Intratumoral genetic therapy for lung cancer

描述（由申请人提供）： 项目摘要/摘要肺癌的瘤内基因治疗我们集中研究通过对肿瘤部位的免疫环境进行遗传和细胞操作来增强非小细胞肺癌（NSCLC）的全身免疫应答。由于肺癌细胞本身不能提呈抗原，因此需要有效的专业抗原提呈细胞。我们的研究小组是第一个报告肿瘤内注射树突状细胞（DC）导致全身抗肿瘤免疫反应，导致临床前肺癌模型中转移性疾病的限制。通过改变原发部位的肿瘤微环境，我们的研究重点是恢复肿瘤抗原呈递和抗肿瘤效应物活性，从而诱导全身抗肿瘤免疫应答并减少转移性疾病。在2000年，我们首次报道了C-C基序配体-21（CCL 21，以前也称为次级淋巴组织趋化因子，exodus-2或6Ckine）在NSCLC的临床前模型中导致有效的免疫依赖性抗肿瘤应答。CCL 21是一种淋巴趋化因子，主要由淋巴结和派伊尔集合淋巴结中的高内皮微静脉以及脾和阑尾中的淋巴管、基质细胞组成性表达。CCL 21与趋化因子受体CCR 7结合，并且是成熟DC、初始和记忆T细胞的化学引诱物。我们随后的研究表明，体外增殖的自体DC是将CCL 21引入肿瘤部位的最有效载体。因此，我们创建了DC基因修饰以过表达趋化因子CCL 21（AdCCL 21-DC）用于瘤内注射的模型。根据我们在实验室的发现，我们已经将这项工作转化为晚期肺癌患者的临床评估。在成功地对这一概念进行竞争性同行评审后，NCI快速获得研究药物计划完成了我们表达CCL 21的腺病毒载体（Ad-CCL 21）的cGMP生产。利用这种载体，我们已经证明，可以从冷冻保存的外周血单核细胞（PBMC）中产生活的和生物活性的临床级CCL 21基因修饰的DC。这使我们能够在一小群无法手术的NSCLC患者中进行重组咨询委员会（RAC）和FDA批准的试点和可行性临床试验。这些患者正在接受腺病毒CCL 21基因修饰的DC（AdCCL 21-DC）的肿瘤内注射，并监测毒性和特异性免疫应答，如初步数据部分所示。我们现在提出一项正式的I期试验，在VA大洛杉矶医疗中心和UCLA罗纳德里根医疗中心招募21名患者，评估安全性，抗肿瘤反应和免疫反应性。在晚期NSCLC患者中的该I期试验的背景下，我们将确定肿瘤内施用的AdCCL 21-DC的安全性和最大耐受剂量。此外，将通过评估瘤内AdCCL 21-DC给药后的抗肿瘤免疫反应、肿瘤免疫组织化学、T调节细胞的变化和血浆细胞因子谱来评估局部和全身生物活性。最后，将评估反映在肿瘤负荷降低中的临床活性。这是一种全新的肺癌临床免疫干预方法;这是人类第一次以任何形式接受CCL 21，也是第一次评估肺癌肿瘤内注射DC的临床试验。

DESCRIPTION (provided by applicant): Project Summary/Abstract Intratumoral genetic therapy for lung cancer We have focused investigations on augmenting systemic immune responses in non-small cell lung cancer (NSCLC) by genetic and cellular manipulation of the immune environment at the tumor site. Because lung cancer cells themselves cannot present antigen, effective professional antigen presenting cells are required. Our research group was the first to report that the intratumoral injection of dendritic cells (DC) led to systemic antitumor immune responses resulting in limitation of metastatic disease in pre-clinical lung cancer models. By altering the tumor microenvironment at the primary site, our investigations focus on restoring tumor antigen presentation and antitumor effector activities that can induce systemic antitumor immune responses and reduce metastatic disease. In 2000, we reported for the first time that C-C motif ligand-21 (CCL21, also previously referred to as secondary lymphoid tissue chemokine, exodus-2 or 6Ckine) led to potent immune- dependent antitumor responses in preclinical models of NSCLC. CCL21 is a lymphoid chemokine that is predominantly and constitutively expressed by high endothelial venules in lymph nodes and Peyer's patches and by lymphatic vessels, stromal cells in the spleen and appendix. CCL21 binds to the chemokine receptor CCR7 and is chemoattractant for mature DCs, naive and memory T cells. Our subsequent studies revealed that in vitro propagated autologous DC served as the most effective vehicle for introduction of CCL21 to the tumor site. We therefore created models in which DC were gene modified to over express the chemokine CCL21 (AdCCL21-DC) for intratumoral injection. Based on our findings in the laboratory, we have translated this work for clinical evaluation for late stage lung cancer patients. Following successful competitive peer review of this concept, the NCI Rapid Access to Investigational Drug program completed cGMP production of our adenoviral vector expressing CCL21 (Ad-CCL21). Utilizing this vector we have demonstrated that viable and biologically active clinical grade CCL21 gene-modified DC can be generated from cryopreserved peripheral blood mononuclear cells (PBMC). This has allowed us to conduct a Recombinant Advisory Committee (RAC) and FDA-approved pilot and feasibility clinical trial in a small group of patients with inoperable NSCLC. These patients are receiving intratumoral injection of adenoviral CCL21 gene modified DC (AdCCL21-DC) and are monitored for toxicity and specific immune responses as presented in the preliminary data section. We now propose a formal phase I trial to enroll 21 patients at the VA Greater Los Angeles Healthcare Center and UCLA Ronald Reagan Medical Center in an assessment of safety, anti-tumor response and immune reactivity. In the context of this phase I trial in patients with advanced NSCLC we will determine the safety and maximum tolerated dose of intratumorally administered AdCCL21-DC. In addition, the local and systemic biological activity will be evaluated by assessment of anti-tumor immune responses, tumor immunohistochemistry, alterations in T regulatory cells and plasma cytokine profiles following intratumoral AdCCL21-DC administration. Finally, the clinical activity as reflected in reduction in tumor burden will be assessed. This is an entirely new approach to clinical immune-based intervention in lung cancer; this is the first time that humans are receiving CCL21 in any form and the first clinical trial to assess intratumoral injection of DC in lung cancer.