Adoptive Cellular Therapy of T Cell Lymphoma

项目总结 外周T细胞淋巴瘤(PTCL)是一组高度异质性的肿瘤，占10%-30% 在所有非霍奇金淋巴瘤中。其中，血管免疫母细胞T细胞淋巴瘤(AITL)、PTCL-NOS(非 另外，Sezary综合征(SS)尤其具有侵袭性，5年存活率达30%。 因此，积极的化疗、靶向药物和干细胞移植的效果有限。事实上, 多达26-59%的患者由于早期复发或原发难治性疾病而从未进行过移植。 迫切需要能够治愈或至少缓解疾病以允许移植的新疗法。AITL和a PTCL NOS的亚群具有T滤泡辅助细胞表型，并且与近一半的SS一起，它们都 表达趋化因子受体CXCR5。对于AITL，CXCR5信号可能参与了其发病机制。 使其成为一个特别好的目标。针对CD19的嵌合抗原受体(CAR)T细胞已显示 治疗B细胞白血病和淋巴瘤疗效显著。在前期工作中，我们开发了一种 针对CXCR5的新型嵌合抗原受体(CAR)。重要的是，细胞毒性CD8+T细胞用于 快递的CXCR5车，本身都是CXCR5阴性，从而避免了自相残杀。我们的中央 假设CXCR5 CART将对治疗大量的恶性T细胞克隆非常有效 PTCL患者亚组。为了验证这一假设，在目标1中，我们将评估我们的CXCR5汽车的效率 在体外、体外和体内建立多种PTCL模型。基于最近的观察，亲和力 优化提高CAR活跃度并减少副作用，在目标2中我们将比较CXCR5的疗效 具有一系列亲和力的汽车。在前期工作中，我们开发了一个噬菌体筛选平台，该平台具有 鉴定了具有不同亲和力的CXCR5粘结剂。在目标3中，我们将测试其协同细胞毒性。 静脉注射同时激活细胞凋亡的内源性途径和外源性途径 CXCR5 CAR诱导的细胞凋亡。为了最大限度地发挥协同作用并保护CAR-T细胞免受万诺克拉病毒的侵袭，我们将 此外，CXCR5 CAR T细胞还含有抗凋亡的bcl2家族成员。为了避免潜在的 在T细胞中表达抗凋亡基因的并发症，它将作为一种瞬时方式引入 信使核糖核酸分子与DNA相比，信使核糖核酸重新编程有许多优势，其中包括 能够在单个快速步骤中引入多个基因，并仔细校准所需的细胞毒性 剂量。这项研究综合了药理学、遗传学、免疫学和细胞凋亡的成功方法。 开发一种新的治疗方法，通过改进的 成为移植或最终治愈的桥梁。

PROJECT SUMMARY Peripheral T cell lymphomas (PTCL) are a highly heterogeneous group of neoplasms that account for 10-30% of all Non-Hodgkin Lymphomas. Of these, angioimmunoblastic T cell lymphoma (AITL), PTCL NOS (not otherwise specified), and Sezary syndrome (SS) are particularly aggressive with 5-year survivals of <30%. Thus, aggressive chemotherapy, targeted agents and stem cell transplantation have limited efficacy. In fact, as many as 26-59% of patients never even reach transplantation due to early relapse or primary refractoriness. Novel therapies that can cure, or at least subside disease to allow transplant, are urgently needed. AITL and a subset of PTCL NOS share a T-follicular helper cell phenotype, and along with nearly half of SS, they all express the chemokine receptor CXCR5. For AITL, CXCR5 signaling may contribute to its pathogenesis, making it a particularly good target. Chimeric Antigen Receptor (CAR) T cells targeting CD19 have shown remarkable efficacy in treating B cell leukemias and lymphomas. In preliminary work, we have developed a novel Chimeric Antigen Receptor (CAR) that targets CXCR5. Importantly, cytotoxic CD8+ T cells used to express the CXCR5 CAR, are negative for CXCR5 themselves, thus avoiding fratricide. Our central hypothesis is that CXCR5 CART will be highly effective for the treatment of malignant T cell clones in a large subset of patients with PTCL. To test this hypothesis, in aim 1 we will assess the efficacy of our CXCR5 CAR on multiple models of PTCL in vitro, ex vivo, and in vivo. Building upon recent observations that affinity optimization improves CAR activity and reduces side effects, in aim 2 we will compare the efficacy of CXCR5 CARs with a range of affinities. In preliminary work, we have developed a phage screening platform that has identified CXCR5 binders with different affinities. In aim 3 we will test the synergistic cytotoxicity of simultaneously activating the intrinsic pathway of apoptosis with Venetoclax, along with the extrinsic pathway of apoptosis by the CXCR5 CAR. To maximize synergy and protect the CAR-T cells from Venetoclax, we will additionally fortify the CXCR5 CAR T cells with an anti-apoptotic BCL-2 family member. To avoid potential complications of expressing an anti-apoptotic gene in a T cell, it will be introduced in a transient fashion as an mRNA molecule. mRNA reprogramming has numerous advantages over DNA including, among others, the ability to introduce multiple genes in a single rapid step and to carefully calibrate cytotoxicity to the required dose. This study integrates successful approaches from pharmacology, genetics, immunology and apoptosis to develop a novel therapy that could meaningfully improve patient outcomes, through either an improved bridge to transplant or an eventual cure.