Project 4:Targeting M2-like Macrophages and MDSC with Myelolytic-Virotherapy

项目 4：利用溶髓病毒疗法靶向 M2 样巨噬细胞和 MDSC

• 批准号: 10885263
• 负责人: TIMOTHY P CRIPE
• 金额: $ 36.69万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10885263
• 关键词: Adoptive Transfer; CXCL10 gene; Cancer Etiology; Cancer Vaccines; Cell Therapy; Cells; Cellular immunotherapy; Chemotactic Factors; Childhood; Clinical; Combined Modality Therapy; Data; Development; Ewings sarcoma; FDA approved; Flow Cytometry; Gene Delivery; Gene Expression; Genomics; Glioma; Human; Immune; Immunocompetent; Immunologic Monitoring; Immunosuppression; Immunotherapy; Impairment; Inflammatory; Liposomes; Macrophage; Malignant Childhood Neoplasm; Measures; Mediating; Modeling; Monoclonal Antibody Therapy; Myeloid-derived suppressor cells; NK cell therapy; Natural Killer Cells; Neuroblastoma; Oncolytic; Oncolytic viruses; Phagocytes; Phagocytosis; Phenotype; Population; Relapse; Research Personnel; Resource Sharing; Rhabdomyosarcoma; Role; Shapes; Signal Transduction; Solid Neoplasm; Stress; Suppressor-Effector T-Lymphocytes; T cell clonality; T cell receptor repertoire sequencing; T-Lymphocyte; Testing; Treatment Efficacy; Tumor-associated macrophages; Tumor-infiltrating immune cells; Vaccines; Virotherapy; Virus; Virus Diseases; Work; Xenograft Model; Xenograft procedure; bisphosphonate; cancer cell; cancer immunotherapy; cancer regression; cancer therapy; cancer type; chemokine; chemotherapy; chimeric antigen receptor T cells; cytotoxic; exhaustion; gain of function; herpes virotherapy; innate immune mechanisms; loss of function; member; nanodiamond; neoplastic cell; neutrophil; new technology; osteosarcoma; pediatric patients; permissiveness; recruit; synergism; therapeutic target; time of flight mass spectrometry; trafficking; transcriptomics; tumor; tumor microenvironment; tumor xenograft; tumor-immune system interactions; vaccine strategy

Abstract - Project 4: Cure rates for pediatric patients with relapsed or metastatic solid tumors remain unacceptably low. Cancer immunotherapies hold great promise, but scores of disappointing studies highlight our relative ignorance in understanding the immunosuppressive microenvironment within solid tumors. Because of their central role in mediating immunosuppression, tumor associated macrophages (TAMs), typically “polarized” to a so-called M2- like immunosuppressive phenotype, and myeloid-derived suppressor cells (MDSC), are thought to be important therapeutic targets. We have found a clinically viable strategy that simultaneously reduces TAMs/MDSC (we dub “myelolytic”) and polarizes the microenvironment (via oncolytic virus infection), resulting in significant antitumor efficacy. We hypothesize that targeting TAM and MDSC by combining “myelolytic” therapies with pro- inflammatory therapies activates innate antitumor mechanisms that cause cancer regressions and reshapes the solid tumor microenvironment to be more permissive to cellular immunotherapies. In aim 1, we will determine the mechanism(s) by which combined myelolytic-virotherapy drives tumor regressions. We will use novel technologies such as fluorescent nanodiamonds to determine effects on innate immune cell phagocytosis of tumor cells. We will utilize the Genomics & Immune Monitoring Shared Resource Core B directed by Dr. Elaine Mardis to conduct flow cytometry with time-of-flight mass spectrometry and single cell transcriptomics to determine the effects on immune cell composition and polarization. We will also utilize gain- and loss-of-function approaches to determine if loss of MDSC are critical for enabling tumor regressions with myelolytic-virotherapy. We will also test combination therapies in xenograft and immunocompetent models of other cancer types to confirm its generalizability (osteosarcoma, Ewing sarcoma, rhabdomyosarcoma, neuroblastoma). In aim 2, we will determine the effects of myelolytic-virotherapy on T cell-mediated immunotherapies. We will examine the effect of myelolysis alone and combined with virotherapy on the efficacy of antitumor T cells in a T cell exhaustion setting and with CAR-T cells (with Project 1 Leader Dean Lee and co- investigator Ruoning Wang, PI-DDN U01 member). We will work with Core B to examine the effects on T cell clonality using TCR sequencing. In aim 3, we will determine whether combined myelolytic-virotherapy enhances the efficacy of NK-based cellular therapies. We will work with Project 1 Leader Dean Lee and Project 2 Leader Mitch Cairo to study the effects on adoptive NK and CAR-NK cell therapy. Overall, with this project we will further elucidate, test and develop strategies to modulate the tumor microenvironment to facilitate innate immune cells as cancer therapy. Our findings may be applicable across a broad panel of pediatric cancer types and thus fits well into the aims of the Pediatric Immunotherapy Discovery and Development Network.

摘要 - 项目 4： 复发或转移性实体瘤儿科患者的治愈率仍然低得令人无法接受。癌症 免疫疗法前景广阔，但大量令人失望的研究凸显了我们对免疫疗法的相对无知。 了解实体瘤内的免疫抑制微环境。由于他们在其中的核心作用 介导免疫抑制的肿瘤相关巨噬细胞 (TAM)，通常“极化”为所谓的 M2- 像免疫抑制表型和骨髓源性抑制细胞（MDSC）一样，被认为很重要 治疗目标。我们发现了一种临床上可行的策略，可以同时减少 TAM/MDSC（我们称之为 “溶髓”）并使微环境极化（通过溶瘤病毒感染），从而产生显着的抗肿瘤作用 功效。我们假设通过将“溶髓”疗法与促骨髓基质细胞疗法相结合来靶向 TAM 和 MDSC 炎症疗法激活先天抗肿瘤机制，导致癌症消退和 重塑实体瘤微环境，使其更适合细胞免疫疗法。瞄准目标 1，我们将确定联合溶髓病毒疗法驱动肿瘤的机制 回归。我们将使用荧光纳米金刚石等新技术来确定对先天的影响 免疫细胞对肿瘤细胞的吞噬作用。我们将利用基因组学和免疫监测共享资源 Core B 由 Elaine Mardis 博士指导，采用飞行时间质谱和单通道流式细胞术进行分析 细胞转录组学以确定对免疫细胞组成和极化的影响。我们还将利用 确定 MDSC 丧失是否对于实现肿瘤消退至关重要的功能获得和丧失方法 与溶髓病毒疗法。我们还将在异种移植和免疫活性模型中测试联合疗法 其他癌症类型以确认其普遍性（骨肉瘤、尤文肉瘤、横纹肌肉瘤、 成神经细胞瘤）。在目标 2 中，我们将确定溶髓病毒疗法对 T 细胞介导的作用 免疫疗法。我们将检查单独的骨髓溶解和联合病毒疗法对疗效的影响 在 T 细胞耗竭环境中和使用 CAR-T 细胞对抗肿瘤 T 细胞进行研究（与项目 1 负责人 Dean Lee 和合作伙伴一起） 研究员王若宁，PI-DDN U01 成员）。我们将与Core B合作检查对T细胞的影响 使用 TCR 测序进行克隆性分析。在目标 3 中，我们将确定是否联合溶髓病毒疗法 增强基于 NK 的细胞疗法的功效。我们将与项目 1 负责人 Dean Lee 和 项目 2 负责人 Mitch Cairo 负责研究过继性 NK 和 CAR-NK 细胞疗法的影响。总的来说，有了这个 该项目我们将进一步阐明、测试和开发调节肿瘤微环境的策略，以促进 先天免疫细胞作为癌症治疗。我们的研究结果可能适用于广泛的儿科癌症 类型，因此非常适合儿科免疫疗法发现和开发网络的目标。