SYNERGIZING PRO-APOPTOTIC AND CAR-T CELL IMMUNOTHERAPY FOR MANTLE CELL LYMPHOMA

 描述（由申请人提供）：套细胞淋巴瘤（MCL）是一种高度侵袭性的B细胞淋巴瘤，对常规化疗耐药。虽然定义的特征t（11;14）易位，导致细胞周期增殖基因cyclin D的过度表达，各种遗传畸变已被确定在MCL，并可能有助于其发病机制和耐药性。特别令人感兴趣的是抗凋亡BCL-2的频繁增加以及促凋亡BIM的缺失。因此，该提议将探索同时激活MCL中的两个促凋亡途径的协同细胞毒性。首先，以BCL-2同源3（BH 3）死亡结构域为模型的小分子，称为BH 3模拟物，将诱导细胞凋亡的内在途径。其次，用靶向CD 19的嵌合抗原受体（汽车）重编程的细胞毒性T细胞将诱导细胞凋亡的外源性途径。尽管CAR-T细胞对先前无反应的B细胞恶性肿瘤显示出令人印象深刻的疗效，但它们易受其他细胞毒性化疗和凋亡刺激的影响-限制了它们在联合治疗中的疗效。因此，为了最大化协同作用，我们将测试各种基因在BH 3模拟物存在下赋予CAR-T细胞存活优势的能力。目的1将测试各种抗凋亡BCL-2家族成员保护CAR-T细胞免受BH 3模拟物诱导的细胞死亡的使用。我们假设，未被选择的BH 3模拟物靶向的抗凋亡蛋白将对CAR-T细胞提供保护。目的2将测试在BH 3模拟物存在下使用自刺激细胞因子-细胞因子受体促进T细胞存活。目的3将在体内MCL异种移植模型中探索由抗凋亡因子和细胞因子因子两者强化的BH 3模拟物和CAR-T细胞的组合。为了避免在T细胞中表达抗凋亡和增殖基因的潜在并发症，它们将以瞬时方式作为mRNA分子引入。mRNA重编程具有许多优于DNA的优点，其中包括在单个快速步骤中引入多个基因的能力。通过整合来自化学、免疫学和细胞凋亡的成功方法，我们能够以一种能够通过其他靶向治疗模式实现更稳健、校准的细胞毒性的方式显著改善CAR-T细胞技术。

 DESCRIPTION (provided by applicant): Mantle Cell Lymphoma (MCL) is a highly aggressive B-cell lymphoma resistant to conventional chemotherapy. Although defined by the characteristic t(11;14) translocation that results in overexpression of the cell cycle proliferation gene cyclin D, a variety of genetic aberrations have been identified in MCL and may contribute to its pathogenesis and chemoresistance. Of particular interest are frequent gains of anti-apoptotic BCL-2 as well as deletions of pro-apoptotic BIM. Therefore, this proposal will explore the synergistic cytotoxicity of simultaneously activating two pro-apoptotic pathways in MCL. First, small molecules modeled after the BCL-2 Homology 3 (BH3) death domain, known as BH3- mimetics, will induce the intrinsic pathway of apoptosis. Second, cytotoxic T cells reprogrammed with chimeric antigen receptors (CARs) that target CD19 will induce the extrinsic pathway of apoptosis. Although CAR-T cells have displayed impressive efficacy toward previously unresponsive B cell malignancies, they are vulnerable to other cytotoxic chemotherapies and apoptotic stimuli - limiting their efficacy in combination therapies. Therefore, in order to maximize synergy, we will test the ability of various genes to impart a survival advantage to the CAR-T cells while in the presence of BH3 mimetics. Aim 1 will test the use of various anti-apoptotic BCL-2 family members to protect CAR-T cells from BH3 mimetic-induced cell death. We hypothesize that anti-apoptotic proteins not targeted by select BH3 mimetics will confer protection to the CAR-T cells. Aim 2 will test the use of a self-stimulating cytokine- cytokine receptor to promote T cell survival in the presence of BH3 mimetics. Aim 3 will explore the combination of BH3 mimetics and CAR-T cells, fortified by both the anti-apoptotic and cytokine factors, in an MCL xenograft model in vivo. To avoid potential complications of expressing anti-apoptotic and proliferative genes in a T cell, they will be introduced in a transient fashion as mRNA molecules. mRNA reprogramming has numerous advantages over DNA including, among others, the ability to introduce multiple genes in a single rapid step. By integrating successful approaches from chemistry, immunology and apoptosis we are well positioned to significantly improve CAR-T cell technology in a manner that enables a more robust, calibrated cytotoxicity with additional modes of targeted therapeutics.