Analysis of Gene Transfer Rate with Immobilized Retroviral Vectors

Abstract

Efficient delivery of transgenes into the cell nucleus by retroviral vectors in a static culture system is limited by the intrinsic features of incompetent retroviruses (i.e., thermodynamically unstable envelope proteins and low titers). Although several physicochemical approaches (e.g., adding polycationic polymer and applying magnetic force) have been reported to augment the retroviral gene transfer rate, none are suitable for scaling up to a setting for clinical use. The study of using acoustic fields with the form of standing waves has recently been reported to be a feasible way to enhance retroviral gene delivery efficiency in large-scale settings. The concept of using ultrasound standing-wave fields to increase retrovirus-mediated gene transfer is based on quickly established cell bands on acoustic nodal planes as nucleating sites to capture unstable colloidlike retroviruses. In this study, instead of having retroviral nanoparticles circulated between nodal planes, we proposed to immobilize retroviruses onto acoustic transparent films arranged in an acoustic chamber. Then, cells inoculated into the acoustic chamber can be driven by the primary radiation forces to the retrovirus-coated films that are constructed on the nodal planes. To obtain the optimal time of immobilizing retroviruses onto the acoustic transparent film prior to the inception of acoustic fields, we developed a retroviral diffusion-reaction model to describe such a static retroviral system. Analysis of viral transport model has its merit to guide experimental design for attaining high gene transfer efficiency.

Gene therapy for cardiovascular disease mediated by ultrasound and microbubbles

“…Viral systems derived from adeno-associated virus (AAV) [12], retrovirus [13], lentivirus [14] and adenovirus [15] are one of the successful gene delivery systems, which are used in the majority of current gene therapy researches and clinical trials due to their benefits of highly efficient delivery into cells with sustained expression. Recently, Prunier et al [16] showed that delivering adenovirus expressing sarcoplasmic reticulum Ca 2+ ATPase (SERCA2a) into coronary arteries could prevent ventricular arrhythmias in a ischemia - reperfusion model.…”