IL-2 Family Cytokines and Receptors-- Mechanisms of Regulation & Action

IL-2 家族细胞因子和受体——调节机制

• 批准号: 10929101
• 负责人: Warren J Leonard
• 金额: $ 184.75万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10929101
• 关键词: 3-Dimensional; Adoptive Transfer; Adult T-Cell Leukemia/Lymphoma; Agonist; Allergic; Attenuated; Autoimmune Diseases; B-Lymphocytes; BCL6 gene; Binding; Biological; Biological Process; CD27 Antigens; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell Differentiation process; Cell physiology; Cells; Closure by clamp; Collaborations; Coupled; Cytokine Receptors; DNA sequencing; Data; Defect; Dendritic Cells; Development; Disease; EGR2 gene; Elements; Enhancers; Event; Eyelid structure; Family; Functional disorder; Gene Expression; Genes; Genetic; Genetic Enhancer Element; Genetic Transcription; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Helminths; Helper-Inducer T-Lymphocyte; Homing; Host Defense; Human; Human T-lymphotropic virus 1; IL2 gene; IL2RA gene; IL7 gene; IRF4 gene; Immune; Immune response; Immunologic Deficiency Syndromes; Immunologics; Inbred BALB C Mice; Induction of Apoptosis; Inflammatory; Interleukin 2 Receptor; Interleukin 2 Receptor Gamma; Interleukin 4 Receptor; Interleukin-13; Interleukin-15; Interleukin-2; Interleukin-4; Interleukin-9; Knock-in Mouse; Malignant - descriptor; Malignant Neoplasms; Maps; Modeling; Molecular; Mus; Mutate; Names; Natural Killer Cells; PRDM1 gene; Pathogenicity; Pathologic; Phenotype; Physiological; Population; Proliferating; Protein Family; Proteins; Proto-Oncogene Proteins c-jun; Publishing; Receptor Signaling; Regulation; Regulatory T-Lymphocyte; Reporting; Repression; Response Elements; Role; STAT protein; STAT1 gene; STAT3 gene; STAT5B gene; Signal Induction; Signal Transduction; Stat5 protein; System; T-Cell Activation; T-Lymphocyte; Therapeutic; Time; Transcription Factor AP-1; Transgenic Mice; Variant; Virus; X-Linked Severe Combined Immunodeficiency; adult T-cell leukemia cell; biomarker discovery; cell type; chimeric antigen receptor T cells; chromatin immunoprecipitation; cytokine; dextran sulfate sodium induced colitis; dimer; epigenome; exhaustion; genome-wide; graft vs host disease; human disease; immune system function; in vivo; insight; interleukin-15 receptor; interleukin-21; interleukin-21 receptor; melanoma; mimetics; monocyte; multiple omics; mutant; novel; novel diagnostics; novel strategies; novel therapeutics; pathogen; promoter; protein activation; protein expression; receptor; receptor expression; response; three dimensional structure; transcription factor; transcriptome sequencing; tumor; user-friendly

The IL-2 receptor and related cytokine/cytokine receptor systems are being studied to understand the T cell immune response in normal and disease. After T-cell activation, the magnitude and duration of the response is controlled by the amount of IL-2 produced, levels of IL-2 receptors, and the time course of their induction. IL-2Ra expression is highly expressed by cells infected with HTLV-I, the cause of adult T cell leukemia (ATL). There are 3 chains of the receptor: IL-2Ra, IL-2Rb, and gc, with IL-2Ra and IL-2Rb regulated at the level of transcription. gc is shared by the IL-4, IL-7, IL-9, IL-15, and IL-21 receptors and is mutated in XSCID. We study the signals induced by these cytokines, particularly STAT proteins and the mechanisms by which they regulate target genes. Given our prior data that Stat5a or Stat5b transgenic mice develop tumors, consistent with STAT5 being implicated in malignant transformation and elevated in a range of human tumors, this has relevance for both normal and pathological states. Moreover, humans and mice with altered STAT protein expression or activation have a range of immunological defects. T helper cell differentiation is critical for normal immune responses, with Th1 differentiation important for host defense to viruses and other intracellular pathogens, Th2 differentiation vital in allergic disorders/helminths, and Th17 differentiation vital in inflammatory disorders. We previously showed that IL-2 is important for Th2 differentiation and that IL-2 induces IL-4 receptor expression in a STAT5-dependent manner and controls priming for Th2 differentiation. Moreover, using genome-wide chromatin immunoprecipitation coupled to DNA sequencing (ChIP-Seq), we previously found regulation of Th2 differentiation via STAT5A and STAT5B and extended these findings to show that IL-2 via STAT5 induces IL-12Rb2, which is critical for Th1 differentiation and that IL-2 via STAT5 also regulates T-bet. we also had elucidated mechanism by which IL-2 inhibits Th17 differentiation. We also reported a critical role of IL-2 in Th9 differentiation, with IL2 inducing STAT5 binding to the Il9 promoter and IL-2 and IL-21 having opposing actions in Th9 differentiation based on induction of BCL6 by IL-21 but repression by IL-2. With Dr. K. Christopher Garcia (Stanford), we previously studied novel IL-2 variant partial agonists that function as "receptor signaling clamps. We previously showed that one variant, H9-RETR, prolonged survival in a model of graft-versus-host disease and blocked spontaneous proliferation of smoldering ATL cells. This receptor-clamping approach might be a general mechanism-based strategy. IL-2 REH was reported as a partial agonist that specifically expanded Treg rather than CD8 T cells, with benefit in DSS colitis, and we studied another partial agonist that promoted a TCSM phenotype of CD8 T cells, with enhanced efficacy in adoptive transfer treatment of B16 melanoma and in a second generated CAR T antitumor model. In the latter, we had discovered by STAT5 is a promoter of T cell exhaustion, with attenuated STAT5 activation resulting in a TSCM phenotype and elucidated underlying mechanisms. In the past year, we have pursued studies of other IL-2 partial agonists and IL-21 mimics. IL-21 has broad actions on T- and B-cells, and we previously reported that it also induces apoptosis of conventional dendritic cells. ChIP-Seq analysis had revealed genome-wide binding competition between GM-CSF-induced STAT5 and IL-21-induced STAT3, and we previously elucidated roles for STAT1 vs. STAT3 in IL-21 signaling inT cells. Previously, we also showed that IL-21 regulates expression of the Prdm1 gene that encodes BLIMP1 via a response element that depends on STAT3 and IRF4 and subsequently discovered that in contrast to its known ability to cooperate with PU.1 in B cells to act via Ets-IRF composite elements (EICEs), IRF4 cooperates with BATF/JUN family proteins to act via novel AP1-IRF composite elements (AICEs) in T cells, as well as in B cells. In other studies, we previously studied the biological roles of Egr1 and Egr2 and elucidated some non-immunological roles for Egr1, demonstrating that this transcription factor has a genetic-background dependent effect on eyelid development-- being required for such development on the BALB/c background but not on the C57BL/6 background. In collaborative studies, we have advanced studies of Egr family proteins and this year a collaborative study was published on the role of EGR family proteins in the homing and pathogenicity of Th17 cells in the CNS and we have also collaborated with Jim Salzer related to roles of EGR proteins in the CNS. Previously, we studied the biological significance of STAT5 tetramerization in vivo by generating mice expressing mutant forms of STAT5A and STAT5B that could form dimers but not tetramers and previously reported modeling of the 3-dimensional structure of the tetramer, and we also previously reported the basis for defective NK cell development in the double knockin mice, demonstrating a critical role for STAT5 tetramers for the survival of NK cells. In the current year, we now have extended our studies of STAT5 tetramers and reported their role related to the regulation of monocyte differentiation. We also previously globally characterized super-enhancers regulated by IL-2-activated STAT5 and IL-21-activated STAT3 and their relationship to highly inducible genes and had found that the Il2ra gene contains the most highly ranked STAT5-dependent super enhancer. Using CRISPR-Cas9 technology, we previously functionally dissected the elements of this super-enhancer, providing key new insights into the molecular regulation of the Il2ra in particular and super-enhancers more generally, and in the current year, we have significantly extended these studies, clarifying the differential importance of different super-enhancer elements in different cell types. Moreover, this year we contributed to studies of IL-4 mimetics in the distinctive activation of type 1 IL-4 receptors, with RNA-seq indicating similar effects of neo-4 to those of IL-4 and IL-13 on monocytes. We also have reported in a collaborative study an efficient new approach, ChIATAC, for the multiomics mapping of 3D epigenomes and reported a new user-friendly R Shiny application for RNA-sequencing analysis and biomarker discovery. Overall, these studies enhance our understanding of mechanisms by which gc family cytokines regulate gene expression and biological processes and are relevant to normal and pathological immune cell function, including in disease states, with potential therapeutic implications.

正在研究IL-2受体和相关的细胞因子/细胞因子受体系统，以了解正常和疾病中的T细胞免疫反应。 T细胞激活后，响应的幅度和持续时间由产生的IL-2的量，IL-2受体的水平及其诱导的时间过程控制。 IL-2RA表达由感染HTLV-I的细胞高度表达，这是成年T细胞白血病（ATL）的原因。有3个受体的链：IL-2RA，IL-2RB和GC，在转录水平下调节IL-2RA和IL-2RB。 GC由IL-4，IL-7，IL-9，IL-15和IL-21受体共享，并在XSCID中突变。我们研究了这些细胞因子，尤其是Stat蛋白及其调节靶基因的机制的信号。鉴于我们先前的数据是STAT5A或STAT5B转基因小鼠会发展为肿瘤，这与STAT5与恶性转化有关，并且在一系列人类肿瘤中升高，这与正常状态和病理状态均相关。此外，具有改变统计蛋白表达或激活的人类和小鼠具有一系列免疫缺陷。 T辅助细胞分化对于正常的免疫反应至关重要，Th1分化对于宿主防御对病毒和其他细胞内病原体，TH2分化在过敏性疾病/蠕虫中至关重要，而Th17在炎症性疾病中至关重要。我们先前表明IL-2对于Th2分化很重要，并且IL-2以STAT5依赖性方式诱导IL-4受体表达，并控制Th2分化的启动。此外，使用与DNA测序结合（CHIP-SEQ）耦合的全基因组染色质免疫沉淀，我们先前发现了通过Stat5a和Stat5B对Th2分化的调节，并扩展了这些发现以表明IL-2通过STAT5通过STAT5诱导IL-12RB2，这对TH1分化以及IL-2通过Stat5 Chart5也很重要。我们还阐明了IL-2抑制Th17分化的机制。我们还报道了IL-2在Th9分化中的关键作用，IL2诱导STAT5与IL9启动子和IL-2和IL-21结合，而IL-2和IL-21基于IL-21诱导BCL6，但IL-2的抑制作用在Th9中具有相反的作用。 With Dr. K. Christopher Garcia (Stanford), we previously studied novel IL-2 variant partial agonists that function as "receptor signaling clamps. We previously showed that one variant, H9-RETR, prolonged survival in a model of graft-versus-host disease and blocked spontaneous proliferation of smoldering ATL cells. This receptor-clamping approach might be a general mechanism-based strategy. IL-2 REH was reported as a部分激动剂是特异性扩展的TREG而不是CD8 T细胞，在DSS结肠炎中受益，我们研究了另一种促进CD8 T细胞的TCSM表型的部分激动剂，并在B16黑色素瘤的生育转移治疗中具有增强的疗效，并在第二代抗体模型中使用了Stater 5。导致TSCM表型并阐明了过去一年的基本机制。 IL-21对T-和B细胞具有广泛的作用，我们先前报道说，它还诱导了常规树突状细胞的凋亡。 CHIP-SEQ分析揭示了GM-CSF诱导的STAT5和IL-21诱导的STAT3之间的全基因组结合竞争，我们以前阐明了IL-21信号INT细胞中STAT1与STAT3的作用。 Previously, we also showed that IL-21 regulates expression of the Prdm1 gene that encodes BLIMP1 via a response element that depends on STAT3 and IRF4 and subsequently discovered that in contrast to its known ability to cooperate with PU.1 in B cells to act via Ets-IRF composite elements (EICEs), IRF4 cooperates with BATF/JUN family proteins to act via novel AP1-IRF composite elements (AICEs) in T细胞以及B细胞中。在其他研究中，我们先前研究了EGR1和EGR2的生物学作用，并阐明了EGR1的某些非免疫学作用，表明该转录因子对眼睑发育具有遗传背景依赖性效应 - 在BALB/C背景上进行此类发展所必需，但在C57BL/6背景上不存在。在合作研究中，我们对EGR家族蛋白进行了高级研究，今年发表了一项关于EGR家族蛋白在CNS中Th17细胞在归因和致病性中的作用的合作研究，我们还与吉姆·萨尔泽（Jim Salzer）与EGR蛋白在CNS中的作用有关。 Previously, we studied the biological significance of STAT5 tetramerization in vivo by generating mice expressing mutant forms of STAT5A and STAT5B that could form dimers but not tetramers and previously reported modeling of the 3-dimensional structure of the tetramer, and we also previously reported the basis for defective NK cell development in the double knockin mice, demonstrating a critical role for STAT5 tetramers for the survival of NK cells.在当年，我们现在扩展了对STAT5四聚体的研究，并报告了它们与单核细胞分化的调节有关的作用。我们还以前还以IL-2激活的STAT5和IL-21激活的STAT3调节的超级增强剂及其与高度诱导的基因的关系，发现IL2RA基因包含最排名最高的STAT5依赖性超级增强子。使用CRISPR-CAS9技术，我们以前在功能上剖析了该超级增强剂的元素，为尤其是IL2RA的分子调节提供了关键的新见解，并且更广泛地提供了超级增强剂的分子调节，而在当年，我们已经大大扩展了这些研究，从而阐明了不同的超级元素在不同蜂窝类型中的不同重要性。此外，今年，我们在1型IL-4受体的独特激活中对IL-4模拟物的研究做出了贡献，RNA-Seq表明NEO-4对IL-4和IL-13对单核细胞的影响相似。我们还在一项协作研究中报道了一种有效的新方法Chiatac，用于3D表观基因瘤的多组学映射，并报告了一种新的用户友好的R Shiny应用程序，用于RNA测序分析和生物标志物发现。 总体而言，这些研究增强了我们对GC家族细胞因子调节基因表达和生物学过程的机制的理解，并且与正常和病理免疫细胞功能有关，包括疾病状态，具有潜在的治疗意义。

项目成果

Mitochondrial Membrane Potential Identifies Cells with Enhanced Stemness for Cellular Therapy.

• DOI: 10.1016/j.cmet.2015.11.002
• 发表时间: 2016-01-12
• 期刊: Cell metabolism
• 影响因子: 29
• 作者: Sukumar M;Liu J;Mehta GU;Patel SJ;Roychoudhuri R;Crompton JG;Klebanoff CA;Ji Y;Li P;Yu Z;Whitehill GD;Clever D;Eil RL;Palmer DC;Mitra S;Rao M;Keyvanfar K;Schrump DS;Wang E;Marincola FM;Gattinoni L;Leonard WJ;Muranski P;Finkel T;Restifo NP
• 通讯作者: Restifo NP

BATF-JUN is critical for IRF4-mediated transcription in T cells.

• DOI: 10.1038/nature11530
• 发表时间: 2012-10-25
• 期刊: Nature
• 影响因子: 64.8

Erratum: The transcriptional regulators IRF4, BATF and IL-33 orchestrate development and maintenance of adipose tissue-resident regulatory T cells.

勘误表：转录调节因子 IRF4、BATF 和 IL-33 协调脂肪组织驻留调节性 T 细胞的发育和维持。

• DOI: 10.1038/ni0515-544d
• 发表时间: 2015
• 期刊: Nature immunology
• 影响因子: 30.5
• 作者: Vasanthakumar,Ajithkumar;Moro,Kazuyo;Xin,Annie;Liao,Yang;Gloury,Renee;Kawamoto,Shimpei;Fagarasan,Sidonia;Mielke,LisaA;Afshar-Sterle,Shoukat;Masters,SethL;Nakae,Susumu;Saito,Hirohisa;Wentworth,JohnM;Li,Peng;Liao,Wei;Leonard
• 通讯作者: Leonard

Enhanced T cell lymphoma in NOD.Stat5b transgenic mice is caused by hyperactivation of Stat5b in CD8+ thymocytes.

• DOI: 10.1371/journal.pone.0056600
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Chen B;Yi B;Mao R;Liu H;Wang J;Sharma A;Peiper S;Leonard WJ;She JX
• 通讯作者: She JX

Analysis of interleukin-21-induced Prdm1 gene regulation reveals functional cooperation of STAT3 and IRF4 transcription factors.

• DOI: 10.1016/j.immuni.2009.10.008
• 发表时间: 2009-12-18
• 期刊: Immunity
• 影响因子: 32.4
• 作者: Kwon H;Thierry-Mieg D;Thierry-Mieg J;Kim HP;Oh J;Tunyaplin C;Carotta S;Donovan CE;Goldman ML;Tailor P;Ozato K;Levy DE;Nutt SL;Calame K;Leonard WJ
• 通讯作者: Leonard WJ