Project 3: Increasing the potency and accessibility of EBVSTs for the treatment of Lymphoma

项目摘要/摘要 项目3的主要目标是设计和实施有效、低毒的EBV特异性T细胞的新策略 EBV阳性淋巴瘤的EBVST细胞疗法；约占所有人类淋巴瘤的30%。淋巴瘤 环境缺乏维持EBVSTs抗肿瘤活性所需的积极信号，相反 表达产生EBVST无能的抑制信号。我们的结构性白介素7受体(C7R)不仅 提供内在的细胞因子信号，但激活抗凋亡程序，从而提供对肿瘤的抵抗 导致T细胞功能障碍。C7R导致GD2CAR T细胞的扩张，类似于淋巴耗竭 神经母细胞瘤患者的化疗(LD)与LD不同，不会导致细胞因子释放综合征 (CRS)。EBVSTs中C7R的表达可更快清除自体EBV转化的B细胞 NSG异种移植模型中的肿瘤，在目标1中，我们将检验C7R将增强 淋巴瘤患者中EBVST的扩张和持续，将提供对 免疫抑制肿瘤微环境，并提供一种无毒的替代LD。 EBV-再激活可使注入的自体EBVSTs呈指数级扩张，并促进肿瘤清除 EBV+淋巴瘤患者。在目标2中，我们将对EBV潜伏期反转剂帕诺比妥进行临床评估 (PBST)，以增强C7R-EBVST的扩展和功能。PBST还具有直接的抗淋巴瘤活性。 通过下调STAT5和增加促凋亡caspase的表达。这些抑制活性 将在EBVSTs中被上调STAT5的C7R所抵消。因此，我们的第二个假设(目标2)是 PBST将同时增强C7R-EBVST的抗原刺激作用，同时选择性增加 肿瘤细胞对C7R-EBVST杀伤的敏感性。 EBV+淋巴瘤的理想治疗方法是现成的、部分匹配的C7R-EBVSTs，这些细胞 有效的，立即可用的，并且相对便宜的。然而，移植物抗宿主病(GVHD)和移植物 同种异体反应性T细胞引起的排斥反应降低了同种异体细胞疗法的临床疗效。AIM 3将评估 CD30.CAR修改的C7R-EBVST为了解决这两个问题，EBVST在数百个月中没有产生GVHD 干细胞移植环境中的异基因接受者，由于同种异体T细胞上调CD30，他们 应该被CD30消除。事实上，银行CD30。CAR-EBVST已经安全地产生了临床 在8例CD30+淋巴瘤患者中，6例有效(3例完全)。然而，CD30.CAR-EBVST并未持续存在。 因此，目标3将检验C7R将增强CD30的假设。CAR-EBVST持久性和抗肿瘤 小鼠同种异体排斥反应模型中的活性。这三个假设将在三个具体目标中得到检验： 目的1.C7R能否增强EBV+淋巴瘤患者EBVSTs的扩张性和长期活性？ 目的2.帕诺比妥能否促进患者的EBVST扩张和抗肿瘤活性？ 目的3.CD30.CAR能否在小鼠异基因排斥模型中阻止C7R-EBVSTs的排斥反应？

PROJECT SUMMARY / ABSTRACT The broad goal of Project 3 is to devise and implement novel strategies of effective, low-toxicity EBV-specific T cell (EBVST) therapy for EBV-positive lymphomas; approximately 30% of all human lymphomas. The lymphoma environment lacks the positive signaling required to maintain the anti-tumor activities of EBVSTs and instead expresses inhibitory signals producing EBVST anergy. Our constitutive interleukin 7 receptor (C7R) not only provides intrinsic cytokine signaling but activates an anti-apoptotic program that provides resistance to tumor induced T-cell dysfunction. C7R produced expansion of GD2.CAR T-cells that was similar to lymphodepleting chemotherapy (LD) in patients with neuroblastoma, and unlike LD, did not induce cytokine release syndrome (CRS). C7R expression in EBVSTs produced more rapid clearance of autologous EBV-transformed B-cell tumors in an NSG xenograft model and in Aim 1, we will test the hypothesis that C7R will enhance the expansion and persistence of EBVSTs in patients with lymphoma, will provide resistance to the immunosuppressive tumor microenvironment and provide a non-toxic alternative to LD. EBV-reactivation produces exponential expansion of infused autologous EBVSTs and tumor clearance in patients with EBV+ lymphoma. In Aim 2 we will clinically evaluate an EBV latency reversal agent, panobinostat (PBST) to enhance the expansion and function of C7R-EBVSTs. PBST also has direct anti-lymphoma activity by downregulating STAT5 and increasing the expression of pro-apoptotic caspases. These inhibitory activities will be counteracted in EBVSTs by the C7R that upregulates STAT5. Thus, our second hypothesis (Aim 2) is that PBST will simultaneously increase antigen stimulation of C7R-EBVST while selectively increasing susceptibility of the tumor cells to C7R-EBVST killing. The ideal therapy for EBV+ lymphoma would be off-the-shelf, partially-HLA matched C7R-EBVSTs that are potent, immediately available, and relatively inexpensive. However, graft versus host disease (GVHD) and graft rejection caused by alloreactive T-cells curtail the clinical efficacy of allogeneic cell therapies. Aim 3 will evaluate CD30.CAR-modified C7R-EBVSTs to resolve both problems, EBVSTs have not produced GVHD in hundreds of allogeneic recipients in the stem cell transplant setting, and since alloactivated T-cells upregulate CD30, they should be eliminated by the CD30.CAR. Indeed banked CD30.CAR-EBVSTs have safely produced clinical responses (3 complete), in 6 of 8 patients with CD30+lymphoma. However, CD30.CAR-EBVSTs did not persist. Hence Aim 3 will test the hypothesis that C7R will enhance CD30.CAR-EBVST persistence and anti-tumor activity in a murine allo-rejection model. These 3 hypotheses will be tested in the three specific Aims: Aim 1. Can C7R enhance expansion, and long-term activity of EBVSTs in patients with EBV+ lymphoma? Aim 2. Can panobinostat boost EBVST expansion and anti-tumor activity in patients? Aim 3. Can CD30.CAR prevent rejection of C7R-EBVSTs in a murine allo-rejection model?