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一些癌症的免疫疗法。但是，CAR-T细胞疗法有几个缺点和临床成功主要仅限于血液学癌症。 CAR-T细胞疗法的挑战包括肿瘤通过肿瘤细胞失去靶抗原表达和抑制CAR-T细胞功能的免疫逃避肿瘤表达抑制分子。天然杀手（NK）细胞提出了T细胞的替代方法由于它们具有执行抗原依赖性和独立杀害的能力，因此更有效。 NK细胞具有在设计以表达T细胞汽车和NK细胞的工程时，证明了抗原特异性杀戮也可以介导直接杀死转化的细胞，并降低或不存在MHC表达。实际上，NK细胞进行抗体通过NK细胞CD16A受体结合抗体的细胞的依赖细胞介导的细胞毒性（ACDD）。到期的对于癌细胞杀死的多种方式，对NK细胞对癌症的兴趣增加了免疫疗法。由于NK细胞与移植物与宿主疾病无关，因此神经毒性，长期自身免疫性和细胞因子释放综合征，它们比T细胞更适合于同种异体设置并具有用作现成产品的巨大临床潜力。但是，以前的出版物和临床试验表明，使用无操纵的NK细胞治疗癌症是最小有效的，可能是由于植入有限，体内膨胀很少以及肿瘤微环境抑制。 NK 用表达膜结合白细胞介素21（MBIL-21）激活和扩展的细胞被激活和扩展通过高风险的髓样恶性肿瘤在临床上显示出令人鼓舞的结果型号。因此，我们假设已遗传编辑的活化/扩展的NK细胞可以是用于成功治疗骨肉瘤年。我们提出的目标是评估静止的NK和激活的NK细胞的基线响应各种骨肉瘤细胞系，NK细胞功能的基因敲除负调节剂（特别是C-CBL，IL-1R8，和SMAD3），并实施特定的汽车，该汽车可最佳地激活NK细胞抗原特异性杀戮。遗传将评估工程的NK细胞在骨肉瘤模型中的治疗功效和安全性增强。我们的初步数据强烈支持以下假设：基于NK细胞的癌症免疫疗法可以完全使用活化的基因组工程的NK细胞实现。