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

项目概要/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细胞治疗的新策略。 细胞（EBVST）治疗EBV阳性淋巴瘤;约占所有人类淋巴瘤的30%。淋巴瘤 环境缺乏维持EBVST抗肿瘤活性所需的正信号传导， 表达产生EBVST无反应性的抑制信号。我们的组成型白细胞介素7受体（C7 R）不仅 提供内在的细胞因子信号，但激活抗凋亡程序，提供对肿瘤的抗性 诱发T细胞功能障碍C7 R产生与淋巴细胞耗竭类似的GD2.CAR T细胞扩增。 在神经母细胞瘤患者中，与LD不同的是，LD不会诱导细胞因子释放综合征 （CRS）。EBVSTs中C7 R的表达使自体EBV转化B细胞的清除更快 在NSG异种移植物模型中的肿瘤和目标1中，我们将检验C7 R将增强NSG异种移植物模型中的肿瘤的细胞增殖的假设。 在淋巴瘤患者中，EBVST的扩展和持续，将提供对 免疫抑制肿瘤微环境，并提供LD无毒替代物。 EBV再活化导致输注的自体EBVST的指数扩增和肿瘤清除， EBV+淋巴瘤患者。在目标2中，我们将临床评估EBV潜伏期逆转剂帕比司他 （PBST）以增强C7 R-EBVST的扩增和功能。PBST还具有直接的抗淋巴瘤活性 通过下调STAT 5和增加促凋亡caspase的表达。这些抑制活性 在EBVST中会被上调STAT 5的C7 R抵消。因此，我们的第二个假设（目标2）是 PBST将同时增加C7 R-EBVST的抗原刺激， 肿瘤细胞对C7 R-EBVST杀伤的敏感性。 EBV+淋巴瘤的理想疗法是现成的HLA部分匹配的C7 R-EBVST， 有效的，立即可用的，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细胞引起的排斥减少了同种异体细胞疗法的临床功效。目标3将评估 CAR修饰的C7 R-EBVST解决了这两个问题，EBVST在数百例移植物中没有产生GVHD。 在干细胞移植环境中的同种异体受体，由于同种异体活化的T细胞上调CD 30， 应该通过CD30.CAR消除。事实上，库存的CD 30。CAR-EBVST已安全地产生临床 8例CD 30+淋巴瘤患者中有6例缓解（3例完全缓解）。然而，CD 30. CAR-EBVST不持续存在。 因此，目标3将检验C7 R将增强CD 30、CAR-EBVST持久性和抗肿瘤活性的假设。 在小鼠同种异体排斥模型中的活性。将在三个特定目标中检验这三个假设： 目标1。C7 R能否增强EBV+淋巴瘤患者中EBVSTs的扩增和长期活性？ 目标二。帕比司他能否促进患者的EBVST扩增和抗肿瘤活性？ 目标3。在小鼠同种异体排斥模型中，CD30.CAR能否预防C7 R-EBVSTs的排斥？

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?