BIOLOGIC CHARACTERIZATION OF ENGINEERED HSV FOR THERAPY OF BRAIN TUMORS

我们的目标是开发和完善一种基于分子的策略， 使用基因工程HSV治疗恶性神经胶质瘤。为了 为了实现这一目标并确定人类HSV治疗剂，我们将 对每种基因进行广泛的生物学表征 在第一个项目中生产的工程HSV。我们将生产，滴定和 维持每种结构的参考库存，并将进行 对每一种（复制能力，基因表达， 细胞病变效应、神经毒力），以确定体内 功效测试被证明可接受的病毒将被提供给 第三个项目的研究人员和两个小组将定义反 基因工程HSV在体内的肿瘤效应 胶质瘤-SCID小鼠模型和侧腹肿瘤-裸鼠模型。我们还 将评估肿瘤细胞在被破坏后存活的逃逸机制， 基因工程HSV构建体， 作为溶瘤剂的递送和功效。我们还将评估 宿主对HSV治疗颅内胶质瘤免疫应答 使用C57 BL/6在局部（肿瘤中）和全身发展 小鼠作为同基因G1 -261恶性胶质瘤的宿主。For engineered 细胞因子基因已经插入的HSV，它们抑制 对HSV免疫应答或增强的抗肿瘤免疫应答将 被寻找。最后，对于那些选择的HSV，提供良好的抗肿瘤 对正常大脑损伤最小的影响，我们将进行评估 通过脑内给药进行人体临床试验的潜在安全性 在一种非常敏感的猿类灵长类动物Aotus trivirgatus身上进行测试。

Our objective is to develop and refine a molecular-based strategy for the treatment of malignant gliomas using genetically engineered HSV. In order to meet this objective and identify a human HSV therapeutic, we will undertake extensive biologic characterization of each genetically engineered HSV produced in the first Project. We will produce, titer and maintain reference stocks of each of these constructs and will conduct primary screening of each (replication competence, gene expression, cytopathic effect, neurovirulence) to determine suitability for in vivo efficacy testing. Viruses that prove to be acceptable will be provided to investigators in the third Project and both groups will define the anti- tumor effects of genetically engineered HSV in vivo using an intracerebral glioma-scid mouse model and in a flank tumor-nude mouse model. Further, we will assess escape mechanisms by which tumor cells survive destruction of genetically engineered HSV constructs in an effort to improve virus delivery and efficacy as an oncolytic agent. We will also evaluate the host immune response to HSV treatment of intracranial gliomas as it develops both locally (in the tumor) and systemically using the C57BL/6 mouse as a host for the syngeneic Gl-261 malignant glioma. For engineered HSV in which cytokine genes have been inserted, their ability to suppress immune responsiveness to HSV or enhance anti-tumor immune responses will be sought. Finally, for those selected HSV that offer excellent anti-tumor effects with minimal injury to normal brain, we will conduct and evaluate their potential safety for use in human clinical trials by intracerebral testing in a very sensitive simian primate, Aotus trivirgatus.