Arming Oncolytic HSV Vectors to Induce Anti-GBM Immune Responses in Syngeneic Mice

武装溶瘤 HSV 载体在同基因小鼠中诱导抗 GBM 免疫反应

基本信息

批准号：
10409654
负责人：
Joseph C Glorioso
金额：
$ 38.92万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-07 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10409654
关键词：
Adult Affect American Angiogenesis Inhibitors Animal Model Animals Antibodies Brain Neoplasms CD8-Positive T-Lymphocytes CTLA4 gene Cells Clinical Trials Cytotoxic T-Lymphocytes Dose Effectiveness Engineering Epidermal Growth Factor Receptor Evaluation Flow Cytometry Genes Genetically Engineered Mouse Glioblastoma HSV vector Herpesviridae Human Image Immune Immune Evasion Immune checkpoint inhibitor Immune response Immune system Immunization Immunocompetent Immunohistochemistry In Vitro Inbred BALB C Mice Infection Infiltration Injections Kinetics Lateral Lead Ligands Luciferases Lymphocyte Subtypings Lymphoid Cell Lytic MMP9 gene Macrophage Colony-Stimulating Factor Receptor Magnetic Resonance Imaging Malignant Neoplasms Malignant neoplasm of brain Matrix Metalloproteinases Mediating Metalloproteases MicroRNAs Modeling Mus Mutate NK Cell Activation Natural Immunity Natural Killer Cells Nature Nude Mice Oncolytic Oncolytic viruses Patients Performance Phenotype Population Prevalence Production Recombinants Recurrence Regulatory T-Lymphocyte Retroviridae Simplexvirus Sleeping Beauty T-Cell Depletion T-Lymphocyte T-Lymphocyte and Natural Killer Cell TIMP3 gene TP53 gene Testing Therapeutic Time Toxic effect Transforming Growth Factor beta Treatment Efficacy Treatment outcome Tropism Tumor Cell Line Tumor Immunity Tumor Tissue Tumor-associated macrophages Universities Up-Regulation Vegf Inhibitor Virion Virus Diseases Washington Xenograft procedure adaptive immunity antagonist anti-CTLA4 anti-PD-L1 anti-cancer anti-tumor immune response arm base brain cell brain tissue cell killing combinatorial cytokine design efficacy evaluation epidermal growth factor receptor VIII exhaustion experimental study immune activation immune checkpoint immunoregulation improved in vivo in vivo imaging macrophage mouse model neonate neoplastic cell neovascularization novel oncolysis oncolytic herpes simplex virus oncolytic vector overexpression programmed cell death ligand 1 reconstitution small hairpin RNA tumor tumor microenvironment tumor progression tumor-immune system interactions vector

项目摘要

Oncolytic HSV vectors (oHSV) offer considerable promise in the treatment of Glioblastoma Multiforme (GBM). In previous studies we designed a new base vector (KGN-4:T124) that is blocked for replication in normal brain tissue by brain-specific cellular miR-124 targeting of ICP4 expression. This base vector was further retargeted (KGNE-4:T124) to achieve selective infection of GBM cells expressing EGFR/EGFRvIII. Arming of this vector with a matrix metalloproteinase (KGNE-4:T124-MMP9) enhanced vector spread and oncolysis in xenogeneic animal models. However, virolysis alone is unlikely to achieve complete elimination of tumor cells that can cause recurrence. Effective elimination of all tumor cells will require the induction of innate and adaptive anti-tumor immunity. Unfortunately, GBM down-regulates the cellular machinery needed to sustain activation of immune cells including NK, macrophages and cytotoxic T cells. Therefore we propose to test the hypothesis that GBM treatment with our oHSV base vectors, KGN-4:T124 and KGNE-4:T124, can be enhanced through vector arming with immunomodulatory genes that inhibit tumor progression and induce innate and adaptive anti-tumor immune responses. To this end, we will initially exploit a tumor cell line (BAGL1) derived from a GBM tumor induced in BALB/c mice by Sleeping Beauty transposition of genes encoding luciferase, anti-p53 shRNA, N-RasV12 and human EGFRvIII. These experiments will be extended to a genetically-induced GBM model developed by Dr. Eric Holland that avoids tumor cell injection. Our aims will seek to counteract three aspects of immune evasion in GBM: (i) ADAM17-mediated shedding of NK cell activators and reduced NK function, (ii) expression of the immune checkpoint molecules that lead to NK and T cell exhaustion, and (iii) the presence of M2 macrophages that contribute to an immunosuppressive tumor microenvironment (TME). In Aim 1, we will test the hypothesis that our unarmed base vectors, collectively referred to as KGN(E)-4:T124, will provide an effective GBM treatment in immune competent mice. We will determine the impact of vector dosing on intratumoral vector distribution, TME composition and animal survival, and assess the value of EGFR targeting. We will also examine the effect of prior HSV immunization on treatment outcomes. In Aim 2, we will test the hypothesis that KGN(E)- 4:T124 derivatives armed with TIMP-3 (ADAM17 inhibitor/VEGF-R2 antagonist) will improve therapy. We will evaluate the impact of TIMP-3 expression on NK cell activation, tumor spread and animal survival. In Aim 3, we will test the hypothesis that vectors armed with single chain antibody (scFv) checkpoint inhibitors (anti-PD-L1 or anti-CTLA4), alone or in combination with depletion of the immunosuppressive (M2) TAM population using BLZ945-mediated inhibition of colony-stimulating factor-1 receptor (CSF-1R), will produce effective anti-tumor immunity, particularly in HSV immune mice. Anti-tumor immunity will be established using a bi-lateral tumor model and confirmed by selective antibody-mediated T cell depletion.

溶瘤 HSV 载体 (oHSV) 在治疗多形性胶质母细胞瘤 (GBM) 方面具有巨大的前景。在之前的研究中，我们设计了一种新的基本载体（KGN-4:T124），该载体在正常大脑中的复制被阻断通过脑特异性细胞 miR-124 靶向 ICP4 表达来控制组织。该基本向量被进一步重定向 (KGNE-4:T124)以实现表达EGFR/EGFRvIII的GBM细胞的选择性感染。该向量的武装使用基质金属蛋白酶 (KGNE-4:T124-MMP9) 增强载体在异种中的扩散和溶瘤作用动物模型。然而，单独的病毒溶解不太可能实现完全消除肿瘤细胞，从而导致复发。有效消除所有肿瘤细胞需要诱导先天性和适应性抗肿瘤细胞免疫。不幸的是，GBM 下调了维持免疫激活所需的细胞机制。细胞包括NK细胞、巨噬细胞和细胞毒性T细胞。因此，我们建议检验 GBM 的假设我们的 oHSV 基本载体 KGN-4:T124 和 KGNE-4:T124 的治疗可以通过载体武装来增强具有抑制肿瘤进展并诱导先天性和适应性抗肿瘤免疫的免疫调节基因回应。为此，我们将首先开发源自 GBM 肿瘤的肿瘤细胞系 (BAGL1) BALB/c 小鼠通过睡美人转座编码荧光素酶、抗 p53 shRNA、N-RasV12 和人 EGFRvIII。这些实验将扩展到由 Dr. 开发的遗传诱导的 GBM 模型。埃里克·霍兰德（Eric Holland）避免注射肿瘤细胞。我们的目标将寻求抵消免疫逃避的三个方面 GBM 中：(i) ADAM17 介导的 NK 细胞激活剂脱落和 NK 功能降低，(ii) 导致 NK 和 T 细胞耗竭的免疫检查点分子，以及 (iii) M2 巨噬细胞的存在有助于形成免疫抑制肿瘤微环境（TME）。在目标 1 中，我们将检验假设我们的非武装基础向量（统称为 KGN(E)-4:T124）将提供有效的 GBM 对免疫能力强的小鼠进行治疗。我们将确定载体剂量对瘤内载体的影响分布、TME 组成和动物存活，并评估 EGFR 靶向的价值。我们还将检查先前的 HSV 免疫对治疗结果的影响。在目标 2 中，我们将检验假设 KGN(E)- 4：配备 TIMP-3（ADAM17 抑制剂/VEGF-R2 拮抗剂）的 T124 衍生物将改善治疗。我们将评估 TIMP-3 表达对 NK 细胞活化、肿瘤扩散和动物存活的影响。在目标 3 中，我们将检验载有单链抗体 (scFv) 检查点抑制剂（抗 PD-L1 或抗 CTLA4)，单独使用或与免疫抑制 (M2) TAM 群体的消除组合使用 BLZ945介导的集落刺激因子1受体（CSF-1R）抑制，将产生有效的抗肿瘤作用免疫，特别是HSV免疫小鼠。利用双侧肿瘤建立抗肿瘤免疫力模型并通过选择性抗体介导的 T 细胞耗竭得到证实。