Optimization of Tumor Targeted HSV for Human Use

人用肿瘤靶向 HSV 的优化

• 批准号: 8299609
• 负责人: Bernard Roizman
• 金额: $ 35.79万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8299609
• 关键词: Affinity; Attenuated; Biochemistry; Biology; Cell Line; Cell surface; Cells; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Effectiveness; Engineering; Gene Expression; Gene Expression Profile; Generations; Genes; Genotype; Glucose; Glycoproteins; Growth; Herpesvirus 1; Human; Instruction; Interleukin-13; Ionizing radiation; Knowledge; Kringles; Ligands; Lysine; MEKs; Malignant Glioma; Methods; Modification; Molecular; Molecular Profiling; Normal Cell; PVRL1; Patients; Plasminogen; Preclinical Testing; Production; Program Research Project Grants; Progress Reports; Quality of life; Radiation; Resistance; Safety; Simplexvirus; Site; Specificity; System; Testing; Therapeutic; Time; Urokinase; Urokinase Plasminogen Activator Receptor; Validation; Virus; Virus Replication; activating transcription factor; base; cancer cell; design; design and construction; genetic regulatory protein; heparin proteoglycan; improved; mutant; novel; novel therapeutics; novel virus; pre-clinical; promoter; receptor; recombinant virus; tumor; virus development

Project 1 has had two major objectives. The first was to enhance effectiveness of candidate therapeutic viruses constructed by deletion of specific genes. These viruses are more effective in treatment that combines virus administration to the tumor and radiation. Nevertheless, the effectiveness of combined therapy is tumor genotype-dependent. In the past 5 years, we have collaborated with Project 2 to (a) identify the molecular basis for enhancement of attenuated viruses by ionizing radiation (IR) and (b) identify a cellular gene (MEK) whose product, when used properly in conjunction with IR, could overcome the restriction to attenuated virus replication imposed by tumor genotypes. The results of these studies are described in the Progress Report for Project 2. Our second objective was to construct viruses that can only infect cancer cells but not normal cells. In essence we ablated the ability of HSV-1 to attach to heparin sulfate proteoglycans and to enter cells by way of its natural protein receptors, HveA and nectin 1. Thus, the engineered virus R5141 infects cells solely via the IL13 a2 receptor while R5181 enters cells via the urokinase plasminogen activator receptor. Our findings have significantly contributed to understanding the biology and biochemistry of glycoprotein (g) D. Our new objectives are as follows: AIM 1 will develop methods for production of clinical grade targeted viruses. Since targeted viruses must be produced in noncancerous cells that stably express the novel receptor(s), we propose several ways in which we can produce clinical grade viruses. The objective of AIM 2 is to develop more effective viruses for therapy of GBM with AY34.5 mutant viruses. Therapeutic viruses currently in clinical trials extend survival time in a small fraction of treated patients in a tumor genotype dependent manner. We have constructed a virus in which the constitutively active MEK gene is driven by an IR inducible promoter (R2660). In preliminary studies R2660 plus IR blocked growth of a tumor resistant to virus or IR alone. The safety features of this and other mutant viruses will be studied. The objective of AIM 3 is to render the viruses targeting specific receptors on cell surfaces more effective. We have established proof of principle but do not consider the current generation optimal. We have identified specific shortcomings and ways to improve these viruses The objective of AIM 4 is based on the novel observation that a component of the amino-terminal domain of the urokinase plasminogen activator receptor interacts with the carboxyl-terminus of the gD ectodomain. The amino terminus of urokinase plasminogen activator contains a Kringle domain with affinity for lysines. Thus, we will determine whether the Kringle domain of human plasminogen can be used to target gD and by extension, target HSV-1 to its ligand, the glucose regulatory protein 78. RELEVANCE (See instructions): Project 1 is a component of a Program Project Grant designed to cure or at least effectively prolong quality of life of patients with malignant gliomas. At this point in time the proof of principle has been established at both basic and clinical levels. The task confronting Project 1 is to design, construct and test the next generation of therapeutic viruses characterized by enhanced therapeutic profile and to develop methods for their production in GMP facilities. Validation ofthe novel viruses will be done in collaboration with Projects 2 and 3 and, ultimately, with Project 4.

项目1有两个主要目标。第一个是提高候选治疗药物的有效性 通过删除特定基因而构建的病毒。这些病毒在治疗上更有效 结合了对肿瘤的病毒给药和放射治疗。然而，结合的有效性 治疗依赖于肿瘤基因型。在过去5年，我们与项目2合作， 通过电离辐射（IR）增强减毒病毒的分子基础和（B）鉴定a 细胞基因（MEK）的产物，当与IR结合使用时，可以克服 肿瘤基因型对减毒病毒复制的限制。这些研究的结果 项目2的进度报告中所述。我们的第二个目标是构建病毒， 感染癌细胞但不感染正常细胞。实质上，我们消除了HSV-1附着在肝素上的能力， 硫酸化蛋白聚糖并通过其天然蛋白受体HveA和nectin 1进入细胞。因此 工程化病毒R5141仅通过IL 13 α 2受体感染细胞，而R5181通过IL 13 α 2受体进入细胞。 尿激酶纤溶酶原激活物受体我们的研究结果大大有助于理解 糖蛋白生物学和生物化学（g）D.我们的新目标如下：AIM 1将开发 生产临床级靶向病毒的方法。由于目标病毒必须在 稳定表达新受体的非癌细胞，我们提出了几种方法， 产生临床级病毒。AIM 2的目标是开发更有效的病毒用于治疗 具有AY34.5突变病毒的GBM。目前在临床试验中的治疗性病毒延长了 一小部分治疗的患者以肿瘤基因型依赖的方式。我们制造了一种病毒 其中组成型活性MEK基因由IR诱导型启动子（R2660）驱动。初步 研究R2660加IR阻断了对病毒或单独IR具有抗性的肿瘤的生长。这款车的安全性能 和其他突变病毒将被研究。AIM 3的目的是使病毒靶向特异性 细胞表面的受体更有效。我们已经建立了原则证明，但不考虑 当前最佳代。我们已经确定了具体的缺点和改进这些病毒的方法 AIM 4的目的是基于新的观察结果， 尿激酶纤溶酶原激活物受体与gD胞外域的羧基末端相互作用。的 尿激酶纤溶酶原激活剂的氨基末端含有对赖氨酸具有亲和力的Kringle结构域。因此，在本发明中， 我们将确定人纤溶酶原的Kringle结构域是否可用于靶向gD， 在一个实施方案中，通过将HSV-1与其配体葡萄糖调节蛋白78靶向延伸，可以将HSV-1靶向其配体葡萄糖调节蛋白78。 相关性（参见说明）： 项目1是计划项目补助金的一部分，旨在治愈或至少有效地延长质量 恶性胶质瘤患者的生命周期。此时，原则证明已经确立， 基础和临床两个层面。项目1面临的任务是设计、建造和测试下一个项目。 产生以增强的治疗特性为特征的治疗性病毒，并开发用于 在GMP设施中生产。新病毒的验证将与项目2合作完成 第三个，最后是第四个项目。