Intratumoral genetic therapy for lung cancer

描述(由申请人提供)： 肺癌的肿瘤内基因治疗我们的研究重点是通过遗传和细胞操作肿瘤部位的免疫环境来增强非小细胞肺癌(NSCLC)的系统免疫反应。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。我们的研究小组首次报道，在临床前肺癌模型中，树突状细胞(DC)瘤内注射导致全身抗肿瘤免疫反应，从而限制了转移疾病的发生。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提呈和抗肿瘤效应活性，以诱导全身抗肿瘤免疫反应，减少转移疾病。2000年，我们首次报道了C-C基序配体-21(CCL21，以前也被称为次级淋巴组织趋化因子，Exodus-2或6Ckine)在NSCLC临床前模型中诱导了强大的免疫依赖性抗肿瘤反应。CCL21是一种淋巴趋化因子，主要由淋巴结和Peyer氏斑的高内皮微静脉以及脾和阑尾的淋巴管、间质细胞表达。CCL21与趋化因子受体CCR7结合，对成熟DC、幼稚T细胞和记忆性T细胞具有趋化作用。我们随后的研究表明，体外扩增的自体DC是将CCL21导入肿瘤部位的最有效的载体。因此，我们建立了DC被基因修饰以过表达趋化因子CCL21(AdCCL21-DC)以用于瘤内注射的模型。基于我们在实验室中的发现，我们已经将这项工作转化为对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审查后，NCI快速获得研究药物计划完成了我们表达CCL21的腺病毒载体(Ad-CCL21)的cGMP生产。利用该载体，我们证明了从深低温保存的外周血单个核细胞(PBMC)中可以获得具有生物活性的CCL21基因修饰的临床级DC。这使得我们能够在一小部分无法手术的非小细胞肺癌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。这些患者正在接受肿瘤内注射腺病毒CCL21基因修饰的DC(AdCCL21-DC)，并监测毒性和特异性免疫反应，如初步数据部分所示。我们现在建议进行一项正式的I期试验，在退伍军人管理局大洛杉矶医疗中心和加州大学洛杉矶分校罗纳德·里根医学中心招募21名患者进行安全性、抗肿瘤反应和免疫反应性的评估。在这项晚期非小细胞肺癌患者的I期试验中，我们将确定瘤内注射AdCCL21-DC的安全性和最大耐受量。此外，肿瘤内注射AdCCL21-DC后，将通过评估抗肿瘤免疫反应、肿瘤免疫组织化学、T调节细胞变化和血浆细胞因子谱来评估局部和系统的生物学活性。最后，将评估反映在肿瘤负担减轻上的临床活动性。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CCL21治疗，也是第一次评估肺癌瘤内注射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.