Quantitative Analysis of Retroviral Gene Delivery

该项目是一个未经请求的提交。 本项目的目标是开发策略，以提高逆转录病毒介导的基因转移的效率和效用。 逆转录病毒是目前基因治疗临床试验中广泛使用的载体系统，这导致了逆转录病毒载体技术的迅速发展。 尽管如此，逆转录病毒基因转移效率仍然令人不安地低，阻碍了逆转录病毒基因治疗作为一种有效的临床工具的出现。 已经发现，带电分子极大地影响基因转移过程的效率。 具体而言，带正电荷的化合物已被证明可以增强基因转移，而带负电荷的化合物则抑制该过程。 该项目旨在实现以下目标：（1）详细研究带电化合物增强和抑制逆转录病毒基因转移过程的机制，（2）开发用于分析带电化合物对病毒转运和转导的作用的数学框架，以及（3）为了比较使用优化的离体和体内基因治疗方案在动物模型中的基因转移效率，载体和靶细胞。为了实现这些目标，研究人员将利用一套互补的酶和免疫组织化学测定，分子成像技术，和细胞荧光分析，以定量逆转录病毒基因转移过程的各个步骤。 工程分析所得的生物分子运输和动力学反应数据将被用来促进逆转录病毒运输的综合模型的建设。

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.