Project 1: Arming Oncolytic HSV Vectors to Improve Virolysis in Syngeneic Mouse Models of GBM

项目 1：武装溶瘤 HSV 载体以改善 GBM 同基因小鼠模型中的病毒溶解

• 批准号: 10019362
• 负责人: Joseph C Glorioso
• 金额: $ 33.92万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-07 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019362
• 关键词: Aftercare; American; Animal Model; Animals; Antigens; Antiviral Agents; Binding; Binding Sites; Blocking Antibodies; Brain; Brain Neoplasms; Cell-Mediated Cytolysis; Cells; Clinical Trials; Collaborations; Cytomegalovirus; Data; Development; Dose; Effectiveness; Engineering; Environment; Epidermal Growth Factor Receptor; Evaluation; FCGR3B gene; Flow Cytometry; Funding; Future; Genes; Genetic; Glioblastoma; Glioma; Glycoproteins; Grant; HSV vector; Herpesviridae; Herpesvirus 1; Human; Immune; Immunize; Immunocompetent; Immunoglobulin G; Immunohistochemistry; In Vitro; Infection; Injections; Laboratories; Ligands; Lytic; Lytic Virus; MMP9 gene; Macrophage Activation; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Matrix Metalloproteinases; Mediating; Mediator of activation protein; Membrane Proteins; Modeling; Mus; NK Cell Activation; Natural Killer Cells; Oncolytic; Oncolytic viruses; Patient Recruitments; Population; Proteins; Recombinants; Role; Safety; Series; Signal Transduction; Simplexvirus; Site; Surface; System; TIMP3 gene; Testing; Time; Transgenes; Treatment Effectiveness; Treatment Efficacy; Tumor Immunity; Vertebral column; Viral; Virus; Virus Diseases; Virus Replication; Washington; anti-cancer; antibody-dependent cell cytotoxicity; arm; base; comparative efficacy; cytotoxicity; design; epidermal growth factor receptor VIII; experimental study; expression vector; glioma cell line; immunological status; implantation; improved; in vivo; in vivo evaluation; insight; macrophage; mouse model; neoplastic cell; notch protein; oncolysis; oncolytic herpes simplex virus; oncolytic vector; oncolytic virotherapy; phase I trial; prevent; programs; receptor; response; retroviral transduction; standard of care; treatment comparison; tumor; tumor microenvironment; tumor-immune system interactions; vector

PROJECT SUMMARY – PROJECT 1 Oncolytic herpes simplex virus (oHSV) vectors offer considerable promise in the treatment of Glioblastoma Multiforme (GBM). In the current funding period we designed a new series of HSV vectors capable of safe but efficient tumor destruction through (i) full vector retargeting to prevalent human GBM receptors (EGFR/EGFRvIII) (vector KGNE), (ii) cellular miR-124 blockade of vector replication in normal brain (KGN-4:T124: our base vector), or (iii) a combination of these features (KGNE-4:T124). Further arming of the combination vector with a matrix metalloproteinase (KGNE-4:T124-MMP9) enhanced vector spread and oncolysis in animal models. However, vector replication in tumors can still be limited by Natural Killer (NK) cells that normally protect the host from HSV infection. Projects 2 and 4 have recently demonstrated that interference with NK cell activation and killing of virus-infected cells enhances GBM treatment by allowing more robust and broader oncolytic virus spread. We propose to validate and extend these findings by comparing the impact of vector transgene-mediated NK blockade on the oncolytic activities of the two base vectors of this PPG, our KGN-4:T124 (strain KOS) and vector rQNestin34.5 (strain F) from Project 2, in three distinct syngeneic mouse models of GBM. Because these base vectors have different genetic backgrounds and are controlled by different safety mechanisms, their comparison in these experiments will be important for choosing the most effective backbone and arming genes for future trials. We will exploit two HSV-permissive mouse glioma cell lines, CT2A and GL261N4, to generate brain tumors in immunocompetent mice by tumor cell injection. In addition, we have an established collaboration with Eric Holland (U. of Washington) to verify key results in his genetically induced mouse models of GBM based on the RCAS/tv-a system. In Aim 1, we will establish base-line tumor treatment efficacies for KGN-4:T124 versus rQNestin34.5. We will perform tumor-treatment dose-response analyses, and characterize and compare the effects of the two vectors on the composition of the tumor microenvironment (TME). In Aim 2, we will take advantage of these base-line data to evaluate the effect of NK cell antagonism on tumor therapy. In collaboration with Projects 2 and 4, we will determine the effects of vector arming with genes encoding soluble forms of HCMV UL141 and/or PRV glycoprotein D that reduce the expression of glioma cell ligands recognized by NK cells. In Aim 3, we will express the Fcγ-binding ectodomains of two other HCMV-derived proteins, gp34 and gp68, that are known to inhibit NK cell activation. In collaborative experiments with Project 4, we will test the hypothesis that sgp34 will inhibit classical ADCC (antibody-dependent cellular cytotoxicity), enhance vector replication and tumor virolysis while sgp34 and/or sgp68 will interfere with Fc bridging cellular cytotoxicity (FcBCC). We will also provide TIMP-3 expression vectors to Project 3 to inhibit Notch signaling among macrophages and thereby reduce macrophage recognition of infected tumor cells and prolong oHSV persistence.

项目摘要 - 项目1 溶瘤疱疹单纯疱疹病毒（OHSV）载体在治疗胶质母细胞瘤方面有很大的希望 多形（GBM）。在当前的资金期间，我们设计了一个新系列的HSV向量，能够安全但 通过（i）全面载体重新定位到普遍的人类GBM受体（EGFR/EGFRVIII）的有效肿瘤破坏 （载体kgne），（ii）正常脑中矢量复制的细胞miR-124封锁（KGN-4：T124：我们的基本载体）， 或（iii）这些特征的组合（kgne-4：t124）。用矩阵进一步武装组合矢量 金属蛋白酶（KGNE-4：T124-MMP9）在动物模型中增强了载体扩散和肿瘤。然而， 肿瘤中的矢量复制仍然可以受到通常保护宿主免受HSV的天然杀手（NK）细胞的限制 感染。项目2和4最近证明了对NK细胞激活和杀戮的干扰 感染病毒的细胞通过允许更健壮和更广泛的溶瘤病毒扩散来增强GBM治疗。我们 通过比较向量变换介导的NK的影响来验证和扩展这些发现的建议 该PPG的两个基本载体的溶瘤活性，我们的KGN-4：T124（菌株KOS）和载体 RQNESTIN34.5（菌株F）来自项目2，在三种不同的GBM的合成小鼠模型中。因为这些基础 向量具有不同的遗传背景，并由不同的安全机制控制，它们的比较 在这些实验中，对于为将来选择最有效的骨干和武装基因将很重要 试验。我们将利用两种HSV验证小鼠神经胶质瘤细胞系CT2A和GL261N4产生脑肿瘤 通过肿瘤细胞注射在免疫能力的小鼠中。此外，我们与Eric建立了合作 根据 RCAS/TV-A系统。在AIM 1中，我们将建立KGN-4的碱基肿瘤治疗效率：T124与 RQNESTIN34.5。我们将执行肿瘤治疗剂量反应分析，并表征和比较 两个向量对肿瘤微环境（TME）组成的影响。在AIM 2中，我们将采取 这些基线数据的优势以评估NK细胞拮抗对肿瘤疗法的影响。合作 使用项目2和4，我们将用编码固体形式的HCMV的基因来确定向量武装的影响 UL141和/或PRV糖蛋白D减少NK细胞识别的神经胶质瘤细胞配体的表达。在 AIM 3，我们将表达另外两种HCMV衍生蛋白GP34和GP68的Fcγ结合的外生委员会， 已知可以抑制NK细胞活化。在与项目4的协作实验中，我们将检验假设 SGP34将抑制经典ADCC（抗体依赖性细胞毒性），增强矢量复制和 肿瘤病毒分析而SGP34和/或SGP68会干扰FC桥接细胞细胞毒性（FCBCC）。我们也会 向项目3提供TIMP-3表达向量，以抑制巨噬细胞之间的缺口信号传导，从而抑制 减少对感染肿瘤细胞的巨噬细胞识别并延长OHSV持久性。