Quantitative Analysis of Retroviral Gene Delivery

This project is an unsolicited submission. The goal of this project is to develop strategies to enhance the efficiency and utility of retrovirus-mediated gene transfer. Retroviruses are a widely utilized vector system in current gene therapy clinical trials, which has resulted in the rapid advancement of retroviral vector technology. Despite this, retroviral gene transfer efficiency has remained disappointingly low, hampering the emergence of retroviral gene therapy as an effective clinical tool. It has been found that charged molecules greatly influence the efficiency of the gene transfer process. Specifically, positively charged compounds have been shown to enhance gene transfer, whereas negatively charged compounds inhibit the process. This project seeks to accomplish the following: (1) to investigate in detail the mechanism of charged compound enhancement and inhibition of the retroviral gene transfer process, (2) to develop a mathematical framework for analyzing the effect of charged compounds on virus transport and transduction, and (3) to compare gene transfer efficiency in an animal model using ex vivo and in vivo gene therapy protocols optimized to enhance the electrostatic interaction between the retrovirus vector and target cell. To achieve these goals, the investigators will utilize a complementary set of enzymatic and immunohistochemical assays, molecular imaging techniques, and cytofluorometric analyses to quantitate various steps of the retroviral gene transfer process. Engineering analysis of the resultant biomolecular transport and kinetic reaction data will be used to facilitate the construction of an integrative model of retrovirus transport.