Simultaneous Targeting of Tumor and Stroma Cells to Enhance Solid Tumor CAR-T Cell Therapy

同时靶向肿瘤和基质细胞以增强实体瘤 CAR-T 细胞治疗

• 批准号: 10156815
• 负责人: Nicole J. Shirkey-Son
• 金额: $ 39.97万
• 依托单位: LUMINARY THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10156815
• 关键词: Address; Adoptive Transfer; Antigen Targeting; Antigens; Architecture; B-Cell Leukemia; Binding; Biological Assay; Biotechnology; CAR T cell therapy; CD19 gene; Carcinoma; Cell physiology; Cells; Childhood Acute Lymphocytic Leukemia; Coculture Techniques; DHFR gene; DNA Sequence; DNA Transposons; Desmoplastic; Development; Engineering; FDA approved; Fibroblasts; Flow Cytometry; Generations; Genetic; Harvest; Human; Immune; Immunotherapeutic agent; Immunotherapy; In Vitro; Infiltration; MS4A1 gene; Malignant Neoplasms; Malignant neoplasm of pancreas; Measures; Mediating; Methotrexate; Monitor; Mus; Pancreatic carcinoma; Patient-Focused Outcomes; Phase; Plague; Population; Process; Proteins; Publishing; Recurrence; Resistance; Resistance development; Small Business Innovation Research Grant; Solid Neoplasm; Specificity; System; T cell response; T cell therapy; T-Cell Activation; T-Cell Development; T-Lymphocyte; Testing; Therapeutic; Tissues; Tumor Burden; Tumor-infiltrating immune cells; United States National Institutes of Health; Weight; Work; Xenograft Model; Xenograft procedure; base; cancer biomarkers; cancer type; cell stroma; chimeric antigen receptor; chimeric antigen receptor T cells; clinically relevant; commercial application; cytotoxic; engineered T cells; exhaustion; experience; expression vector; fibroblast-activating factor; flexibility; improved; improved outcome; in vitro Assay; in vivo; ineffective therapies; leukemia/lymphoma; mesothelin; mouse model; mutant; neoplastic cell; new technology; non-viral gene delivery; novel; novel therapeutics; pancreatic cancer model; preclinical study; prevent; receptor expression; response; stem; success; targeted cancer therapy; therapy resistant; transgene expression; tumor; tumor growth; tumor microenvironment

Abstract The use of T cells engineered to express specific chimeric antigen receptors (CARs) to treat cancer has generated durable cures for many types of cancer and resulted in the first FDA approved CAR-T cell therapy to treat childhood acute lymphoblastic leukemia in 2017. Despite this success, CAR-T immunotherapies have been much less effective at targeting solid tumors. Part of this limited success stems from the solid tumor microenvironment, which forms a physical barrier to immune cell infiltration and produces soluble factors that downregulate T cell activity and accelerate T cell exhaustion. While immunotherapies targeting solid tumors are initially effective, the tumor microenvironment’s inhibition of T cells prevents these treatments from producing durable responses. In this application, we propose a novel CAR-T cell therapy aimed to improve outcomes for patients with advanced stage pancreatic cancer by overcoming the deficiencies that plague current CAR-T cell therapies. To this end, we will engineer T cells to express multiple CARs, enabling these cells to target tumor cells and cells in the immune-suppressive tumor microenvironment. Specifically, we will leverage the non-viral, Tc Buster DNA transposon system to insert a large multicistronic genetic construct containing multiple CARs and a selection marker into T cells. Using this platform, we will generate T cells with CARs targeting mesothelin (MSLN), a protein expressed by 80-85% of pancreatic cancer tumors, and fibroblast activation protein (FAP), a marker of cancer associated fibroblasts in the tumor microenvironment. We will then select a pure population of T cells expressing MSLN- and FAP-CARs and determine the activity and specificity of these cells in vitro. We expect that engineered T cells will generate a specific and robust response, eliciting cytotoxic functions only against cells expressing their target antigen. We will then determine the efficacy of engineered T cells in vivo using a xenograft mouse model to generate MSLN and FAP positive pancreatic carcinomas followed by adoptive transfer of T cells. We expect immunotherapeutic delivery of bispecific T cells expressing FAP-CARs and MSLN- CARs will elicit a robust and long-lasting T cell response against MSLN+/FAP+ solid tumors resulting in tumor shrinkage and increased survival. Furthermore, we expect the development of a flexible, efficient, and reliable process to generate bispecific T cells with a single, non-viral gene delivery approach will facilitate the emergence of novel therapies to overcome many issues facing engineered T cell therapy today, including antigen escape and target specificity.

抽象的 使用经过改造可表达特定嵌合抗原受体 (CAR) 的 T 细胞来治疗癌症 为许多类型的癌症提供了持久的治疗方法，并导致第一个 FDA 批准的 CAR-T 细胞疗法 2017 年治疗儿童急性淋巴细胞白血病。尽管取得了这一成功，但 CAR-T 免疫疗法 这种有限成功的部分原因在于实体瘤。 微环境，形成免疫细胞渗透的物理屏障，并产生可溶性因子 下调 T 细胞活性并加速 T 细胞耗竭，而针对实体瘤的免疫疗法则相反。 最初有效，肿瘤微环境对 T 细胞的抑制阻止了这些治疗产生 在此应用中，我们提出了一种新型 CAR-T 细胞疗法，旨在改善治疗结果。 通过克服困扰当前 CAR-T 细胞的缺陷，治疗晚期胰腺癌患者 为此，我们将改造 T 细胞来表达多种 CAR，使这些细胞能够靶向肿瘤。 具体来说，我们将利用非病毒、 Tc Buster DNA 转座子系统可插入包含多个 CAR 的大型多顺反子遗传构建体 使用这个平台，我们将生成带有靶向间皮素的 CAR 的 T 细胞。 (MSLN)，一种由 80-85% 的胰腺癌肿瘤表达的蛋白质，以及成纤维细胞激活蛋白 (FAP)，一种 然后我们将选择肿瘤微环境中癌症相关成纤维细胞的纯群体。 T 细胞表达 MSLN-和 FAP-CAR，并在体外确定这些细胞的活性和特异性。 预计工程化 T 细胞将产生特异性且强大的反应，仅引发细胞毒性功能 然后，我们将确定工程化 T 细胞在体内的功效。 使用异种移植小鼠模型产生 MSLN 和 FAP 阳性胰腺癌，然后过继 我们期望表达 FAP-CAR 和 MSLN- 的双特异性 T 细胞进行免疫治疗递送。 CAR 将引发针对 MSLN+/FAP+ 实体瘤的强大而持久的 T 细胞反应，从而导致肿瘤 此外，我们期望开发一种灵活、高效、可靠的解决方案。 使用单一非病毒基因传递方法生成双特异性 T 细胞的过程将促进出现 克服当今工程 T 细胞疗法面临的许多问题的新疗法，包括抗原逃逸 和目标特异性。