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: 10262668
• 负责人: Warren J Leonard
• 金额: $ 169.38万
• 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/10262668
• 关键词: Adult T-Cell Leukemia/Lymphoma; Affinity; Agonist; Allergic; Apoptosis; Attenuated; Autoimmune Diseases; B-Cell Development; B-Lymphocytes; BCL6 gene; Binding; Biological; Biological Models; Biological Process; Biology; CD27 Antigens; CD8-Positive T-Lymphocytes; CRISPR/Cas technology; Cell Differentiation process; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Interaction Analysis by Paired-End Tag Sequencing; Closure by clamp; Clustered Regularly Interspaced Short Palindromic Repeats; 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 Transcription; Genomic approach; Granulocyte-Macrophage Colony-Stimulating Factor; Helicobacter Infections; Helminths; Helper-Inducer T-Lymphocyte; Heterodimerization; Host Defense; Human; Human T-lymphotropic virus 1; IL2RA gene; IL6ST gene; IRF4 gene; Immune; Immune response; Immunologic Deficiency Syndromes; Immunologics; Inbred BALB C Mice; Inflammatory; Inflammatory Bowel Diseases; Innate Immune Response; Interferon Type I; Interleukin 2 Receptor; Interleukin 2 Receptor Gamma; Interleukin 4 Receptor; Interleukin-12; Interleukin-15; Interleukin-2; Interleukin-4; Interleukin-7; Interleukin-9; Knock-in Mouse; Malignant - descriptor; Malignant Neoplasms; Methodology; Modeling; Molecular; Mus; Mutate; Natural Killer Cells; PRDM1 gene; Pathologic; Phenotype; Population; Proto-Oncogene Proteins c-jun; Psoriasis; Receptor Signaling; Regulation; Reporting; Repression; Response Elements; Role; STAT protein; STAT1 gene; STAT3 gene; STAT5B gene; Signal Transduction; Staphylococcus aureus; Stat5 protein; Structure of germinal center of lymph node; System; T cell differentiation; T-Cell Activation; T-Lymphocyte; Techniques; Time; Transcription Factor AP-1; Transgenic Mice; Variant; Virus; X-Linked Severe Combined Immunodeficiency; base; cell type; chromatin immunoprecipitation; cytokine; dimer; genome-wide; graft vs host disease; human disease; immune system function; in vivo; insight; interleukin-21 receptor; jun Oncogene; mutant; neutrophil; next generation; novel; pathogen; promoter; protective factors; protein activation; protein expression; receptor; receptor expression; response; three dimensional structure; transcription factor; tumor

The IL-2 receptor and related cytokine/cytokine receptor systems are being studied to understand the T cell immune response in normal and pathologic states. After T-cell activation, the magnitude and duration of the response is controlled in part 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 being 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 intracelllular pathogens, Th2 differentiation vital in allergic disorders/helminths, and Th17 differentiation vital in inflammatory disorders, including psoriasis and inflammatory bowel disease. 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 of cells for Th2 differentiation. Moreover, using genome-wide chromatin immunoprecipitation coupled to DNA sequencing (ChIP-Seq) analysis, we previously found broad regulation of Th2 differentiation via STAT5A and STAT5B and extended these findings by showing that IL-2 via STAT5 induces IL-12Rb2, which is critical for Th1 differentiation. We also showed that IL-2 via STAT5 also regulates T-bet. Interestingly, IL-2 also inhibits expression of IL-6Ra and gp130, helping to explain the inhibition of Th17 differentiation. We also had reported a critical role of IL-2 in Th9 differentiation, with a direct effect of IL-2 on Th9 differentiation via its induction of STAT5 binding to the Il9 promoter and that IL-2 and IL-21 had opposing actions in Th9 differentiation based on induction of BCL6 by IL-21 but repression by IL-2. In the current year, we have studied the role of new molecules, identified by a computational genomics approach, in Th differentiation. We previously collaborated with Dr. K. Christopher Garcia (Stanford), generating novel IL-2 variants, which represent the first partial agonists for a type 1 cytokine. These next-generation IL-2 variants function as "receptor signaling clamps," retaining high affinity for IL-2Rb, inhibiting binding of endogenous IL-2, but their interaction with gc was weakened, attenuating IL-2Rb/gc heterodimerization. We previously showed that one variant, denoted H9-RETR, could prolong survival in a model of graft-versus-host disease and blocked spontaneous proliferation of smoldering adult T cell leukemia (ATL) T cells. This receptor-clamping approach might be a general mechanism-based strategy with applications to other type 1 cytokines as well. During the past year, we continued our study of these molecules and additionally have studied a new IL-2 partial agonist. IL-21 has broad actions on T- and B-cells, and we previously reported that it also induces apoptosis of conventional dendritic cells (cDCs) via STAT3 and Bim, and that this is inhibited by granulocyte-macrophage colony-stimulating factor (GM-CSF). 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 demonstrated 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. Studies of AICEs and IRF4/BATF/JUN/STAT3 were continued, and we also had studied the expression of IRF8 as a protective factor for H. Pylori infection with H.C. Morse and contributed to a study by Axel Kallies showing that IL-2 and IL-12 together with BLIMP1 and TBET control effector CD8 T cell differentiation. In other studies, we had 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 the current year, we extended our studies with H.C. Morse, reporting that the transcription factors IRF8 and PU.1 are required for follicular B cell development and BCL6-driven germinal center responses. We also further identified and reported a role for IL-21 in neutrophil biology and elucidated mechanisms related to its cooperative interplay with type I interferon in regulating the innate immune response to S. aureus. Moreover, in tumor model systems, we elucidated mechanisms underlying actions of IL-2 versus IL-21. 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 intricate 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 tetramers within other cell types as well. 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 ChIA-PET methodology, we had defined long-distance chromatin interactions and by using CRISPR-Cas9 technology, we 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. In the current year, we have significantly extended these studies. We are currently using CRISPR and interaction techniques to further interrogate cytokine systems. 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.

正在研究IL-2受体和相关的细胞因子/细胞因子受体系统，以了解正常和病理状态下的T细胞免疫反应。 T细胞激活后，响应的幅度和持续时间部分由产生的IL-2，IL-2受体的水平及其诱导时间过程控制。 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，在转录水平下进行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分化在炎症性疾病中至关重要，包括牛皮癣和炎症性肠道疾病。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，并控制细胞启动Th2分化。此外，使用与DNA测序（CHIP-SEQ）分析耦合的全基因组染色质免疫沉淀，我们先前通过STAT5A和STAT5B对TH2分化进行了广泛调节，并通过STAT5通过STAT5诱导IL-12RB2来扩展这些发现，这对于TH1分化至关重要。我们还表明，IL-2通过STAT5还调节T-BET。有趣的是，IL-2还抑制了IL-6RA和GP130的表达，有助于解释对Th17分化的抑制作用。我们还报道了IL-2在Th9分化中的关键作用，IL-2通过诱导STAT5与IL9启动子结合而对Th9分化的直接作用，并且IL-2和IL-21在IL-21诱导BCL6基于IL-21的诱导基于IL-21的诱导中具有相反的作用，但由IL-21抑制。在当年，我们研究了通过计算基因组学方法在分化中识别的新分子的作用。 我们以前与K. Christopher Garcia博士（斯坦福大学）合作，生成了新颖的IL-2变体，这代表了1型细胞因子的第一个部分激动剂。这些下一代IL-2变体充当“受体信号夹”，保留了对IL-2RB的高亲和力，抑制了内源性IL-2的结合，但它们与GC的相互作用削弱了，削弱了IL-2RB/GC异二何数。我们先前表明，一种称为H9-RETR的变体可以在移植物抗宿主疾病模型中延长生存，并阻塞闷烧成人T细胞白血病（ATL）T细胞的自发增殖。这种受体钳方法可能是基于一般机制的策略，其应用于其他1型细胞因子。在过去的一年中，我们继续研究这些分子，并研究了一种新的IL-2部分激动剂。 IL-21对T-和B细胞具有广泛的作用，我们先前报道说，它还通过STAT3和BIM诱导常规树突状细胞（CDC）的凋亡，并且这受到粒细胞 - 巨噬细胞刺激性刺激因子（GM-CSF）的抑制。 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。以前，我们还证明了IL-21调节PRDM1基因的表达，该基因通过响应元素编码Blimp1的pRDM1基因的表达，该响应元素取决于STAT3和IRF4，随后发现，与其在B细胞中与PU.1合作的能力相反，它可以通过ETS-IRF复合元素（EECES）与irf4合作（irf4 compatire in ap compatire in Native in Notive in compatir in compatire in Native in Nothice Intir in Native in Nothice Inter in in Native in in Nothice Inter）（ENSIRE）（ENSIRE in Native Interir in Nothical Inter）（ T细胞以及B细胞中。对AICS和IRF4/BATF/JUN/STAT3的研究进行了研究，我们还研究了IRF8作为H.C.幽门螺杆菌感染的保护因素的表达。莫尔斯（Morse）并促进了Axel Kallies的一项研究，表明IL-2和IL-12与Blimp1和TBET控制效应子CD8 T细胞分化。在其他研究中，我们研究了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背景上进行这种发展所必需的，但在C57BL/6背景上不存在。在当年，我们扩展了H.C.的研究。莫尔斯（Morse）报告说，卵泡B细胞发育和Bcl6驱动的生发中心反应所必需的转录因子IRF8和PU.1。我们还进一步鉴定并报告了IL-21在中性粒细胞生物学中的作用，并阐明了与其与I型干扰素的合作相互作用相关的机制在调节对金黄色葡萄球菌的先天免疫反应中的作用。此外，在肿瘤模型系统中，我们阐明了IL-2对IL-21的作用的基础机制。 以前，我们通过生成表达STAT5A和Stat5b突变形式的小鼠在体内研究的生物学意义，这些小鼠可能形成二聚体，但不建议形成四聚体，并在四维结构中进行了四维结构的复杂建模，并且以前还报告了Douptial nockin nk commite nk Cells nk commite complief neckin nk commite 5的基础，以表现为Douptial nockin nem neckin complatival neckin complative tat comptival nective neckin sevival in Stat complative nect in Station 5 sating 5 nctect n.在当年，我们现在也扩展了对其他细胞类型中四聚体的研究。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CHIA-PET方法，我们定义了长距离染色质相互作用并使用CRISPR-CAS9技术，我们在功能上剖析了该超级增强剂的元素，为IL2RA的分子调节提供了关键的新见解，特别是超级增强剂。在当年，我们大大扩展了这些研究。我们目前正在使用CRISPR和交互技术来进一步询问细胞因子系统。 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。