BCL11B activation as an approach for enhancing the efficacy of immunotherapy

BCL11B 激活作为增强免疫疗法疗效的方法

基本信息

批准号：
10582696
负责人：
Chintan Parekh
金额：
$ 49.92万
依托单位：
CHILDREN'S HOSPITAL OF LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-11 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10582696
关键词：
Acceleration Address Allogenic B-Cell Acute Lymphoblastic Leukemia Binding Bone Marrow Transplantation CAR T cell therapy CD19 gene Cell Differentiation process Cell Therapy Cell physiology Cells ChIP-seq Chromatin Complex Consolidation Therapy Data Development Disease remission Engineering Epigenetic Process Frequencies Gene Expression Generations Genes Genetic Transcription Goals Hematopoiesis Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human Immune Immune system Immunity Immunotherapy Impairment In Vitro Infection Kinetics Knowledge Lentivirus Leukocytes Life Malignant Neoplasms Mature T-Lymphocyte Mediating Memory Methods Modeling Monitor Mus NOTCH1 gene NOTCH3 gene Neuroblastoma Nucleosomes Outcome Patients Phenotype Process Production Proliferating Recovery Recurrence Regulation Relapse Repression Role Safety Secondary to Signal Transduction Solid Solid Neoplasm Specific qualifier value T cell differentiation T cell reconstitution T cell regulation T-Cell Activation T-Cell Development T-Lymphocyte Testing Thymus Gland Toxic effect Transplantation Tumor Suppressor Proteins Up-Regulation Work alpha-Thalassemia anti-cancer cancer cell cell motility chimeric antigen receptor chimeric antigen receptor T cells curative treatments cytokine cytotoxic efficacy evaluation engineered stem cells exhaustion fighting gain of function graft vs host disease hematopoietic transplantation human data humanized mouse improved improved outcome in vivo innovation insight knock-down leukemia leukemia relapse leukemia treatment loss of function mouse model neutralizing antibody novel novel strategies overexpression peripheral blood preclinical efficacy progenitor programs recruit response single-cell RNA sequencing transcription factor tumor microenvironment

项目摘要

Our goal is to investigate overexpression of the T-lineage transcription factor (TF) BCL11B as a novel strategy to enhance: 1) T-cell reconstitution after allogeneic hematopoietic stem cell transplantation (HSCT), and 2) the efficacy of anticancer chimeric antigen receptor (CAR) T-cells. HSCT is a curative therapy for many leukemias by itself or as a post-CAR consolidation therapy. However, the generation of T-cells from donor hematopoietic stem and progenitor cells (HSPC) takes many months making life threatening infections and leukemia relapse major challenges in HSCT. While CAR T-cells induce high remission rates in CD19+ leukemias, poor T-cell function and persistence and T-cell exhaustion due to inhibition by the tumor microenvironment remain major obstacles to the curative efficacy of CAR T-cells in leukemia and solid tumors. Species related differences in the regulation of T-cell differentiation by TF and the poor understanding of mechanisms in human T-cell differentiation have been hurdles to the development of approaches to enhance T-cell differentiation and function. The tumor suppressor TF Bcl11b is required for the repression of alternative (non-T) lineage potentials but does not play a role in the induction of T-lineage gene expression during the initial stages of T-cell differentiation of murine HSPC. In contrast, we showed that BCL11B is critical for both the induction of the T- lineage program and repression of alternative lineage programs during the initial stages of human T-cell differentiation. We now have novel preliminary in vitro data that lentiviral BCL11B overexpression: 1) expedites T-cell differentiation from human HSPC including the generation of mature T-cells, and 2) enhances the function, promotes differentiation into cells with a central memory phenotype, and delays exhaustion of human T-cells. Integrated analysis of functional, Chip-Seq, and single cell RNA-Seq data revealed NOTCH3 and IRF8 as species specific candidate targets of BCL11B in humans. Of note, BCL11B overexpression studies have not been possible in murine HSPC due to toxicity. Based on these data, we hypothesize that transplantation of HSPC engineered to overexpress BCL11B will enhance post-HSCT T-cell reconstitution. BCL11B overexpression will increase the efficacy of CAR T-cells by enhancing their function and persistence and ameliorating exhaustion. We will test the hypothesis through the following aims: 1.1) Determine the epigenetic effects of BCL11B on T- cell genes and the role of BCL11B mediated regulation of NOTCH3 (1.2) and IRF8 (1.3) in human T-cell differentiation. 1.4) Define the efficacy of BCL11B overexpressing human HSPC for the enhancement of post- HSCT T-cell reconstitution in humanized mouse models, and 2) Define the effects of BCL11B overexpression on anti-cancer efficacy, persistence, and exhaustion of human CAR T-cells in leukemia and neuroblastoma models. These studies could reveal new functions of BCL11B and lead to BCL11B engineered cell therapies that improve outcomes in leukemia and solid tumors. This proposal is innovative because it builds on our work defining species specific effects of BCL11B in humans to address key barriers in HSCT and CAR T-cell therapy.

我们的目标是研究 T 谱系转录因子 (TF) BCL11B 的过度表达作为一种新策略增强：1) 异基因造血干细胞移植 (HSCT) 后 T 细胞重建，2) 嵌合抗原受体（CAR）T 细胞的抗癌功效。 HSCT 是许多白血病的治愈方法单独或作为 CAR 后巩固疗法。然而，从供体造血细胞生成 T 细胞干细胞和祖细胞 (HSPC) 需要数月时间才能导致危及生命的感染和白血病复发 HSCT 的主要挑战。虽然 CAR T 细胞可诱导 CD19+ 白血病的高缓解率，但较差的 T 细胞肿瘤微环境抑制导致的功能和持久性以及 T 细胞耗竭仍然是主要因素 CAR T细胞在白血病和实体瘤中的疗效存在障碍。与物种相关的差异 TF 调节 T 细胞分化以及对人类 T 细胞机制的了解不足分化一直是增强 T 细胞分化方法开发的障碍，功能。肿瘤抑制因子 TF Bcl11b 是抑制替代（非 T）谱系电位所必需的但在 T 细胞初始阶段不参与 T 谱系基因表达的诱导小鼠HSPC的分化。相比之下，我们发现 BCL11B 对于 T- 的诱导至关重要。人类 T 细胞初始阶段的谱系程序和替代谱系程序的抑制差异化。我们现在有新的初步体外数据表明慢病毒 BCL11B 过度表达：1) 加速来自人类 HSPC 的 T 细胞分化，包括生成成熟 T 细胞，2) 增强功能，促进分化为具有中央记忆表型的细胞，并延缓人类 T 细胞的耗竭。对功能、Chip-Seq 和单细胞 RNA-Seq 数据的综合分析显示 NOTCH3 和 IRF8 人类中 BCL11B 的物种特异性候选靶标。值得注意的是，BCL11B 过度表达研究尚未由于毒性，在小鼠 HSPC 中可能存在这种情况。根据这些数据，我们假设 HSPC 移植过表达 BCL11B 将增强 HSCT 后 T 细胞的重建。 BCL11B 过度表达会通过增强 CAR T 细胞的功能和持久性以及改善疲劳来提高其功效。我们将通过以下目标检验该假设： 1.1) 确定 BCL11B 对 T- 的表观遗传效应细胞基因以及 BCL11B 介导的人 T 细胞中 NOTCH3 (1.2) 和 IRF8 (1.3) 调节的作用差异化。 1.4) 定义 BCL11B 过表达人 HSPC 对增强后人源化小鼠模型中的 HSCT T 细胞重建，以及 2) 定义 BCL11B 过表达的影响人类 CAR T 细胞在白血病和神经母细胞瘤中的抗癌功效、持久性和耗竭的研究模型。这些研究可能揭示 BCL11B 的新功能并导致 BCL11B 工程细胞疗法改善白血病和实体瘤的治疗结果。该提案具有创新性，因为它建立在我们的工作基础上确定 BCL11B 对人类的物种特异性影响，以解决 HSCT 和 CAR T 细胞治疗中的关键障碍。