Bcl11b: A Master Transcription Factor Controlling Human NK Cell Development

Bcl11b：控制人类 NK 细胞发育的主转录因子

• 批准号: 10295051
• 负责人: Frank Cichocki
• 金额: $ 31万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10295051
• 关键词: ATAC-seq; Adoptive Cell Transfers; Adoptive Transfer; Advanced Malignant Neoplasm; Automobile Driving; CD3 Antigens; CD8-Positive T-Lymphocytes; Cell Differentiation process; Cell Line; Cell Maturation; Cell physiology; Cell-Mediated Cytolysis; Cells; ChIP-seq; Characteristics; Complement; Cytokine Receptors; Cytokine Signaling; Cytomegalovirus; Data; Development; Drops; Effector Cell; Enhancers; Epigenetic Process; Exhibits; Flow Cytometry; Foundations; Funding; Gene Expression Profile; Genes; Genetic Transcription; Genomic Segment; Hematopoietic stem cells; Human; Immune; Immune response; Immunologic Memory; Immunotherapeutic agent; Immunotherapy; In Vitro; Individual; Inflammatory; Killer Cells; Knowledge; Literature; Lymphocyte; Mediating; Modeling; Mus; Natural Killer Cells; Ontology; Patients; Phenotype; Process; Property; Receptor Signaling; Role; Signaling Protein; T-Cell Development; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Tumor Immunity; Umbilical Cord Blood; Viral; Work; adaptive immune response; antiviral immunity; base; cancer cell; chemokine; cytokine; cytotoxic; design; experimental study; improved; in vivo; in vivo Model; induced pluripotent stem cell; interest; knock-down; overexpression; peripheral blood; pre-clinical; programs; promoter; receptor; seropositive; transcription factor; transcriptome sequencing; tumor

PROJECT SUMMARY/ABSTRACT Natural killer (NK) cells are innate lymphocytes that mediate cellular cytotoxicity against virally infected and malignant cells. Upon activation, they also release chemokines and cytokines that help prime and coordinate the adaptive immune response. Because of these functional properties, there is continued interest in developing NK cell products for immunotherapy. However, this effort is hindered by our limited understanding of human NK cell differentiation and the transcription factor networks that regulate this process. In work supported by current K99/R00 funding, we generated a considerable amount of preliminary data comprehensively characterizing the transcriptional and epigenetic landscapes in sorted NK and CD8+ T cell subsets isolated from cytomegalovirus (CMV) seropositive donors. To complement these analyses, we performed flow cytometry-based analyses of transcription factor expression and ChIP-seq on sorted peripheral blood NK cell subsets to construct transcription factor networks that potentially regulate key steps in NK cell differentiation. The culmination of this work revealed a central role for Bcl11b. This was completely unexpected given several detailed studies in mice showing that Bcl11b expression is restricted to the T, NKT, and ILC2 lineages and acts to enforce T cell identity. Our preliminary results also revealed a reciprocal relationship between Runx2 and Bcl11b, with Runx2 regulating a set of transcription factors enriched in immature NK cells and Bcl11b regulating genes enriched throughout canonical NK cell maturation and in adaptive NK cells exhibiting a T cell-like gene expression signature. In this application we will test the hypothesis that Runx2 is a major hub that enforces the epigenetic identity of immature CD56bright NK cells, while Bcl11b suppresses the Runx2-directed transcriptional program to drive canonical NK cell differentiation, maturation, and cytotoxic effector function. We also posit that high levels of Bcl11b contribute to the T cell-like features of CMV-induced adaptive NK cells. We propose two independent aims to test our hypotheses. In the first aim, we will use overexpression and knockdown strategies in sorted NK cell subsets to define the roles of Bcl11b and Runx2 during canonical NK cell differentiation. These results will guide subsequent experiments designed to determine whether Bcl11b overexpression in hematopoietic progenitor cells (HPCs) promotes NK cell maturation and effector function both in vitro and after adoptive transfer into mice with established tumors. In the second aim, we will determine the role of Bcl11b in promoting T cell-associated features of CMV-induced adaptive NK cells and test the hypothesis that high levels of Bcl11b along with activating receptor and cytokine receptor stimulation drives adaptive NK cell differentiation and maturation. We will also test the hypothesis that adaptive NK cells mediate superior antitumor function in vivo relative to canonical NK cells. Results generated from this proposal will define the role of Bcl11b in promoting canonical and adaptive NK cell differentiation, determine whether Bcl11b overexpression enhances NK cell-mediated antitumor activity, and definitively test the antitumor function of adaptive NK cells.

项目摘要/摘要 天然杀伤（NK）细胞是先天淋巴细胞，可针对病毒感染和 恶性细胞。激活后，它们还释放趋化因子和细胞因子，有助于启用和协调 自适应免疫反应。由于具有这些功能性能，因此继续兴趣开发NK 用于免疫疗法的细胞产品。但是，我们对人类NK细胞的理解有限地阻碍了这一努力 分化和调节此过程的转录因子网络。在当前支持的工作中 K99/R00资金，我们生成了大量的初步数据 从巨细胞病毒分离的分类NK和CD8+ T细胞子集中的转录和表观遗传景观 （CMV）血清阳性供体。为了补充这些分析，我们进行了基于流式细胞仪的分析 在排序的外周血NK细胞子集上的转录因子表达和芯片序列构建转录 因子网络可能调节NK细胞分化的关键步骤。这项工作的高潮揭示了 BCL11b的核心角色。鉴于在小鼠中进行了一些详细研究，这是完全出乎意料的。 Bcl11b的表达仅限于t，nkt和ILC2谱系，并作用于强制T细胞身份。我们的 初步结果还揭示了Runx2和Bcl11b之间的相互关系，Runx2调节A 富含NK细胞和BCL11B调节基因的转录因子集遍布遍及整个基因 典型的NK细胞成熟和在自适应NK细胞中表现出T细胞样基因表达特征。在这个 应用我们将测试以下假设：Runx2是一个主要的枢纽，可实施未成熟的表观遗传身份 CD56Bright NK细胞，而BCL11B抑制了Runx2定向的转录程序以驱动规范NK 细胞分化，成熟和细胞毒性效应子功能。我们还认为，高水平的Bcl11b贡献 CMV诱导的自适应NK细胞的T细胞样特征。 我们提出了两个独立的目标来检验我们的假设。在第一个目标中，我们将使用过表达 以及分类的NK单元子集中的敲低策略，以定义BCL11B和RUNX2在规范期间的作用 NK细胞分化。这些结果将指导后续实验旨在确定BCL11b是否 造血祖细胞（HPC）的过表达促进NK细胞的成熟和效应子功能 体外和过继转移到具有既定肿瘤的小鼠中。在第二个目标中，我们将确定 BCl11b在促进CMV诱导的自适应NK细胞的T细胞相关特征中的作用并检验假设 高水平的Bcl11b以及激活受体和细胞因子受体刺激驱动自适应NK细胞 分化和成熟。我们还将测试自适应NK细胞介导上抗肿瘤的假设 相对于规范NK细胞的体内功能。该提案产生的结果将定义Bcl11b的作用 在促进规范和适应性NK细胞分化时，确定BCL11B是否会增强 NK细胞介导的抗肿瘤活性，并确定测试自适应NK细胞的抗肿瘤功能。