Third-party “off the shelf” mature or precursor CAR T cells to prevent or treat malignant relapse after allo HCT

第三方“现成的”成熟或前体 CAR T 细胞，用于预防或治疗异基因 HCT 后的恶性复发

• 批准号: 9762469
• 负责人: Marcel R M van den Brink
• 金额: $ 68.32万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762469
• 关键词: Adoptive Cell Transfers; Adoptive Transfer; Allogenic; Antigen Targeting; Antigens; Antiviral Agents; Autoantigens; B-Cell Acute Lymphoblastic Leukemia; B-Lymphocytes; CD19 gene; CRISPR/Cas technology; Cause of Death; Cells; Clinical; Clinical Research; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Disease remission; Engineering; Failure; Genetic Engineering; Graft Rejection; Hematologic Neoplasms; Hematology; Hematopoietic; Hematopoietic Neoplasms; Human; Immunity; In Vitro; Infusion procedures; Leucine Zippers; Lymphocyte; Lymphoma; Malignant - descriptor; Malignant Neoplasms; Mature T-Lymphocyte; Mediating; Mus; Patients; Prevention; Production; Recovery; Recurrent disease; Refractory; Regulation; Relapse; Risk; Series; Signal Transduction; System; T cell therapy; T-Cell Receptor; T-Lymphocyte; Techniques; Thymus Gland; Time; Tissues; Translations; Transplant Recipients; Transplantation; Tumor Antigens; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; Virus Diseases; WT1 gene; base; cellular engineering; central tolerance; chimeric antigen receptor; clinical efficacy; cytokine; disorder risk; effective therapy; effector T cell; exhaustion; experience; graft failure; graft vs host disease; hematopoietic cell transplantation; immune reconstitution; in vivo; innovation; large cell Diffuse non-Hodgkin' s lymphoma; leukemia; leukemia/lymphoma; mortality; novel strategies; post-transplant; pre-clinical; preclinical study; prevent; receptor expression; transgene expression; transplant model

Abstract Relapse remains the most important cause of mortality after allogeneic hematopoietic cell transplantation (allo-HCT) and little progress has been made in the past decades. Chimeric antigen receptors targeting CD19 (CD19-CARs) redirect T cell effector functions to eliminate CD19-expressing leukemia and lymphoma cells. We found in pre-clinical studies that allogeneic donor-derived CD19-CAR T cells can have significant anti-lymphoma activity with minimal graft- versus-host disease (GVHD), an observation that was confirmed by others in clinical trials. We found that this was due to exhaustion and deletion of CAR-T cells attributable to cumulative signaling through both the alloreactive TCR and the CAR. In addition, we have demonstrated the feasibility of deletion of the endogenous TCR and expression of a CD19-CAR in T cells using CRISPR technology, which could further reduce potential alloreactivity of these cells. We have also developed an in vitro culture system to generate universal third-party T cell precursors (preT) that can be engineered to express CD19-CAR post-thymically to avoid negative selection. Upon adoptive transfer in allo-HCT recipients, these engineered preT cells can mature in the host’s thymus and exert anti-malignancy activity without GVHD. In addition to anti-malignancy potential, these engineered preT cells can also enhance immune reconstitution. Based on these findings and the clinical efficacy of adoptively transferred third party anti-viral T cells in allo-HCT recipients, we hypothesize that infusion of third-party “off-the-shelf” mature or preT cells expressing CARs and TCRs targeted against tumor-associated antigens will promote anti-malignancy activity and enhance immune reconstitution in allo-HCT recipients with minimal or no GVHD. We propose pre- clinical studies with engineered third-party mature and preT cells to prevent or treat relapse after allo-HCT. In Aim 1, we will use pre-clinical allo-HCT models to evaluate the anti-malignancy activity, GVHD potential and persistence of third-party T cells whose endogenous TCR has been deleted using CRISPR and that express (1.1) CD19-CAR, (1.2) triple-antigen-specific CARs, or (1.3) CD19-CAR/WT-1 specific TCR. In Aim 2, we will study third-party preT cells expressing (2.1) a CD19-CAR, (2.2) a CD19-CAR and cytokines, or (2.3) CD19-CAR/WT1 specific TCR. We have already developed two allo-HCT clinical trials with donor-derived CD19-CAR T cells or third-party preT cells, which together with our extensive clinical experience with CAR T cells will facilitate the translation of the proposed preclinical studies to decrease relapse in allo-HCT patients with hematologic malignancies using third party CAR T cells.

抽象的 复发仍然是同种异性造血细胞后死亡率的最重要原因 在过去的几十年中，移植（Allo-HCT）几乎没有取得进展。嵌合 靶向CD19（CD19-CARS）的抗原受体重定向T细胞效应器函数消除 表达CD19的白血病和淋巴瘤细胞。我们在临床前研究中发现了同种异体 供体衍生的CD19卡车T细胞可以具有显着的抗淋巴瘤活性，而最小的移植物 - 与宿主病（GVHD）相比，这一观察结果在临床试验中得到了证实。我们 发现这是由于耗尽和删除可归因于累积的CAR-T细胞 通过同种反应性TCR和汽车发出信号。此外，我们已经证明了 使用内源性TCR缺失的可行性和使用CD19卡车在T细胞中表达的可行性 CRISPR技术，可以进一步降低这些细胞的潜在同种异体反应性。我们有 还开发了一种体外培养系统，以产生通用的第三方T细胞前体（PRE） 可以在室内进行工程设计以避免选择性选择。之上 这些工程化的Pret细胞可以在Allo-HCT接受者中的收养转移，可以在主机中成熟 胸腺并发挥没有GVHD的抗恶性活性。除了抗恶性潜力外， 这些设计的PRET细胞也可以增强免疫重构。基于这些发现 以及适当转移的第三方抗病毒T细胞的临床效率 我们假设输注第三方“现成”成熟或表达汽车的Pret细胞 针对肿瘤相关抗原的TCR和TCR将促进抗恶性活性和 增强Allo-HCT接受者的免疫重建，而GVHD最少或没有GVHD。我们建议 使用工程设计的第三方成熟和Pret细胞进行临床研究，以防止或治疗浮雕 Allo-HCT。在AIM 1中，我们将使用临床前的Allo-HCT模型来评估反严重性 内源性TCR的第三方T细胞的活性，GVHD潜力和持久性 使用CRISPR和表达（1.1）CD19卡车，（1.2）三重抗原特异性汽车删除 （1.3）CD19-CAR/WT-1特异性TCR。在AIM 2中，我们将研究表达表达的第三方Pret细胞（2.1） CD19卡车，（2.2）CD19卡车和细胞因子，或（2.3）CD19-CAR/WT1特异性TCR。我们有 已经开发了两项使用供体衍生的CD19卡车T细胞或第三方的Allo-HCT临床试验 Pret细胞，以及我们在CAR T细胞中的广泛临床经验将促进 拟议的临床前研究的翻译，以减少Allo-HCT患者的退休 使用第三方CAR T细胞的血液系统恶性肿瘤。