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-Linege转录因子（TF）BCL11B的过表达作为一种新型策略增强：1）同种异性造血干细胞移植（HSCT）和2）抗癌嵌合抗原受体（CAR）T细胞的功效。 HSCT是许多白血病的治疗疗法本身或作为车后整合疗法。但是，供体造血的T细胞产生茎和祖细胞（HSPC）需要数月才能使生命威胁性感染和白血病复发 HSCT面临的主要挑战。而汽车T细胞在CD19+白血病中诱导高缓解率，而T细胞不良功能，持久性以及由于肿瘤微环境抑制而导致的T细胞疲惫汽车T细胞在白血病和实体瘤中的治愈功效的障碍。物种相关的差异通过TF调节T细胞分化以及对人T细胞机制的不良理解差异化已成为发展T细胞差异化和的方法的障碍功能。抑制替代（非T）谱系电位需要肿瘤抑制剂TF BCl11b 但是在T细胞的初始阶段的T-Lineage基因表达中不起作用鼠HSPC的分化。相比之下，我们表明Bcl11b对于T-的诱导至关重要人类T细胞初始阶段期间的血统计划和抑制替代血统计划分化。现在，我们有新型的初步体外数据，可以慢性病Bcl11b过表达：1） T细胞与人类HSPC的分化，包括成熟T细胞的产生，2）增强功能，通过中央记忆表型促进分化为细胞，并延迟了人类T细胞的精疲力尽。功能，芯片序列和单细胞RNA-seq数据的综合分析揭示了Notch3和IRF8为人类BCl11b的物种特定候选靶标。值得注意的是，Bcl11b的过表达研究尚未由于毒性，在鼠HSPC中可能发生了可能。基于这些数据，我们假设HSPC的移植设计为过表达BCL11b的设计将增强HSCT后T细胞重建。 Bcl11b过表达将通过增强其功能，持久性和缓解疲惫，提高了汽车T细胞的功效。我们将通过以下目的检验假设：1.1）确定Bcl11b对T-的表观遗传作用细胞基因以及Bcl11b介导的Notch3（1.2）和IRF8（1.3）在人类T细胞中的作用分化。 1.4）定义Bcl11b过表达人类HSPC的功效 HSCT T细胞重建在人源性小鼠模型中，2）定义BCL11b过表达的效果关于抗癌的功效，持久性和人类汽车T细胞在白血病和神经母细胞瘤中的衰竭型号。这些研究可以揭示BCL11B的新功能并导致BCL11B工程细胞疗法这可以改善白血病和实体瘤的预后。该建议具有创新性，因为它建立在我们的工作基础上定义BCL11b在人类中的特定物种影响，以解决HSCT和CAR T细胞疗法中的关键障碍。