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.

摘要 使用自体肿瘤浸润性淋巴细胞（TIL）的免疫细胞疗法（ACT）已经证明， 治疗晚期实体瘤的巨大潜力。客观缓解率范围为34%至72% 已在转移性黑色素瘤患者中报告， 高达20%的患者接受治疗。然而，目前用于分离、鉴定和扩大治疗的方法， TIL细胞是30多年前建立的。用鼠抗CD 3单克隆抗体刺激TIL， 抗体（OKT-3）、高浓度重组IL-2。大肠杆菌，和辐射同种异体或 自体饲养细胞这一过程的缺点包括需要通过手术切除的肿瘤作为治疗方法。 TIL细胞来源，大部分患者不能生长TIL，大量存在调节性T细胞， 细胞，生产时间长。此外，TIL在体外主要分化为“老的”效应T细胞， 具有终末表型，从而降低它们的长期存活率和体内抗肿瘤效力。年轻 表型T细胞，包括干细胞记忆和中枢记忆T细胞，提供上级的持久性和 与效应记忆T细胞和效应T细胞相比的抗肿瘤免疫。这与最近 Rosenberg博士和他的小组的临床研究发现，TIL对人类癌症的反应主要是 由新抗原特异性和干细胞样CD 8 + T细胞（CD 39-CD 69-）介导。 需要快速进展的癌症类型，其中治疗窗口有限，并且其中TIL的时间 这些缺点可以通过改进 体外使用的抗体和细胞因子，并优化组合和制造过程， 并快速产生TIL细胞，从而使TIL治疗能够用于具有以下特征的广谱实体瘤患者： 更高的反应率和治愈潜力。 生产变得至关重要。 我们一直在使用稳定转染的HEK 293细胞来生产专有抗体和细胞因子。这些 辅助材料在TIL制造中是关键的，但不打算成为最终细胞产品的一部分。 O ur 作为离体治疗剂的用于TIL细胞生产的创新产品已经显示出显著的优势， 目前商品化的TIL产品生产通过提高培养成功率，绝对扩产 数量、适应性，并且关键地，缩短TIL制造的持续时间，同时最小化调节性T细胞。 W 我们已经建立了战略合作伙伴关系，严格评估产品并优化制造 TILs的过程。目前， TILs已经成功地从60个胰腺和20个非胰腺的小组织中培养出来。 胰腺肿瘤， 使用Cocoon®平台（Lonza）扩大规模用于胰腺肿瘤，这是最困难的 才能长到现在。重要的是，以临床规模生产的TIL具有高频率的CD 8 + CD 39-T细胞亚群。 CD 69- T细胞和TIL对新抗原的反应性通过IFN γ释放被稳健地检测。数据 迄今为止获得的TCR库显示出集中的但多样的TCR库。除了TIL中更一般的TCR分析之外， 我们能够将单个TCR克隆型与单个私人靶抗原联系起来，并将其追溯到 TIL产物和相应的收获的肿瘤组织。 具体目标。为了确定抗-CD 137 HC和/或IL-12 HC是否富集抗原特异性T细胞和抗-CD 137 HC和/或IL-12 HC是否富集抗原特异性T细胞， TGF β HC、抗IL-6 HC和/或抗IL-23 HC减弱Th 17分化和IL-17释放;选择最佳Expi 293细胞 克隆和完整的中试规模生产专有的抗体和细胞因子的关键TIL制造。 战略合作者将验证我们产品的有效性和一致性，并寻求监管 用于从胰腺患者临床生产TIL的许可。重要的是，一项1期临床试验， 已计划入组转移性或复发性胰腺癌患者，预计将入组首例患者 到2023年。