CONSTRUCTION OF RECOMBINANT VIRUSES FOR DEVELOPMENT OF THERAPEUTIC APPLICATIONS

该计划项目拨款的总体目标是开发和测试 可以使用的基因工程单纯疱疹病毒(HSV) 安全有效地治疗人类恶性原发脑病 肿瘤。本项目的目标是构建一系列重组的 对恶性胶质瘤细胞具有高度选择性的HSV 这将产生直接的溶瘤作用或诱导溶瘤 直接去吧。我们建议提供至少5种新的基因工程 每年一次的HSV用于评估体外和相关的溶瘤活性 实验动物系统中的胶质瘤模型。为了实现这一目标 有效和高效地，我们将开发一种快速 基于宇宙的基因工程单纯疱疹病毒的构建。我们会 确定HSV基因组的最小包装容量和 将构建能够携带多种非病毒基因的载体。 最后，为了提供安全裕度并更好地控制时间 以及溶瘤作用的程度，我们将寻求创造条件 在这种情况下，特定基因的表达可以在体内通过 特定的推动者。它的目标是能够构建衰减的 适合脑内接种的遗传工程HSV 各种各样的实验动物系统(包括猿猴灵长类 比人类更敏感的物种)，并表现出 有可能区分正常细胞和恶性细胞。作为一种手段 为了放大溶瘤效应，我们将从基因上构建 携带外来基因的工程单纯疱疹病毒，其产物将 直接产生细胞毒性效应或将产生辐射 致敏作用或诱发抗肿瘤免疫相关炎症 回应。

The overall objective of this program project grant is to develop and test genetically engineered herpes simplex viruses (HSV) that can be used safely and effectively to treat humans with malignant primary brain tumors. The goal of this project is to construct a series of recombinant HSV which exhibit a high level of selectivity for malignant glioma cells and that will produce a direct oncolytic effect or will induce oncolysis directly. We propose to provide at least 5 novel genetically engineered HSV annually for assessment of oncolytic activity in vitro and in relevant glioma models in experimental animal systems. In order to achieve this effectively and efficiently, we will develop a method for rapid construction of genetically engineered HSV based on cosmids. We will determine the minimal the minimal packaging capacity of the HSV genome and will construct vectors capable of carrying multiple non-viral genes. Finally, to provide a margin of safety and to better control the timing and extent of the oncolytic effect, we will seek to create conditions under which expression of specific genes can be controlled in vivo using specific promoters. It is a goal to be able to construct attenuated genetically engineered HSV suitable for intracerebral inoculation into a wide variety of experimental animal systems (including simian primate species of greater susceptibility than humans) and that exhibit the potential to discriminate between normal and malignant cells. As a means of amplifying the oncolytic effect, we will construct genetically engineered HSV that carry foreign genes, the products of which will produce a cytotoxic effect directly or will act to induce a radiation sensitizing effect or to elicit an anti-tumor immune-related inflammatory response.