AAV2.7m8 is a powerful viral vector for inner ear gene therapy

Isgrig, Kevin; McDougald, Devin S.; Zhu, Jianliang; Wang, Hong Jun; Bennett, Jean; Chien, Wade W.

doi:10.1038/s41467-018-08243-1

Cited by 117 publications

(87 citation statements)

References 28 publications

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“…However, the precise time frame between the onset of Emx2 expression driven by Gfi1-Cre and reversal of centriole positions remains unclear. To further investigate the time required by Emx2 to alter hair bundle orientation, we ectopically expressed Emx2 in utricular explants using AAV2.7m8 adeno-associated virus, which has been shown to infect cochlear HCs efficiently (Isgrig et al, 2019). We infected CAG:GFP-Centrin2 utricular explants at E13.5 with an AAV2.7m8 adeno-associated viral vector, AAV2.7m8-CAG-Emx2-P2A-tdTomato (AAV-Emx2-tdT), in which both Emx2 and tdTomato transcripts were driven under the universal CAG promoter.…”

Section: Infections With Aav-emx2 Reverse Hair Bundle Orientationmentioning

confidence: 99%

Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2

Tona

2020

eLife

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The asymmetric hair bundle on top of hair cells (HCs), comprises a kinocilium and stereocilia staircase, dictates HC's directional sensitivity. The mother centriole (MC) forms the base of the kinocilium, where stereocilia are subsequently built next to it. Previously we showed that transcription factor Emx2 reverses hair bundle orientation and its expression in the mouse vestibular utricle is restricted, resulting in two regions of opposite bundle orientation (Jiang et al, 2017). Here, we investigated establishment of opposite bundle orientation in embryonic utricles by live-imaging GFP-labeled centrioles in HCs. The daughter centriole invariably migrated ahead of the MC from the center to their respective peripheral locations in HCs. Comparing HCs between utricular regions, centriole trajectories were similar but they migrated towards opposite directions, suggesting that Emx2 pre-patterned HCs prior to centriole migration. Ectopic Emx2, however, reversed centriole trajectory within hours during a critical time-window when centriole trajectory was responsive to Emx2.

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Section: Infections With Aav-emx2 Reverse Hair Bundle Orientationmentioning

confidence: 99%

Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2

Tona

2020

eLife

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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%

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 adeno-associated virus (AAV) is widely used in gene therapy due to its high infection efficiency, low pathogenicity and toxicity, sustained expression of the carried genes, as well as its simple, cheap, and fast production [21][22][23][24]. Several studies have achieved good results by AAV vector-mediated gene therapy using animal models with different mutated genes in several types of cells in cochlea [2,21,[25][26][27][28][29][30].…”

Section: A Brief Gene Therapy History In Auditory Diseasementioning

confidence: 99%

Recent development of AAV-based gene therapies for inner ear disorders

et al. 2020

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Gene therapy for auditory diseases is gradually maturing. Recent progress in gene therapy treatments for genetic and acquired hearing loss has demonstrated the feasibility in animal models. However, a number of hurdles, such as lack of safe viral vector with high efficiency and specificity, robust deafness large animal models, translating animal studies to clinic etc., still remain to be solved. It is necessary to overcome these challenges in order to effectively recover auditory function in human patients. Here, we review the progress made in our group, especially our efforts to make more effective and cell typespecific viral vectors for targeting cochlea cells.

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AAV2.7m8 is a powerful viral vector for inner ear gene therapy

Cited by 117 publications

References 28 publications

Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2

Live imaging of hair bundle polarity acquisition demonstrates a critical timeline for transcription factor Emx2

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

Recent development of AAV-based gene therapies for inner ear disorders

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