MODULATION OF CD5 SIGNALING TO ENHANCE ADOPTIVE T-CELL THERAPIES FOR CANCER

调节 CD5 信号传导以增强癌症过继性 T 细胞疗法

基本信息

批准号：
10609068
负责人：
Marco Ruella
金额：
$ 51.7万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-12 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10609068
关键词：
Affinity Antigens B lymphoid malignancy B-Cell Leukemia B-Lymphocytes Behavior CBL gene CD19 gene CD8-Positive T-Lymphocytes CRISPR/Cas technology CTAG1 gene Calcium Signaling Cell Death Induction Cell physiology Cells Chronic Clinical Clinical Trials Complex Data Development Disease remission Disinhibition ERBB2 gene Exposure to Flow Cytometry Goals Grant Hematology Human Immune checkpoint inhibitor Immunocompetent Immunotherapy In Vitro Investigation Knock-out Knowledge Lymphoma Malignant Neoplasms Malignant neoplasm of ovary Malignant neoplasm of pancreas Malignant neoplasm of prostate Mediating Mediator Mission Modeling Multiple Myeloma Mus Outcome PD-1/PD-L1 PTPN6 gene Pathway interactions Patients Peripheral Persons Phosphorylation Pre-Clinical Model Process Production Proliferating Public Health Receptor Signaling Refractory Relapse Reporting Research Role Safety Scientific Advances and Accomplishments Signal Pathway Signal Transduction Solid Solid Neoplasm T cell therapy T-Cell Activation T-Cell Leukemia T-Cell Lymphoma T-Cell Receptor T-Lymphocyte Technology Testing Work Xenograft Model Xenograft procedure antitumor effect cancer immunotherapy cancer therapy chimeric antigen receptor T cells clinical practice clinically relevant cytokine cytotoxicity digital effector T cell engineered T cells exhaustion experience first-in-human growth factor receptor-bound protein 2 human model immunological synapse immunoregulation improved in vivo innovation leukemia/lymphoma melanoma mesothelin mouse model next generation novel patient subsets performance tests programmed cell death protein 1 refractory cancer release of sequestered calcium ion into cytoplasm response safety testing scavenger receptor side effect single-cell RNA sequencing transcription factor tumor tumor microenvironment tumor-immune system interactions

项目摘要

PROJECT SUMMARY Adoptive T-cell immunotherapies and, in particular, chimeric antigen receptor T cells (CART) generated unprecedented responses in patients with highly refractory CD19+ B cell malignancies. However, only a limited number of patients treated with anti-CD19 CART will experience prolonged remission while most patients either do not respond or eventually relapse. Moreover, in the setting of solid cancers, CART cells have generally been disappointing. Increasing CART effector function against cancer would represent a vertical improvement in the field of adoptive T-cell immunotherapy. In this proposal, we aim at increasing the anti-tumor efficacy of CAR T cells by reducing the inhibition of CAR activation. Current approaches to enhance CART efficacy focus on reducing long-term T cell exhaustion by targeting the PD-1/PD-L1 axis. However, there is a lack of investigation on strategies to enhance early CAR and T cell receptor (TCR) signaling, which is critical for effective tumor killing, especially in the setting of the immunosuppressive tumor microenvironment. To this goal, we studied the scavenger receptor CD5. CD5 associates with the TCR complex and inhibits its activation through several mediators, including SHP-1, CBL, CBL-B, and GRB2. The central hypothesis of this grant is that CD5 deletion increases the anti-tumor efficacy of engineered T cells through enhancement of TCR signaling. In our preliminary work, we have found that CRISPR-Cas9 CD5 knock out in CART cells enhances their anti-tumor activity in xenograft models of B-cell leukemia (CART19) and T-cell lymphoma (CART5). Moreover, we demonstrated that, upon stimulation, CD5 knocked-out CART cells show higher phosphorylation of key CAR signaling mediators as compared to wild-type. In Aim#1, we will test the hypothesis that the deletion of CD5 in T cells broadly enhances the anti-tumor efficacy of adoptive T-cell immunotherapies, by studying CD5 knockout in the setting of CAR T cells for solid tumors. In Aim#2, we will test the hypothesis that CD5 deletion augments T-cell anti-tumor activity by disinhibiting CAR signaling by performing in vitro and in vivo mechanistic studies. We will also test the safety of this approach. This hypothesis-driven proposal is highly innovative as it proposes the study of a novel immunomodulatory target - CD5 - and uses cutting edge technologies like digital spatial profiling and single-cell RNA sequencing to investigate CD5 deleted CAR T-cell in vivo in both human and murine models. A successful outcome of the proposed investigations is expected to impact the field of cancer immunotherapy significantly, providing a rational and generalizable approach to improving T-cell immunotherapies and ultimately drive the development of a first-in-human clinical trial of CD5 knocked-out CART for refractory cancers.

项目概要过继性 T 细胞免疫疗法，特别是嵌合抗原受体 T 细胞 (CART) 在高度难治性 CD19+ B 细胞恶性肿瘤患者中出现了前所未有的反应。然而，只有有限的接受抗 CD19 CART 治疗的患者数量将经历长期缓解，而大多数患者要么没有反应或最终复发。此外，在实体癌的治疗中，CART 细胞通常被令人失望。增加针对癌症的 CART 效应器功能将代表癌症治疗的垂直改进过继性T细胞免疫治疗领域。在这个提案中，我们的目标是提高 CAR T 的抗肿瘤功效通过减少 CAR 激活的抑制来抑制细胞。目前增强 CART 功效的方法主要集中在通过靶向 PD-1/PD-L1 轴来减少长期 T 细胞耗竭。但缺乏调查增强早期 CAR 和 T 细胞受体 (TCR) 信号传导的策略，这对于有效杀死肿瘤至关重要，尤其是在免疫抑制肿瘤微环境的情况下。为了这个目标，我们研究了清道夫受体CD5。 CD5 与 TCR 复合物结合并通过多种方式抑制其激活介质，包括 SHP-1、CBL、CBL-B 和 GRB2。本次资助的中心假设是 CD5 缺失通过增强 TCR 信号传导来提高工程化 T 细胞的抗肿瘤功效。在我们的初步工作中，我们发现 CART 细胞中的 CRISPR-Cas9 CD5 敲除增强了它们的抗肿瘤活性 B 细胞白血病 (CART19) 和 T 细胞淋巴瘤 (CART5) 的异种移植模型。此外，我们证明了，刺激后，CD5 敲除的 CART 细胞表现出关键 CAR 信号传导介质的更高磷酸化，如下所示与野生型相比。在目标 #1 中，我们将检验 T 细胞中 CD5 缺失广泛增强的假设通过研究 CAR T 环境下的 CD5 敲除来了解过继性 T 细胞免疫疗法的抗肿瘤功效实体瘤细胞。在目标#2 中，我们将检验 CD5 缺失增强 T 细胞抗肿瘤活性的假设通过进行体外和体内机制研究来抑制 CAR 信号传导。我们也会测试安全性这种方法。这个假设驱动的提案具有高度创新性，因为它提出了一项新颖的研究免疫调节靶标 - CD5 - 并使用数字空间分析和单细胞等尖端技术 RNA 测序可在人类和小鼠模型中研究体内 CD5 缺失的 CAR T 细胞。一个成功的拟议研究的结果预计将对癌症免疫治疗领域产生重大影响，提供合理且可推广的方法来改善 T 细胞免疫疗法并最终推动开发针对难治性癌症的 CD5 敲除 CART 的首次人体临床试验。