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 复发仍是继异体造血细胞之后最重要的死亡原因 嵌合体移植（allo-HCT）在过去几十年中几乎没有取得任何进展。 靶向 CD19 的抗原受体 (CD19-CAR) 重定向 T 细胞效应功能以消除 我们在临床前研究中发现表达CD19的白血病和淋巴瘤细胞。 供体来源的 CD19-CAR T 细胞只需最少的移植即可具有显着的抗淋巴瘤活性 抗宿主病（GVHD），这一观察结果在临床试验中得到了其他人的证实。 发现这是由于累积的 CAR-T 细胞耗尽和缺失所致 此外，我们还证明了通过同种异体反应性 TCR 和 CAR 的信号传导。 使用 T 细胞删除内源 TCR 并表达 CD19-CAR 的可行性 我们拥有 CRISPR 技术，可以进一步降低这些细胞的潜在同种异体反应性。 还开发了体外培养系统来生成通用第三方 T 细胞前体 (preT) 可以被设计为在胸腺后表达 CD19-CAR 以避免负选择。 在同种异体 HCT 受体中过继转移后，这些工程前 T 细胞可以在宿主体内成熟 胸腺并在没有 GVHD 的情况下发挥抗恶性肿瘤活性。 基于这些发现，这些工程前 T 细胞还可以增强免疫重建。 以及过继转移的第三方抗病毒 T 细胞在同种异体 HCT 受者中的临床疗效， 我们寻求输注表达 CAR 的第三方“现成”成熟细胞或 preT 细胞 针对肿瘤相关抗原的 TCR 将促进抗恶性肿瘤活性 增强异基因 HCT 受者的免疫重建，GVHD 极少或无 GVHD。 使用工程化的第三方成熟细胞和前 T 细胞进行临床研究，以预防或治疗术后复发 在目标1中，我们将使用临床前allo-HCT模型来评估抗恶性肿瘤作用。 内源性 TCR 已被检测的第三方 T 细胞的活性、GVHD 潜​​力和持久性 使用 CRISPR 删除并表达 (1.1) CD19-CAR、(1.2) 三抗原特异性 CAR，或 (1.3) CD19-CAR/WT-1 特异性 TCR 在目标 2 中，我们将研究表达 (2.1) 的第三方 preT 细胞。 CD19-CAR，(2.2) CD19-CAR 和细胞因子，或 (2.3) CD19-CAR/WT1 特异性 TCR。 已经利用供体来源的 CD19-CAR T 细胞或第三方开展了两项同种异体 HCT 临床试验 preT 细胞，加上我们在 CAR T 细胞方面丰富的临床经验，将有助于 转化拟议的临床前研究以减少异基因 HCT 患者的复发 使用第三方 CAR T 细胞治疗血液恶性肿瘤。