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

• 批准号: 10211972
• 负责人: Chintan Parekh
• 金额: $ 43.59万
• 依托单位: CHILDREN'S HOSPITAL OF LOS ANGELES
• 依托单位国家: 美国
• 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/10211972
• 关键词: 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; 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; Lead; 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; RNA; Recovery; Recurrence; Regulation; Relapse; Repression; Role; Safety; Secondary to; Signal Transduction; Solid; Solid Neoplasm; 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; Tumor stage; Up-Regulation; Work; alpha-Thalassemia; anti-cancer; base; cancer cell; chimeric antigen receptor; chimeric antigen receptor T cells; curative treatments; cytokine; cytotoxic; engineered stem cells; exhaustion; fighting; gain of function; graft vs host disease; human data; humanized mouse; improved; improved outcome; in vivo; innovation; insight; knock-down; leukemia; leukemia relapse; loss of function; mouse model; neutralizing antibody; novel; novel strategies; overexpression; peripheral blood; preclinical efficacy; progenitor; programs; recruit; recurrent infection; response; single-cell RNA sequencing; transcription factor; tumor microenvironment

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细胞系转录因子（TF）BCL 11 B的过表达作为一种新的策略 增强：1）异基因造血干细胞移植（HSCT）后的T细胞重建，和2） 抗癌嵌合抗原受体（CAR）T细胞的功效。造血干细胞移植是治疗多种白血病的有效方法 单独或作为CAR后巩固治疗。然而，从供体造血细胞中产生的T细胞 干细胞和祖细胞（HSPC）需要数月的时间，使危及生命的感染和白血病复发 HSCT的主要挑战。虽然CAR T细胞在CD 19+白血病中诱导高缓解率，但T细胞免疫功能低下。 功能和持久性以及由于肿瘤微环境的抑制而导致的T细胞耗竭仍然是主要的 CAR T细胞在白血病和实体瘤中的治疗效果的障碍。物种相关差异 TF对T细胞分化的调节以及对人类T细胞分化机制的认识不足， T细胞分化是开发增强T细胞分化和 功能肿瘤抑制因子TF Bcl 11b是抑制替代（非T）谱系潜能所必需的 但在T细胞分化的初始阶段，在诱导T细胞系基因表达中不起作用。 小鼠HSPC的分化。相比之下，我们发现BCL 11 B对T细胞的诱导和T细胞的增殖都是至关重要的。 在人类T细胞分化的初始阶段， 分化我们现在有了新的初步体外数据，慢病毒BCL 11B过表达：1）加速 来自人HSPC的T细胞分化，包括成熟T细胞的产生，和2）增强功能， 促进分化成具有中枢记忆表型的细胞，并延迟人T细胞的耗竭。 功能性、Chip-Seq和单细胞RNA-Seq数据的综合分析揭示了NOTCH 3和IRF 8是 人类中BCL 11B的物种特异性候选靶点。值得注意的是，BCL 11B过表达研究没有 由于毒性，在鼠HSPC中是可能的。基于这些数据，我们假设移植HSPC 工程化以过表达BCL 11B将增强HSCT后T细胞重建。BCL 11B过表达将 通过增强CAR T细胞的功能和持久性以及改善疲劳来提高CAR T细胞的功效。 我们将通过以下目的来检验这一假设：1.1）确定BCL 11B对T细胞的表观遗传效应， 细胞基因和BCL 11 B介导的人T细胞中NOTCH 3（1.2）和IRF 8（1.3）的调节作用 分化1.4)确定过表达人HSPC的BCL 11B用于增强人HSPC后增殖的功效。 人源化小鼠模型中的HSCT T细胞重建，和2）确定BCL 11B过表达的作用 对白血病和神经母细胞瘤中人CAR T细胞的抗癌功效、持久性和耗竭的影响 模型这些研究可能揭示BCL 11B的新功能，并导致BCL 11B工程细胞疗法 改善白血病和实体瘤的治疗效果。这项建议是创新的，因为它建立在我们的工作基础上。 定义BCL 11B在人类中的物种特异性作用，以解决HSCT和CAR T细胞治疗中的关键障碍。