Exploiting chemoimmunotherapy strategies with genetically-engineered gd T cells

利用基因工程 gd T 细胞进行化学免疫治疗策略

• 批准号: 9585676
• 负责人: Kelly C Goldsmith
• 金额: $ 20.06万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9585676
• 关键词: Adoptive Transfer; Animals; Antibodies; Antibody Therapy; Antigens; Autologous; Autologous Stem Cell Transplantation; Biological Response Modifier Therapy; Biomedical Engineering; Cattle; Cell Death; Cell Line; Cell Therapy; Cell-Mediated Cytolysis; Cells; Cellular immunotherapy; Characteristics; Child; Childhood Extracranial Solid Tumor; Childhood Solid Neoplasm; Clinical; Clinical Research; Complementary DNA; Cyclic GMP; Data; Drug resistance; Effectiveness; Engineering; Evaluation; FCGR3B gene; FDA approved; Fc Receptor; Foundations; Future; Gene-Modified; Genetic; Genetic Engineering; Glioblastoma; Goals; Human; Immune; Immune Targeting; Immuno-Chemotherapy; Immunocompetent; Immunotherapy; In Vitro; Individual; Inflammatory; MGMT gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Methods; Methyltransferase; Modeling; Monoclonal Antibodies; Mus; Natural Killer Cells; Neoplasm Metastasis; Neuroblastoma; Newly Diagnosed; Patients; Pediatrics; Pharmacology and Toxicology; Property; Reagent; Recombinant Proteins; Recurrence; Regimen; Relapse; Resistance; Risk; Serum; Serum-Free Culture Media; Solid Neoplasm; Stress; Surface; T cell therapy; T-Lymphocyte; Testing; Toxic effect; Treatment-related toxicity; Tretinoin; Up-Regulation; Xenograft procedure; anti-cancer; antibody-dependent cell cytotoxicity; base; bisphosphonate; cDNA Expression; chemotherapy; chimeric antigen receptor; clinical application; clinically translatable; cytokine; cytotoxic; cytotoxicity; disorder risk; high risk; human model; improved; in vivo; in vivo evaluation; innovation; interest; neoplastic cell; neuroblastoma cell; novel; novel therapeutics; objective response rate; pre-clinical; recombinant viral vector; research clinical testing; resistance gene; standard of care; success; targeted treatment; temozolomide; treatment strategy; tumor; tumor microenvironment; γδ T cells

Project Summary High-risk neuroblastoma (HR NB) is a lethal pediatric solid tumor. Survival is < 50% and those that survive suffer many treatment related toxicities, stressing a critical need for novel tumor- targeted therapies. NB patient survival improved with the addition of the anti-GD2 antibody, dinutuximab, increasing the interest in other immunotherapies like adoptive transfer of cellular therapies for NB. Cellular therapy has so far focused on αβ T cells and NK cells, which to date have been uniformly unsuccessful for solid tumors. Gamma delta (γδ) T cells are an innovative and superior choice as they are MHC independent, directly cytotoxic to tumor cells, including NB, can recognize and target immune-suppressing cells in the tumor microenvironment, and lack the alloreactivity of αβ T cells. Adoptive γδ T cells have not been widely used clinically to date due to inadequate ex vivo expansion methods. We have developed a GMP-compliant serum free manufacturing strategy to expand γδ T cells to sufficient levels for human use. Expanded cells highly express CD16, which is directly involved in antibody directed cellular cytotoxicity (ADCC). We have shown that γδ T cells expanded from NB patients effectively kill NB cell lines and enhance dinutuximab-induced NB cell death. Therefore, our primary objective is to generate preclinical and IND enabling data demonstrating the effectiveness of our γδ T cell product. We have also generated two chimeric antigen receptors (CARs) targeting GD2 to localize γδ T cells to NB, and show that CAR-modified immunocompetent cells have enhanced cytotoxicity compared to non-modified cells. We have also developed strategies to confer chemotherapy resistance to normal immune cells. This led to our innovative “drug resistant immunotherapy” platform whereby chemo-protected γδ T cells can be co-administered with chemotherapy to significantly augment anti-NB efficacy. Our second objective is therefore to genetically engineer γδ T cells using a novel recombinant viral vector-based delivery of cDNA sequences that encode for methylguanine methyltransferase, MGMT, a temozolomide (TMZ) resistance gene and our anti-GD2-CAR. Modified γδ T cells will be analyzed for various genetic and functional characteristics and anti-tumor potency, both alone and with TMZ, using NB cell lines and patient derived xenografts both in vitro and in vivo. The goals of this proposal are to 1) develop robust preclinical data to support the first in pediatrics use of autologous expanded γδ T cells for recurrent NB, and 2) determine whether drug resistant/GD2CAR γδ T cells are superior to unmodified γδ T cells to inform future clinical studies. Success of this immunotherapy platform in NB will provide the foundation to treat other cancers using a similar strategy.

项目摘要 高危神经母细胞瘤（HR NB）是一种致命的儿科实体瘤。存活率< 50%，存活者遭受痛苦 许多治疗相关的毒性，强调了对新的肿瘤靶向治疗的迫切需要。NB患者生存期 随着抗GD 2抗体dinutuximab的加入， 免疫疗法，如用于NB的细胞疗法的过继转移。迄今为止，细胞疗法主要集中在αβ T 细胞和NK细胞，迄今为止，它们对于实体瘤都是不成功的。γ δ T细胞 是一种创新的上级选择，因为它们不依赖于MHC，对肿瘤细胞具有直接细胞毒性，包括 NB可以识别和靶向肿瘤微环境中的免疫抑制细胞，并且缺乏免疫抑制细胞。 αβ T细胞的同种异体反应性。由于缺乏足够的表达，过继性γδ T细胞迄今为止尚未在临床上广泛使用 体内扩增方法。我们已经开发了符合GMP的无血清生产策略， 将γδ T细胞扩增至足以供人类使用的水平。扩增的细胞高度表达CD 16，其直接表达CD 16。 抗体介导的细胞毒性（ADCC）。我们已经表明，γδ T细胞从NB扩增， 患者有效地杀死NB细胞系并增强地奴昔单抗诱导的NB细胞死亡。因此，我们的主要 目的是生成临床前和IND支持数据，证明我们的γδ T细胞产品的有效性。 我们还产生了两种靶向GD 2的嵌合抗原受体（汽车），以将γδ T细胞定位于NB， 显示CAR修饰的免疫活性细胞与未修饰的细胞相比具有增强的细胞毒性。 我们还开发了赋予正常免疫细胞化疗耐药性的策略。这导致了我们的 创新的“耐药免疫疗法”平台，其中可以共同施用化学保护的γδ T细胞 化疗以显著增强抗NB功效。因此，我们的第二个目标是从基因上 使用新型的基于重组病毒载体的cDNA序列递送工程化γδ T细胞，所述cDNA序列编码 甲基鸟嘌呤甲基转移酶、MGMT、替莫唑胺（TMZ）抗性基因和我们的抗GD 2-CAR。 将分析修饰的γδ T细胞的各种遗传和功能特征以及抗肿瘤效力， 单独和与TMZ一起，使用NB细胞系和患者来源的异种移植物在体外和体内进行。的目标 该提案的目的是：1）开发可靠的临床前数据，以支持首次在儿科使用自体 用于复发性NB的扩增的γδ T细胞，和2）确定药物抗性/GD 2 CAR γδ T细胞是否是上级的 未修饰的γδ T细胞，为未来的临床研究提供信息。这种免疫治疗平台在NB中的成功将 为使用类似策略治疗其他癌症提供了基础。