Vaccination to Enhance T cell Immunotherapy

项目1(C.Rooney和L.Wang)的长期目标是提高 使用两种新方法过继转移肿瘤特异性T细胞。(1)以下第三人成立为法团- GD2嵌合抗原受体(CAR)的产生 CD28和OX40被假设为增强和维持T细胞在肿瘤内的反应，而 提供对肿瘤微环境中阐述的抑制配体的抵抗力。(Ii)及 将GD2.CAR基因移植到特异性T细胞可促进瘤外增殖 对于水痘-带状疱疹病毒(VZV)，存在一种有效的减毒活疫苗 这通过其TCR增加了VZV特异性T细胞在体内的增殖。这些宽泛的概念将 在GD2.CAR移植VZV特异性T细胞治疗慢性粒细胞白血病的临床试验中被探索 晚期GD2阳性肉瘤(A，I，S 1，2)。因为这将是一项“第一人”研究， 转导的T细胞还将结合基于caspase-9二聚化的可诱导安全开关 分子，它在最近的一项治疗移植物抗宿主疾病的临床试验中得到了验证。不幸的是， 最年轻的儿童肉瘤患者将是VZV阴性，这使得产生VZV特异性变得困难 并证明这些血清阴性患者接种减毒活病毒是合理的。 因此，Aim 3将在临床前模型中评估一种针对GD2的细胞疫苗，该疫苗应该能够提供 通过汽车进行肿瘤外刺激。这一努力将依赖于JF神经母细胞瘤(NB)细胞系，该细胞系具有 作为一种针对新城疫的细胞疫苗进行了广泛的测试。GD2在JF细胞中强表达，并可 由JFNB细胞直接和局部抗原间接提呈给GD2的CAR阳性T细胞- 呈现细胞。对JFNB细胞进行基因改造，以表达细胞因子，如ILI5和GM-CSF， 促进T细胞增殖和募集和激活树突状细胞应促进瘤外增殖 以类似于VZV刺激天然T细胞受体的方式表达CAR阳性T细胞。部分地 因此，我们的项目使用了项目3中开发的概念，并提供了潜在的方法来增加 项目2和3(iC9安全基因)和项目4(针对转化生长因子β的DNR)的安全性和有效性。 相关性(请参阅说明)： T细胞治疗晚期肉瘤和其他高危实体瘤的研究进展 由于靶向肿瘤抗原的弱表达或缺失以及表现不佳而减慢 输注T细胞。项目1试图通过结合改进的抗原受体来解决这个问题 通过新的疫苗接种策略确保内外充分的T细胞刺激能力 肿瘤部位。

The long-term goal of Project 1 (C. Rooney and L. Wang) is to improve the expansion and persistence of adoptively transferred tumor-specific T cells using two novel approaches. (1) Incorporation of a third- generation chimeric antigen receptor (CAR) for GD2 containing intracellular signaling domains for CD28 and OX40 is hypothesized to enhance and sustain T- cell responses intratumorally, while providing resistance to inhibitory ligands elaborated within the tumor microenvironment. (ii) An extratumoral proliferative boost should be attained by engrafting the GD2.CAR onto T cells specific for the varicella-zoster virus (VZV), for which there exists a potent live-attenuated booster vaccine that increases the in vivo proliferation of VZV-specific T cells via their TCRs. These broad concepts will be explored in a clinical trial of GD2.CAR-engrafted VZV-specific T cells for the treatment of patients with advanced GD2-positive sarcomas ( A i m s 1 and 2). Because this will be a "first-in-man" study, the transduced T cells will also incorporate an inducible safety switch based on dimerization of the caspase-9 molecule, which was validated in a recent clinical trial against graft-vs.-host disease. Unfortunately, the youngest pediatric sarcoma patients will be VZV-negative, making it difficult to generate VZV-specific autologous T cells and to justify vaccination of these seronegative patients with a live-attenuated virus. Hence, Aim 3 will evaluate in a preclinical model a cellular vaccine against GD2 that should provide extratumoral stimulation via the CAR. This effort will rely on the JF neuroblastoma (NB) cell line, which has been extensively tested as a cellular vaccine for NB. GD2 is strongly expressed by JF cells and can be presented to GD2,CAR-positive T cells both directly by the JFNB cells and indirectly by local antigen- presenting cells. Genetic modification of JFNB cells to express cytokines, such as ILI 5 and GM-CSF, that enhance T-cell proliferation and recruit and activate dendritic cells should promote extratumoral proliferation of the CAR-positive T cells in ananalogous way to stimulation of the native T cell receptor by VZV. In part therefore our project uses concepts developed in Project 3 and provides potential ways of increasing the safety and efficacy of projects 2 and 3 (the iC9 safety gene) and Project 4 (the DNR for TGFbeta). RELEVANCE (See instructions): Progress in the development of T-cell therapy for advanced sarcomas and other high-risk solid tumors has been slowed by the weak expression or absence of targetable tumor antigens and poor performance of infused T cells . Project 1 seeks to address this problem by combining improved antigen receptor capabilities with a novel vaccination strategy to ensure adequate T-cell stimulation, both within and outside the tumor site.