Optimization of Tumor Targeted HSV for Human Use

人用肿瘤靶向 HSV 的优化

• 批准号: 8080211
• 负责人: Bernard Roizman
• 金额: $ 37万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8080211
• 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; 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。首先是提高候选治疗的有效性 通过缺失特定基因构建的病毒。这些病毒在治疗方面更有效 将病毒施用结合到肿瘤和辐射。然而，联合的有效性 治疗是肿瘤基因型依赖性的。在过去的5年中，我们与项目2至（a）确定的项目2合作 通过电离辐射（IR）和（b）识别A的分子基础，以增强衰减病毒 蜂窝基因（MEK），当与IR结合使用时，其产物可以克服 限制肿瘤基因型施加的减毒病毒复制。这些研究的结果是 在项目2的进度报告中描述的。我们的第二个目标是构建只能 感染癌细胞，而不是正常细胞。本质上，我们消除了HSV-1附着在肝素上的能力 硫酸盐蛋白聚糖并通过其天然蛋白质受体Hvea和Nectin 1进入细胞。因此， 设计的病毒R5141仅通过IL13 A2受体感染细胞，而R5181通过 尿激酶纤溶酶原活化剂受体。我们的发现极大地有助于理解 糖蛋白的生物学和生物化学（G）D。我们的新目标如下：AIM 1将发展 生产临床等级靶向病毒的方法。由于必须在 稳定表达新型受体的非癌细胞，我们提出了几种方法 产生临床级病毒。目标2的目的是开发更有效的病毒来治疗 具有AY34.5突变病毒的GBM。目前在临床试验中的治疗病毒在A中延长了生存时间 以肿瘤基因型依赖性方式治疗的患者的一小部分。我们已经在 组成型活性MEK基因是由IR诱导启动子（R2660）驱动的。在初步 研究R2660加IR阻断了对病毒或IR抗性肿瘤的生长。安全特征 将研究其他突变病毒。目标3的目的是渲染针对特定的病毒 细胞表面上的受体更有效。我们已经建立了原则证明，但不考虑 当前一代最佳。我们已经确定了具体的缺点和改善这些病毒的方法 目标4的目的是基于新的观察结果，即氨基末端结构域的组成 尿激酶纤溶酶原活化剂受体与GD外生域的羧基末端相互作用。这 尿激酶纤溶酶原激活剂的氨基末端包含一个具有对赖氨酸亲和力的kringle结构域。因此， 我们将确定是否可以使用人纤溶酶原的kringle结构域来靶向GD和 延伸至其配体的靶标HSV-1，葡萄糖调节蛋白78。 相关性（请参阅说明）： 项目1是计划项目赠款的组成部分，旨在治愈或至少有效延长质量 恶性神经胶质瘤患者的寿命。此时，原则证明已在 基本和临床水平。面对项目1的任务是设计，构建和测试下一个 以增强的治疗特征和开发方法的特征的治疗病毒生成 他们在GMP设施中的生产。新型病毒的验证将与项目2合作完成 和3，最终与项目4。