

Cited by 165 publications

References 38 publications

Hair Cell Transduction Efficiency of Single- and Dual-AAV Serotypes in Adult Murine Cochleae

Hair Cell Transduction Efficiency of Single- and Dual-AAV Serotypes in Adult Murine Cochleae

Ultrasound‐microbubble cavitation facilitates adeno‐associated virus mediated cochlear gene transfection across the round‐window membrane

Targeted Next-Generation Sequencing Identifies Separate Causes of Hearing Loss in One Deaf Family and Variable Clinical Manifestations for the p.R161C Mutation inSOX10

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