Ultrasound-microbubble cavitation facilitates adeno-associated virus mediated cochlear gene transfection across the round-window membrane.
Zhen ZhangZhengnong ChenLiqiang FanThomas LandryJeremy BrownZhiping YuShankai YinJian WangPublished in: Bioengineering & translational medicine (2020)
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.
Keyphrases
- gene therapy
- magnetic resonance imaging
- copy number
- genome wide
- living cells
- quantum dots
- drug delivery
- genome wide identification
- sars cov
- dna methylation
- hearing loss
- induced apoptosis
- gene expression
- oxidative stress
- mesenchymal stem cells
- bone marrow
- fluorescent probe
- contrast enhanced ultrasound
- single molecule
- transcription factor
- cancer therapy
- newly diagnosed
- optical coherence tomography
- human health