Cytokine Delivery By Tumor-Seeking Lymphocytes

肿瘤寻找淋巴细胞的细胞因子传递

• 批准号: 6865438
• 负责人: Per H. Basse
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-02 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6865438
• 关键词: Adenoviridae; Retroviridae; biotechnology; chemokine; cytokine; cytotoxic T lymphocyte; gene delivery system; gene expression; gene therapy; genetic transduction; genetically modified animals; interleukin 2; laboratory mouse; lymphocyte; natural killer cells; neoplasm /cancer genetics; neoplasm /cancer immunology; neoplasm /cancer immunotherapy; neoplasm /cancer therapy; passive immunization; therapy design /development; transfection /expression vector

DESCRIPTION (provided by applicant): Targeted delivery to tumors of cytokine genes capable of initiating anti-tumor immune responses is a promising approach for cancer therapy. However, success has been limited due to the lack of efficient methods to ensure gene-expression selectively in established tumors. We have recently shown that adoptively transferred, IL2-activated natural killer (A-NK) cells and PHA- and IL-2-stimulated CD8+ T lymphocytes (called T-LAK cells) localize into tumor with impressive selectively. Our goal is to take advantage of this skill, and demonstrate that cytokine gene-transduced, tumor-seeking A-NK and/or T-LAK cells can be used as vehicles for the delivery of anti-cancer cytokines selectively to tumors and to show that this forced cytokine expression in the tumors will lead to strong anti-tumor immune responses. The SPECIFIC AIMS of this 3-year project are to: 1) Demonstrate that A-N K cells can be transduced with adenoviral vectors to produce significant amounts of cytokines; 2) Analyze the ability of cytokine gene-transduced T-LAK and A-NK cells to localize into tumors and to produce cytokines at the tumor site, and; 3) Demonstrate that adoptive transfer of cytokine gene-transduced A-NK and T-LAK cells leads to elimination of tumors without induction of systemic toxicity. To accomplish this, T-LAK and A-NK cells will be adeno- or retro-virally transduced with one or several genes encoding for cytokines/chemokines known to have anti tumor effect via stimulation of either NK cells, DCs, and T helper cells and CTLs, namely IL-2, IL-4, IL12, IL-15, IL-18, IFNg, GM-CSF, RANTES and lymphotactin. The ability of successfully transduced T-LAK and A-NK cells to localize into tumors and to produce cytokine at the tumor site will be tested in animal tumor models. The host response, such as increased accumulation of effector cells into the tumor tissue, induced by the intratumorally produced cytokines, will be measured. The therapeutic potential of this approach will be evaluated by analyzing tumor-reduction resulting from injection of batches of A-NK and/or T-LAK cells, each capable of producing one or several anti-cancer cytokines. The mechanism behind any anti-tumor effect observed, will be elucidated. Depending on the cytokines produced by the tumor-seeking A-NK and T-LAK cells, we anticipate that we can manipulate the host immune system to elicit strong innate and adaptive anti-tumor responses without induction of toxic side effects. On this background, it should be possible to substantially improve the efficacy of tumor-targeted cytokine gene-therapy of cancer.

描述（由申请人提供）：靶向递送能够引发抗肿瘤免疫应答的细胞因子基因至肿瘤是一种有前景的癌症治疗方法。然而，由于缺乏有效的方法来确保基因在已建立的肿瘤中选择性表达，成功受到限制。我们最近发现过继转移的IL-2激活的自然杀伤（A-NK）细胞和PHA和IL-2刺激的CD 8 + T淋巴细胞（称为T-LAK细胞）具有显著的选择性定位于肿瘤。我们的目标是利用这种技术，并证明细胞因子基因转导的肿瘤寻求性A-NK和/或T-LAK细胞可用作将抗癌细胞因子选择性递送至肿瘤的载体，并显示这种强制的细胞因子在肿瘤中的表达将导致强烈的抗肿瘤免疫应答。这个为期三年的项目的具体目标是： 1)证明A-NK细胞可以用腺病毒载体转导以产生显著量的细胞因子; 2)分析细胞因子基因转导的T-LAK和A-NK细胞在肿瘤中定位和在肿瘤部位产生细胞因子的能力; 3)证明细胞因子基因转导的A-NK和T-LAK细胞的过继转移可消除肿瘤，而不会诱导全身毒性。 为了实现这一点，T-LAK和A-NK细胞将用一种或几种编码细胞因子/趋化因子的基因进行腺病毒或逆转录病毒转导，所述细胞因子/趋化因子已知通过刺激NK细胞、DC和T辅助细胞和CTL而具有抗肿瘤作用，即IL-2、IL-4、IL-12、IL-15、IL-18、IFNg、GM-CSF、RANTES和趋化因子。将在动物肿瘤模型中测试成功转导的T-LAK和A-NK细胞定位于肿瘤中并在肿瘤部位产生细胞因子的能力。将测量由肿瘤内产生的细胞因子诱导的宿主应答，例如效应细胞在肿瘤组织中的累积增加。将通过分析由注射A-NK和/或T-LAK细胞批次引起的肿瘤减少来评估该方法的治疗潜力，每种细胞能够产生一种或几种抗癌细胞因子。将阐明观察到的任何抗肿瘤作用背后的机制。 根据肿瘤寻求A-NK和T-LAK细胞产生的细胞因子，我们预计我们可以操纵宿主免疫系统，以引发强的先天性和适应性抗肿瘤反应，而不会诱导毒副作用。在此背景下，有可能显著提高肿瘤靶向细胞因子基因治疗癌症的疗效。