Cellular Immunotherapy of Ovarian Cancer

项目摘要 大多数复发和晚期卵巢癌(OC)患者的治疗非常有限。 选择。检查点封锁在不到15%的患者中显示出客观反应。因此， 发展更有效的免疫治疗方法，如嵌合抗原受体(CAR)T细胞 (CAR-TS)对这些患者来说是关键的。我们已经确定B7-H3是OC中CART的有效靶点。我们有 获得了B7-H3.CAR-ts，并成功地在包括OC模型在内的实体瘤模型中进行了测试。此外, 自从B7-H3CAR以来，我们已经与mB7-H3发生了交叉反应，我们进行了有效性和安全性研究 在免疫活性强的小鼠中表现出无毒的抗肿瘤活性。已确定并验证B7-H3为 作为OC的目标，在本应用中，我们旨在克服肿瘤微环境(TME) 免疫抑制在OC中充分发挥CAR技术的潜力。对OC中的TME进行了表征 通过促进血管生成和形成免疫抑制细胞的细胞网络。尤其是肿瘤 伴发巨噬细胞(TAMs)和髓系抑制细胞(MDSCs)在TME中含量丰富 OC，抑制效应性T细胞。与T细胞相比，NKT具有与T细胞共定位的先天特性 并通过其不变的TCR(ITCR)以CD1d依赖的方式利用TAMs的裂解作用。我们 已经发表和生成了更多的初步数据，表明会表达汽车的人类NKT是双重的 通过CAR特异性靶向肿瘤细胞，通过天然iTCR特异性靶向TAMS。此外，我们还产生了 初步数据显示，人类NKT也可以被改造成释放IL-12，这是一种已知的细胞因子 对MDSC重新编程。我们假设NKT基因工程表达B7-H3CAR和IL-12将克服 实体肿瘤过继免疫治疗的关键挑战：效应细胞在肿瘤部位的定位 通过B7-H3CAR选择性杀伤肿瘤细胞，通过CD1d结合消除肿瘤保护性TAMs ITCR，并通过IL-12对MDSCs重新编程。我们新的初步数据还显示，IL-12具有很强的 可能通过一种新的机制增强NKT中CD62L相关的干样程序，相关的唯一 “维生素D签名”。因此，我们假设人类NKT可能具有一种不同于以前的内在可塑性 IL-12可能通过维生素D途径将NKT重新编程为更不成熟的表型。三 提出了具体目标： 目的1：评价NKT的B7-H3CAR和IL-12工程及天然iTCR在靶向方面的协同作用 OC细胞和形成免疫活性小鼠模型中的TME。 目的2：机械评估NKT表达的IL-12如何促进NKT持续时间更长 收养转移。 目的：评价工程化人NKT在人免疫肿瘤(HIT)小鼠体内的抗肿瘤活性。

Project Abstract The majority of women with relapsed and advanced ovarian cancer (OC) have then very limited therapeutic options. Checkpoint blockade has shown objective responses in less than 15% in patients. Therefore, the development of more potent immunotherapy approaches such as chimeric antigen receptor (CAR) T cells (CAR-Ts) is critical in these patients. We have identified B7-H3 as a valid target for CAR-Ts in OC. We have generated B7-H3.CAR-Ts and successfully tested them in solid tumor models including OC models. In addition, since the B7-H3.CAR we have developed cross-reacts with mB7-H3, we conducted efficacy and safety studies in immunocompetent mice showing antitumor activity without toxicity. Having identified and validated B7-H3 as a target for OC, in this application we aim at overcoming the tumor microenvironment (TME) immunosuppression in OC to fully exploit the potential of the CAR technology. The TME in OC is characterized by a cellular network that promotes angiogenesis and shapes immunosuppressive cells. In particular, tumor associated macrophages (TAMs) and myeloid-derived suppressive cells (MDSCs) are abundant in the TME of OC, and inhibit effector T cells. As compared to T cells, NKTs possess the innate property to co-localize with TAMs and to exploit lytic effects on TAMs in a CD1d-dependent manner via their invariant TCR (iTCR). We have published and generated additional preliminary data showing that CAR-expressing human NKTs are dual specific targeting both tumor cells via CAR and TAMs via native iTCR. Furthermore, we have generated preliminary data showing that human NKTs can also be engineered to release IL-12, a cytokine known to reprogram MDSCs. We hypothesize that NKTs engineered to express the B7-H3.CAR and IL-12 will overcome critical challenges of adoptive immunotherapy of solid tumors: effector cell localization to the tumor site, selective killing of tumor cells via B7-H3.CAR, elimination of tumor-protective TAMs via CD1d engagement by the iTCR, and reprogramming of MDSCs via IL-12. Our new preliminary data also revealed that IL-12 potently enhances CD62L-associated stem-like program in NKTs likely via a novel mechanism, associated with unique “vitamin D signature”. We thus hypothesize that human NKTs may have an intrinsic plasticity not previously appreciated, and that IL-12 may reprogram NKTs to a more immature phenotype via vitamin D pathway. Three specific Aims are proposed: Aim 1: To evaluate whether B7-H3.CAR and IL-12 engineering of NKTs and native iTCR cooperate in targeting OC cells and shaping the TME in an immunocompetent murine model. Aim 2: To mechanistically assess how IL-12 expressed by NKTs promotes NKTs with longer persistence upon adoptive transfer. Aim 3: To evaluate the antitumor activity of engineered human NKTs in Human-Immune Tumor (HIT) mice.