Adoptive Cellular Therapy of T Cell Lymphoma

项目摘要 外周T细胞淋巴瘤（PTCL）是一组高度异质性的肿瘤，占10-30%， 所有非霍奇金淋巴瘤。其中，血管免疫母细胞性T细胞淋巴瘤（AITL）、PTCL NOS（未 另外指明），和塞扎里综合征（SS）特别具有侵袭性，5年生存率<30%。 因此，积极的化疗，靶向药物和干细胞移植的疗效有限。事实上， 多达26-59%的患者由于早期复发或原发性难治性甚至从未达到移植。 目前迫切需要能够治愈或至少使疾病消退以允许移植的新疗法。AITL和A PTCL NOS亚群共享T滤泡辅助细胞表型，沿着几乎一半的SS，它们都 表达趋化因子受体CXCR 5。对于AITL，CXCR 5信号传导可能有助于其发病机制， 使其成为一个特别好的目标。靶向CD 19的嵌合抗原受体（CAR）T细胞已经显示出 在治疗B细胞白血病和淋巴瘤中具有显著的疗效。在初步工作中，我们开发了一个 靶向CXCR 5的新型嵌合抗原受体（CAR）。重要的是，细胞毒性CD 8 + T细胞用于 表达CXCR 5 CAR的人本身对CXCR 5是阴性的，从而避免了自相残杀。我们的中央 假设CXCR 5 CART将在大的肿瘤中高度有效地治疗恶性T细胞克隆， 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CXCR 5 CAR的疗效。 在体外、离体和体内的多种PTCL模型上。根据最近的观察， 优化提高CAR活性并减少副作用，在目标2中，我们将比较CXCR 5的功效， 具有一系列亲和力的汽车。在初步工作中，我们已经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CXCR 5结合剂。在目标3中，我们将测试以下物质的协同细胞毒性： 同时用维奈托克激活细胞凋亡的内在途径，沿着外在途径 通过CXCR 5 CAR的凋亡。为了最大限度地发挥协同作用并保护CAR-T细胞免受维奈托克的侵害，我们将 用抗凋亡BCL-2家族成员额外强化CXCR 5 CAR T细胞。以避免潜在 为了避免在T细胞中表达抗凋亡基因的并发症，它将以瞬时方式作为抗凋亡基因被引入。 mRNA分子。mRNA重编程具有许多优于DNA的优点，包括，除其他外， 在单个快速步骤中引入多个基因并将细胞毒性仔细校准到所需水平的能力 次给药结束本研究整合了药理学、遗传学、免疫学和细胞凋亡学的成功方法 开发一种新的治疗方法，可以通过改善 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.