The application of clinical lithotripter shock waves on RNA nucleotide delivery to cells

Journal article

The delivery of genes into cells through the transfer of ribonucleic acids (RNAs) has been shown to cause a change in the level of target protein expression. RNA based transfection is conceptually more efficient than commonly delivered plasmid DNA because it does not require division or damage of the nuclear envelope thereby increasing the chances of the cell remaining viable. Shock waves (SWs) have been shown to induce cellular uptake by transiently altering the permeability of the plasma membrane, thereby overcoming a critical step in gene therapy. However, accompanied SW bioeffects include dose dependent irreversible cell injury and cytotoxicity. Here, the effect of SWs generated by a clinical lithotripter on the viability and permeabilisation of three different cell lines in vitro was investigated. Comparison of RNA stability before and after SW exposure showed no statistically significant difference. Optimal SW exposure parameters were identified to minimise cell death and maximise permeabilisation, and applied to enhanced green fluorescent protein (eGFP) messenger RNA (mRNA) or anti-eGFP small interfering RNA (siRNA) delivery. This resulted in eGFP mRNA expression levels increasing up to 52-fold in CT26 cells, whilst a 2-fold decrease in GFP expression was achieved following anti-eGFP siRNA delivery to MCF-7/GFP cells. These results demonstrate that SW parameters can be employed to achieve effective nucleotide delivery, laying the foundation for non-invasive and high safety RNA-based gene therapy.

Authors: Carlisle, R

• Publication status: Published
• Peer review status: Peer reviewed
• Publisher: Elsevier
• Journal: Ultrasound in Medicine and Biology
• Volume: 42
• Issue: 10
• Pages: 2478–2492
• Publication date: 2016-06-01
• Acceptance date: 2016-06-02
• DOI: 10.1016/j.ultrasmedbio.2016.06.001
• ISSN: 1879-291X
• Keywords: siRNA; shock waves; mRNA; drug delivery; ultrasound; high amplitude acoustic waves; gene therapy