Christ Moonen

Spatial and temporal control of transgene expression in vivo using a heat‐sensitive promoter and MRI‐guided focused ultrasound

Among the techniques used to induce and control gene expression, a non‐invasive, physical approach based on local heat in combination with a heat‐sensitive promoter represents a promising alternative but requires accurate temperature control in vivo. MRI‐guided focused ultrasound (MRI‐FUS) with real‐time feedback control allows automatic execution of a predefined temperature‐time trajectory. The purpose of this study was to demonstrate temporal and spatial control of transgene expression based on a well‐defined local hyperthermia generated by MRI‐FUS.

Image-guided control of transgene expression based on local hyperthermia.

Spatial and temporal control of transgene expression is one of the major prerequisites of efficient gene therapy. Recently, a noninvasive, physical approach has been presented based on local heat in combination with a heat-sensitive promoter. This strategy requires tight temperature control in vivo. Here, we use MRI-guided focused ultrasound (MRI-FUS) with real-time feedback control on a whole-body clinical MRI system for a completely automatic execution of a predefined temperature-time trajectory in the focal point. Feasibility studies on expression control were carried out on subcutaneously implanted rat tumors. A stable modified C6 glioma cell line was used carrying a fused gene coding for thymidine kinase (TK) and green fluorescent protein (GFP) under control of the human heat-shock protein 70 (HSP70) promoter. In vitro studies showed strong induction of the TK-GFP gene expression upon heat shock under various conditions and localization of the protein product in the nucleus. In vivo tumors were subjected to a 3-min temperature elevation using MRI-FUS with a constant temperature, and were analysed 24 hr after the heat shock with respect to GFP fluorescence. Preliminary results showed strong local induction in regions heated above 40°C, and a good correspondence between temperature maps at the end of the heating period and elevated expression of TK-GFP.