Project 4: Awakening immune responses to GBM by enhancing immune cell trafficking and activation with oHSV armed with Cetuximab-CCL5 and anti-CD47 antibody payloads.

项目 4：通过配备西妥昔单抗-CCL5 和抗 CD47 抗体有效负载的 oHSV 增强免疫细胞运输和激活，唤醒对 GBM 的免疫反应。

• 批准号: 10712283
• 负责人: MICHAEL A CALIGIURI
• 金额: $ 35.5万
• 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 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10712283
• 关键词: Adenosine; Adoptive Cell Transfers; Anti-CD47; Antibodies; Binding; Biodistribution; Biological Markers; Brain; CCR1 gene; CD47 gene; Cells; Cetuximab; Circulation; Cities; Clinical; Clinical Trials; Combined Modality Therapy; Data; Development; Dose; Eating; Engineering; Epidermal Growth Factor Receptor; Fc Receptor; Future; Glioblastoma; Goals; Good Manufacturing Process; Growth Factor; Herpes Simplex Infections; Herpesvirus 1; Human; IgG1; Immune; Immune response; Immunity; Immunocompetent; Immunologic Stimulation; Immunosuppression; Immunotherapy; In Vitro; Inflammatory; Investigational Drugs; Investments; Length; Link; Macrophage; Malignant - descriptor; Malignant Glioma; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Modeling; Monitor; Monoclonal Antibodies; Mus; NK Cell Activation; Natural Killer Cells; Normal tissue morphology; Oncolytic; Oncolytic viruses; Outcome; PD-1/PD-L1; PTPNS1 gene; Patients; Production; Publications; Publishing; RANTES; Research; Safety; Sampling; Schedule; Signal Transduction; Solid Neoplasm; Surface; System; T-Lymphocyte; Testing; Tissues; Toxic effect; Tumor-infiltrating immune cells; Viral; Virus; Virus Replication; anti-PD-1; anti-tumor immune response; antibody-dependent cell cytotoxicity; antibody-dependent cellular phagocytosis; arm; chemokine; chimeric antigen receptor T cells; cytokine; design; efficacy study; immune cell infiltrate; immune checkpoint blockade; immunoregulation; improved; in vivo; innovation; manufacture; migration; mouse model; neoplastic cell; novel; oncolysis; oncolytic virotherapy; patient prognosis; peripheral blood; pharmacokinetics and pharmacodynamics; pre-clinical research; preclinical efficacy; preclinical safety; preclinical study; prevent; receptor; recruit; response; scaffold; synergism; trafficking; treatment optimization; tumor; tumor microenvironment; tumor progression

PROJECT SUMMARY – PROJECT 4 The ultimate goal of this proposal is to develop a novel, effective, and safe oncolytic herpes simplex viral (oHSV)- based immunotherapy for the treatment of glioblastoma (GBM), a highly fatal and the most common malignant brain tumor. oHSV treatment of GBM relies on cancer-specific replication of the virus leading to tumor destruction with minimal toxicity to adjacent non-neoplastic tissue. Its safety in patients has been proven, yet efficacy remains to be improved. Project 4 will focus on enhancing anti-tumor immune responses by arming oHSVs with powerful immune-modulation payloads by testing two newly generated novel oHSVs, termed OV-Cmab-CCL5 and OV- αCD47-IgG1, designed to induce immune infiltration and block a checkpoint, respectively, as monotherapies in addition to their combination. The two oHSVs are expected to synergize with each other to maximize immune responses in the GBM microenvironment by targeting both immune stimulation and immune suppression to create an overall pro-inflammatory tumor microenvironment, which has been demonstrated to positively correlate with patient survival in the rQNestin34.5 clinical trial of Project 2. We generated OV-Cmab-CCL5, an oHSV expressing a secretable single-chain variable fragment of the epidermal growth factor receptor (anti-EGFR) IgG1 antibody cetuximab linked to CCL5 by an Fc knob-into-hole system that produces heterodimers and prevents homodimers. To target a checkpoint on innate immune cells, we also engineered oHSV to express a full-length, soluble anti-CD47 antibody with a human IgG1 scaffold (so-called OV-αCD47-G1), for locoregional control of GBM. The antibody αCD47-IgG1 secreted by virus-infected GBM cells blocks the CD47 “don't eat me” signal and exerts additional functions of Fc receptor-mediated antibody-dependent cellular phagocytosis by macrophages and antibody-dependent cellular cytotoxicity by NK cells. Our data show that in GBM mouse models, both oHSVs reduced tumor size and prolonged survival, owing to enhanced anti-tumor immune responses. We hypothesize that OV-Cmab-CCL5 is a safe and effective agent that can improve GBM therapy with multiple mechanisms of action, and the combination therapy of OV-Cmab-CCL5 and OV-αCD47-IgG1 will have better efficacy than the respective monotherapies. We have manufactured Good Manufacturing Practice (GMP)-grade viruses at our in-house GMP facility and now propose to conduct Investigational New Drug (IND) enabling studies so that our innovative oHSVs will be ready for testing in future clinical trials. Project 4 has three Specific Aims: (1) dissect systemic and regional immune responses, identify a marker(s) in the peripheral blood of mouse models or clinical samples from Project 2 that correlates with anti-tumor activity, and improve the efficacy of rQnestin34.5.v2 in GBM mouse models after OV-Cmab-CCL5 treatment; (2) perform IND-enabling in vivo efficacy and toxicity studies using GMP-grade OV-Cmab-CCL5; and (3) determine the preclinical efficacy and safety of OV-Cmab-CCL5 combined with OV-αCD47-IgG1. We are confident that our approaches will provide a novel, effective, and safe oncolytic virotherapy against GBM.

项目概要-项目4 该提案的最终目标是开发一种新型、有效和安全的溶瘤性单纯疱疹病毒（oHSV）， 用于治疗胶质母细胞瘤（GBM）的基于免疫疗法，胶质母细胞瘤是一种高度致命且最常见的恶性肿瘤， 脑瘤GBM的oHSV治疗依赖于导致肿瘤破坏的病毒的癌症特异性复制 对邻近非肿瘤组织的毒性最小。它在患者中的安全性已被证明，但疗效仍然存在 有待改进。项目4将重点关注通过用强大的抗肿瘤药物武装oHSV来增强抗肿瘤免疫应答。 免疫调节有效载荷，通过测试两种新产生的新型oHSV，称为OV-Cmab-CCL 5和OV- α CD 47-IgG 1，分别用于诱导免疫浸润和阻断检查点，作为单药治疗， 除了他们的组合。这两种oHSV有望相互协同，以最大限度地提高免疫力。 通过靶向免疫刺激和免疫抑制， 创造了一个整体的促炎性肿瘤微环境，这已被证明是正相关的 与项目2的rQNestin34.5临床试验中的患者生存率相关。我们产生了OV-CmAb-CCL 5，一种oHSV 表达表皮生长因子受体（抗EGFR）IgG 1的分泌型单链可变片段 抗体西妥昔单抗通过Fc杵入臼系统与CCL 5连接，该系统产生异二聚体并阻止 同源二聚体。为了靶向先天免疫细胞上的检查点，我们还设计了oHSV以表达全长， 可溶性抗CD 47抗体与人IgG 1支架（所谓的OV-α CD 47-G1），用于局部控制 GBM。病毒感染GBM细胞分泌的抗体α CD 47-IgG 1阻断CD 47“不要吃我”信号 并通过以下方式发挥Fc受体介导的抗体依赖性细胞吞噬作用的额外功能： 巨噬细胞和NK细胞的抗体依赖性细胞毒性。我们的数据显示，在GBM小鼠中， 在模型中，由于增强的抗肿瘤免疫，两种oHSV都减小了肿瘤大小并延长了生存期， 应答我们假设OV-CmAb-CCL 5是一种安全有效的药物，可以改善GBM治疗 具有多种作用机制，OV-CmAb-CCL 5和OV-α CD 47-IgG 1的联合治疗将 具有比各自的单一疗法更好的功效。我们已经制定了良好的生产规范 （GMP）级病毒在我们的内部GMP设施，现在建议进行研究性新药（IND） 使研究，使我们的创新oHSV将准备在未来的临床试验测试。项目4有三个 具体目的：（1）解剖全身和局部免疫反应，确定外周血中的标记物 与抗肿瘤活性相关的小鼠模型或来自项目2的临床样品， 在OV-Cmab-CCL 5处理后，rQnestin34.5.v2在GBM小鼠模型中的功效;（2）在GBM小鼠模型中进行IND使能， 使用GMP级OV-CmAb-CCL 5的体内功效和毒性研究;和（3）确定临床前功效 OV-Cmab-CCL 5联合OV-α CD 47-IgG 1的安全性。我们相信，我们的方法将 提供了一种新的、有效的和安全的针对GBM的溶瘤病毒疗法。