Genome Engineered Natural Killer Cell Immunotherapy against Human Osteosarcoma

针对人类骨肉瘤的基因组工程自然杀伤细胞免疫疗法

基本信息

批准号：
10670762
负责人：
Gabrielle Matilde Robbins
金额：
$ 5.13万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-16 至 2025-08-15
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10670762
关键词：
Activated Natural Killer Cell Allogenic Antibodies Antigen Targeting Antigens Autoimmunity Binding Biological Assay Bioluminescence Body Weight decreased CAR T cell therapy CBL gene CRISPR/Cas technology Cell Line Cell physiology Cell-Mediated Cytolysis Cells Clinical Clinical Trials Coculture Techniques Data Disease Dose ERBB2 gene Engineering Engraftment FCGR3A gene Flow Cytometry Generations Genes Genetic Code Genetic Engineering Genome engineering Genomics Goals Health Hematologic Neoplasms Human Immune Immune system Immunotherapy In Vitro Ink Interleukin-15 Interphase Cell Knock-out Label Laboratories Longevity Luciferases MADH3 gene Malignant Neoplasms Mediating Membrane Modality Modeling Monitor Mus Myeloproliferative disease NK Cell Activation Natural Killer Cell Immunotherapy Natural Killer Cells Patient-Focused Outcomes Patients Proteins Publications Reagent Recombinant adeno-associated virus (rAAV)Rest Safety Solid Neoplasm System T cell therapy T-Lymphocyte Testing Translating Treatment Efficacy Tumor Escape Tumor Suppression Western Blotting Work Xenograft Model antibody-dependent cell cytotoxicity bioluminescence imaging cancer cell cancer immunotherapy cell killing cell transformation chimeric antigen receptor chimeric antigen receptor T cells comparison control cytokine release syndrome cytotoxic cytotoxicity design engineered NK cell fighting graft vs host disease high risk improved in vivo in vivo evaluation induced pluripotent stem cell interest interleukin-21 knockout gene neoplastic cell neurotoxicity next generation sequencing novel osteosarcoma peripheral blood pre-clinical public health relevance receptor response site-specific integration success treatment group tumor tumor microenvironment

项目摘要

Abstract Over the last decade Chimeric Antigen Receptor based T cell therapy (CAR-T) has developed into an effective immunotherapy for some cancers. However, CAR-T cell therapies have several shortcomings and clinical success has primarily been limited to hematological cancers. Challenges of CAR-T cell therapy include tumor immune evasion through loss of target antigen expression by tumor cells and inhibition of CAR-T cell function by tumor expressed inhibitory molecules. Natural killer (NK) cells present an alternative to T cells that could be more effective due to their ability to perform both antigen dependent and independent killing. NK cells have demonstrated antigen specific killing when engineered to express T cell CARs and NK cells also mediate the direct killing of transformed cells with reduced or absent MHC expression. In fact, NK cells carry out antibody dependent cell mediated cytotoxicity (ACDD) of cells that bind antibodies via the NK cell CD16A receptor. Due to the multiple modalities for cancer cell killing, there is an increased interest in NK cells for cancer immunotherapy. As NK cells are not associated with graft versus host disease, neurotoxicity, long-term autoimmunity, nor cytokine release syndrome, they are more suited for use in allogeneic settings than T cells and have significant clinical potential for use as off-the-shelf products. However, previous publications and clinical trials have demonstrated that the use of unmanipulated NK cells to treat cancer is minimally effective, likely due to limited engraftment, little in vivo expansion, and suppression by the tumor microenvironment. NK cells activated and expanded with feeder cells expressing membrane bound interleukin-21 (mbIL-21) have shown promising results clinically with high-risk myeloid malignancies and preclinically in several solid tumor models. Therefore, we hypothesize that activated/expanded NK cells that have be genetically edited can be used to successfully treat osteosarcoma, a disease for which patient outcome has not improved in over thirty years. Our proposed objectives are to evaluate the baseline response of rested- and activated-NK cells against various osteosarcoma cell lines, knockout negative regulators of NK cell function (specifically, c-CBL, IL-1R8, and SMAD3), and implement a specific CAR that optimally activates NK cell antigen-specific killing. Genetically engineered NK cells will be evaluated for enhanced therapeutic efficacy and safety in osteosarcoma models. Our preliminary data strongly supports the hypothesis that NK cell-based cancer immunotherapy can be fully realized using activated, genome engineered NK cells.

抽象的在过去的十年中，基于嵌合抗原受体的 T 细胞疗法 (CAR-T) 已发展成为一种有效的治疗方法。针对某些癌症的免疫疗法。然而，CAR-T细胞疗法存在一些缺点，临床效果不佳。成功主要限于血液癌症。 CAR-T细胞疗法的挑战包括肿瘤通过肿瘤细胞丧失靶抗原表达和抑制 CAR-T 细胞功能来逃避免疫通过肿瘤表达的抑制分子。自然杀伤 (NK) 细胞是 T 细胞的替代品由于它们能够执行抗原依赖性和独立杀伤，因此更加有效。 NK 细胞有当被设计来表达 T 细胞 CAR 时，证明了抗原特异性杀伤，并且 NK 细胞也介导直接杀死 MHC 表达减少或缺失的转化细胞。事实上，NK细胞携带抗体通过 NK 细胞 CD16A 受体结合抗体的细胞的依赖性细胞介导的细胞毒性 (ACDD)。到期的由于多种杀死癌细胞的方式，人们对 NK 细胞治疗癌症的兴趣日益浓厚免疫疗法。由于 NK 细胞与移植物抗宿主病、神经毒性、长期自身免疫，也不是细胞因子释放综合征，它们比 T 细胞更适合在同种异体环境中使用并具有作为现成产品使用的巨大临床潜力。然而，之前的出版物和临床试验表明，使用未经操作的 NK 细胞治疗癌症效果甚微，可能是由于植入有限、体内扩张很少以及肿瘤微环境的抑制。 NK细胞用表达膜结合白细胞介素 21 (mbIL-21) 的饲养细胞激活和扩增的细胞具有在高风险骨髓恶性肿瘤的临床和几种实体瘤的临床前研究中显示出有希望的结果模型。因此，我们假设经过基因编辑的激活/扩增的 NK 细胞可以用于成功治疗骨肉瘤，这种疾病的患者治疗效果在三十多年来一直没有改善年。我们提出的目标是评估静息和激活 NK 细胞的基线反应各种骨肉瘤细胞系，敲除 NK 细胞功能的负调节因子（特别是 c-CBL、IL-1R8、和 SMAD3），并实现最佳激活 NK 细胞抗原特异性杀伤的特定 CAR。遗传上将评估工程化 NK 细胞在骨肉瘤模型中增强的治疗功效和安全性。我们的初步数据有力地支持了这样的假设：基于 NK 细胞的癌症免疫疗法可以完全使用激活的基因组工程 NK 细胞实现。