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）是致命的小儿实体瘤。生存率<50％，而生存患者的生存率为 许多与治疗相关的毒性，强调对新型肿瘤靶向疗法的迫切需求。 NB患者的生存 通过添加抗GD2抗体Dinutuximab改进，增加了对其他的兴趣 免疫疗法，例如NB细胞疗法的适应性转移。迄今为止，细胞疗法集中于αβT 细胞和NK细胞，迄今为止，这些细胞对实体瘤均无成功。伽马三角洲（γδ）T细胞 是一种创新和优越的选择，因为它们独立于MHC，直接对肿瘤细胞的细胞毒性，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 αβT细胞的同质性。由于EX不足 体内扩展方法。我们已经开发了符合GMP符合GMP的无血清制造策略 将γδT细胞扩展到足够的人类使用水平。扩展的细胞高度表达CD16，直接 参与抗体定向细胞毒性（ADCC）。我们已经表明γδT细胞从NB扩展 患者有效地杀死了NB细胞系并增强了Dinutuximab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启用数据，以证明我们γδT细胞产物的有效性。 我们还产生了两个靶向GD2的嵌合抗原受体（CAR），以将γδT细胞定位到NB，并且 表明与非修饰细胞相比，CAR修饰的免疫能力细胞具有增强的细胞毒性。 我们还制定了与正常免疫细胞的化学疗法抗性的策略。这导致了我们 创新的“耐药免疫疗法”平台，通过该平台，可以共同管理化学保护的γδT细胞 通过化学疗法可显着提高抗NB效率。因此，我们的第二个目标是基因 使用一种新型的重组病毒载体的cDNA序列的γδT细胞来编码 甲基鸟氨酸甲基转移酶，MGMT，替莫唑胺（TMZ）抗性基因和我们的抗GD2型式甲烷。 将分析修饰的γδT细胞的各种遗传和功能特征以及抗肿瘤效力， 单独和使用TMZ，使用NB细胞系和患者在体外和体内衍生的异种移植物。目标 该提案的内容是1）开发可靠的临床前数据，以支持儿科使用自体的第一个 扩展的γδT细胞用于复发性NB，2）确定耐药性/GD2CARγδT细胞是否是优质 为未改性的γδT细胞提供信息，以告知未来的临床研究。这个免疫疗法平台在NB的成功将 为使用类似的策略提供基础来治疗其他癌症。