Optimization of TIL Cell Manufacturing for Cancer Treatment

用于癌症治疗的 TIL 细胞制造优化

• 批准号: 10696746
• 负责人: Soon Seog Jeong
• 金额: $ 40万
• 依托单位: HUMAN CELL CO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696746
• 关键词: Adoptive Cell Transfers; Allogenic; Antibodies; Antigen Targeting; Antigens; Autologous; Autologous Tumor-Infiltrating Lymphocyte; Back; Biopsy; CD3 Antigens; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Center; Cell Line; Cells; Clinical; Clone Cells; Data; Diameter; Effectiveness; Endotoxins; Enrollment; Escherichia coli; Frequencies; Generations; Guidelines; Harvest; Human; IL17 gene; Immune checkpoint inhibitor; Immunotherapy; In Vitro; Individual; Infiltration; Interleukin-2; Letters; Link; Lymphocyte; Lymphocyte Activation; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Memory; Metastatic Melanoma; Monoclonal Antibodies; Mus; Operative Surgical Procedures; Pancreas; Pathway interactions; Patients; Phase I Clinical Trials; Phenotype; Population; Portugal; Privatization; Process; Production; Protocols documentation; Recombinants; Recurrence; Regulatory T-Lymphocyte; Relapse; Reporting; Resectable; Resected; Site; Solid Neoplasm; Source; Specimen; Sterility; T cell response; T memory cell; T-Lymphocyte; T-cell receptor repertoire; Therapeutic; Time; Tissues; Transfection; Tumor Antigens; Tumor Immunity; Tumor Promotion; Tumor Tissue; Tumor-Infiltrating Lymphocytes; Work; anti-CTLA4; anti-PD-1; anti-PD-L1; antigen-specific T cells; cancer therapy; cancer type; clinically significant; cytokine; effector T cell; efficacy validation; exhaust; experience; falls; fitness; improved; in vivo; innovation; lymphocyte product; manufacture; manufacturing process; neoantigens; objective response rate; pancreatic cancer patients; pancreatic neoplasm; protein purification; response; scale up; stem; stem cells; success; treatment optimization; tumor; tumor progression

ABSTRACT Adoptive cell therapy (ACT) using autologous tumor-infiltrating lymphocytes (TIL) has demonstrated tremendous potential for treatment of advanced solid tumors. Objective response rates ranging from 34 to 72% have been reported in patients with metastatic melanoma, with durable, complete tumor regression observed in up to 20% of treated patients. However, the current process for isolating, identifying, and expanding therapeutic TIL cells was established more than 30 years ago. TILs are stimulated with a murine anti-CD3 monoclonal antibody (OKT-3), high concentration of recombinant IL-2 produced from E. coli, and irradiated allogeneic or autologous feeder cells. Shortcomings of this process include the need for a surgically resectable tumor as a source of TIL cells, inability to grow TILs for a significant portion of patients, significant presence of regulatory T cells, and long production time. Moreover, TILs are predominantly differentiated into “old” effector T cells in vitro with a terminal phenotype, thereby reducing their long-term survival and antitumor effectiveness in vivo. Younger phenotype T cells, including stem cell memory and central memory T cells, provide superior persistence and antitumor immunity compared with effector memory T cells and effector T cells. This is consistent with recent clinical findings by Dr. Rosenberg and his group that the response of TILs against human cancer is primarily mediated by neoantigen-specific and stem-like CD8+ T cells (CD39-CD69-).Moreover, there is a high unmet need for rapidly progressing cancer types where the window of treatment is limited and where the time for TIL These shortcomings can be surmounted by improving the antibodies and cytokines used ex vivo and optimizing the combination and manufacturing process to robustly and rapidly produce TIL cells, thus, enabling TIL treatment for a broad spectrum of solid tumor patients with higher response rate and curative potential. production becomes of paramount importance. We have been using stably transfected HEK293 cells to produce proprietary antibodies and cytokines. These ancillary materials are critical in TIL manufacturing but are not intended to be part of the final cell product. O ur innovative products for TIL cell production as ex vivo therapeutics have demonstrated striking advantages over current commercial products for TIL production by improving the culture success rate, absolute expansion number, fitness and, critically, shortening the duration of TIL manufacturing while minimizing regulatory T cells. W e have formed a strategic partnership to rigorously evaluate the products and optimize the manufacturing process of TILs. Currently, TILs have been successfully cultured from small tissues of 60 pancreatic and 20 non- pancreatic tumors and scaled up using the Cocoon® Platform (Lonza) for pancreatic tumors, the most difficult TILs to grow so far. Critically, the TILs manufactured in clinical scale has a high frequency of CD8+CD39- CD69- T cells and the reactivities of TILs against neo-antigens were robustly detected by IFN release. The data obtained thus far show a focused, yet diverse TCR repertoire. In addition to a more general TCR analysis in TIL, we were able to link individual TCR clonotypes to individual private target antigens and to trace these back to the TIL product and to the corresponding harvested tumor tissue, respectively. Specific Aim. To determine whether anti-CD137HC and/or IL-12HC enriches antigen-specific T cells and anti- TGFHC, anti-IL-6HC and/or anti-IL-23HC blunts Th17 differentiation and IL-17 release; select top Expi293 cell clones and complete pilot scale production of proprietary antibodies and cytokines critical for TIL manufacturing. The strategic collaborator will validate the efficacy and consistency of our products and pursue regulatory clearance for clinical manufacturing of TILs from pancreatic patients. Importantly, a Phase 1 clinical trial for metastatic or recurrent pancreatic cancer patients has been planned, the first patient is anticipated to be enrolled as soon as 2023.

抽象的 使用自体肿瘤浸润淋巴细胞（TIL）的过继细胞疗法（ACT）已证明 治疗晚期实体瘤的巨大潜力。客观反应率范围为 34% 至 72% 据报道，在转移性黑色素瘤患者中观察到肿瘤持久、完全消退 高达 20% 的接受治疗的患者。然而，目前分离、鉴定和扩大治疗的过程 TIL细胞的建立已有30多年的历史。 TILs 用鼠抗 CD3 单克隆抗体刺激 抗体 (OKT-3)、大肠杆菌产生的高浓度重组 IL-2 以及经过辐照的同种异体或 自体饲养细胞。该过程的缺点包括需要通过手术切除肿瘤作为 TIL 细胞来源、大部分患者无法培养 TIL、显着存在调节性 T 细胞，生产时间长。此外，TIL 在体外主要分化为“旧”效应 T 细胞 具有终末表型，从而降低它们的长期存活和体内抗肿瘤效果。雅戈尔 表型 T 细胞，包括干细胞记忆和中央记忆 T 细胞，提供卓越的持久性和 与效应记忆 T 细胞和效应 T 细胞相比，抗肿瘤免疫。这与近期一致 Rosenberg 博士及其团队的临床发现表明，TIL 对人类癌症的反应主要是 由新抗原特异性和干细胞样 CD8+ T 细胞 (CD39-CD69-) 介导。此外，还有很高的未满足 需要快速进展的癌症类型，其中治疗窗口有限且 TIL 时间有限 这些缺点可以通过改进 体外使用抗体和细胞因子，并优化组合和制造工艺，以稳健地 并快速产生 TIL 细胞，从而使 TIL 治疗能够广泛用于患有以下疾病的实体瘤患者： 更高的缓解率和治疗潜力。 生产变得至关重要。 我们一直在使用稳定转染的 HEK293 细胞来生产专有抗体和细胞因子。这些 辅助材料在 TIL 制造中至关重要，但并不打算成为最终电池产品的一部分。 氧 你的 用于 TIL 细胞生产的创新产品作为离体疗法已表现出显着的优势 目前用于TIL生产的商业产品通过提高培养成功率，绝对扩张 数量、适应性，最重要的是，缩短 TIL 制造的持续时间，同时最大限度地减少调节性 T 细胞。 瓦 我们建立了战略合作伙伴关系，严格评估产品并优化制造 TIL 的过程。现在， TIL 已成功从 60 个胰腺小组织和 20 个非胰腺小组织中培养出来。 胰腺肿瘤和 使用 Cocoon® 平台 (Lonza) 扩大治疗胰腺肿瘤的规模，这是最困难的 TIL 到目前为止仍将增长。至关重要的是，临床规模生产的 TIL 具有高频率的 CD8+CD39- CD69-T 细胞和 TIL 对抗新抗原的反应性可通过 IFNγ 释放得到有力检测。数据 迄今为止获得的结果显示了一个集中但多样化的 TCR 库。除了 TIL 中更一般的 TCR 分析之外， 我们能够将个体 TCR 克隆型与个体私有靶抗原联系起来，并将其追溯到 TIL产物和相应的收获的肿瘤组织，分别。 具体目标。确定抗 CD137HC 和/或 IL-12HC 是否富集抗原特异性 T 细胞和抗 TGFβHC、抗 IL-6HC 和/或抗 IL-23HC 减弱 Th17 分化和 IL-17 释放；选择顶部 Expi293 单元格 克隆并完成对 TIL 制造至关重要的专有抗体和细胞因子的中试规模生产。 战略合作者将验证我们产品的功效和一致性，并寻求监管 获得胰腺患者 TIL 临床生产的许可。重要的是，一期临床试验 转移性或复发性胰腺癌患者已计划入组，预计将入组第一名患者 最快到 2023 年。