Immune chemistry and therapeutic features of FOXP3

描述(申请人提供)：这个项目联合会由Mark I.Greene领导，他参与免疫调节和表型逆转的研究已有30多年。这个高度整合的项目的目标是加深对FoxpS蛋白如何发挥其生物学效应的了解，并通过这些知识开发与翻译相关的疗法，使Foxp3复合体失活或激活。虽然Foxp3功能的主题在每个项目中都有共鸣，但实际上使用了各种不同的技术来深入了解Foxp3复合体是如何运作的，以及如何操纵它们，从而产生了一组高度互动的实验，这些实验应该会导致治疗学，并将在该计划项目的时间框架内到达临床。每个项目都重申了长期目标，所有项目都有Treg表型操纵这一反复出现的主题。到目前为止，这些研究已经导致了一种针对自身免疫性疾病的合理治疗方法，并正在进入NIH的初步临床试验。 格林项目的目标是提供有关Foxp3复合体如何与人类细胞中的染色质结合的基本生化信息。这些信息将有助于安德鲁·威尔斯对小鼠染色质-Foxp3相互作用的研究，并将有助于创造转基因和突变小鼠，这将有助于Hancock项目和Wells项目。人类和小鼠的Foxp3复合体似乎有差异，尽管它们都形成了巨大的整体。 项目1的目的是确定乙酰化和磷酸化的单个残基以及调节与其他抑制成分相互作用的亚结构域。这些信息将为安德鲁·威尔斯提供一个框架，以检查小鼠染色质重塑事件，并为韦恩·汉考克提供一个框架，以检查活体模型中的功能相关性。 项目1：FOXP3与免疫调节(格林，M) 项目1说明(由申请人提供)：维持对自身抗原的无反应性对于预防自身免疫是必不可少的，但这是一个不完全了解的过程。我们的研究将集中在调节性T细胞(Treg)的某些生化特征以及Foxp3复合体如何介导其抑制作用。虽然很明显，人类FOXP3的突变使人容易患上人类自身免疫疾病，但为什么突变蛋白无法发挥转录调节作用，目前还不清楚。FOXP3本身如何与转录机制相互作用，以及FOXP3系综中哪些成分产生表型变化以使细胞能够介导抑制的细节也有限。我们的研究集中在FOXP3复合体的生物化学上，以及通过FOXP3整体的合理生化改变来检验Treg功能的体内模型。这些研究提供了令人信服的证据，表明特异组蛋白乙酰转移酶和组蛋白去乙酰基酶(HDAC)的复合体与FOXP3相关，从而产生转录抑制因子。Foxp3被乙酰化和磷酸化，然后能够调节其活动。 赠款的基本领域集中在导致Foxp3整体的翻译后变化和功能的信号效应上。出现的一个主要主题是，Foxp3受到酶的作用，改变其活性和稳定性，并与染色质相互作用，导致Treg在体外和体内的活动发生变化。使用HDAC抑制剂来增加特定赖氨酸残基上的Treg乙酰化修饰，从而增加Treg功能以改善自身免疫，是一种合理的治疗方法。 项目1的目标是提供有关Foxp3复合体如何与人类细胞中的染色质结合的基本生化信息。这些信息将有助于安德鲁·威尔斯对小鼠染色质-FoxpS相互作用的研究，并将有助于创造转基因和突变小鼠，这将有助于Hancock项目和Wells项目。人类和小鼠的Foxp3复合体似乎存在差异。这个项目的目的是确定单个残基是乙酰化和磷酸化的，以及调节与其他抑制成分相互作用的亚域。这些信息将为安德鲁·威尔斯提供一个框架，以检查小鼠染色质重塑事件，并为韦恩·汉考克提供一个框架，以检查活体模型中的功能相关性。

DESCRIPTION (provided by applicant): This confederation of projects is headed by Mark I. Greene who has been involved in the study of immune regulation and phenotype reversal for over 30 years. The goals of this highly integrated program project are to develop understanding of how the FoxpS proteins exert their biological effects and through this knowledge to develop translationally relevant therapeutics that disable or activate the Foxp3 complex. Although the theme of Foxp3 function resonates in each project, a variety of distinct technologies are actually employed to develop insights into how Foxp3 complexes operate and how to manipulate them, leading to a highly interactive group of experiments that should lead to therapeutics that will reach the clinic within the time frame of this program project. The long-term goals are reiterated in every project and all projects share recurrent themes of Treg phenotype manipulation. The studies to date have already lead to a rational therapeutic for autoimmune conditions that is entering a preliminary clinical trial at the NIH. The goal of Greene's project is to provide basic biochemical information of how the Foxp3 complex binds to chromatin in human cells. This information will be helpful in Andrew Wells's study of mouse chromatin - Foxp3 interactions and will be useful in the creation of transgenic and mutant mice that will help the Hancock project and the Wells project. Human and mouse Foxp3 complexes appear to have differences although both form large ensembles. The intent of Project 1 is to identify individual residues that are acetylated and phosphorylated and subdomains that mediate interactions with other repressive components. This information will provide a framework for Andrew Wells to examine chromatin remodeling events in the mouse and for Wayne Hancock to examine functional relevance in in vivo models. PROJECT 1: Foxp3 and immune regulation (Greene, M) PROJECT 1 DESCRIPTION (provided by applicant): Maintenance of unresponsiveness to self-antigens is essential for the prevention of autoimmunity but is an incompletely understood process. Our studies will focus on certain biochemical features of regulatory T cells (Treg) and how the Foxp3 complex mediates its repressive effects. While it is clear that mutations in human FOXP3 predispose individuals to human autoimmune conditions, it is unclear why the mutant protein fails to function as a transcriptional regulator. There is also limited detail of how FOXP3 itself interacts with the transcriptional machinery and which components of the FOXP3 ensembles exert phenotypic changes to render cells able to mediate suppression. Our proposed studies focus on the biochemistry of FOXP3 complexes, as well as in vivo models to examine modification of Treg function through rational biochemical alteration of the FOXP3 ensemble. The studies provide compelling evidence that a complex of specific histone acetyl transferases and histone deacetylases (HDAC) associate with FOXP3 to create a transcriptional represser. FOXP3 becomes acetylated and phosphorylated and is then able to mediate its activities. The essential areas of the grant focus on the signaling effects that lead to post translational changes and functions of the Foxp3 ensemble. A dominant theme that emerges is that Foxp3 is acted on by enzymes that modify its activity and stability and interactions with chromatin which lead to alterations in the actions of Treg in vitro and in vivo. A rational therapeutic emerges from these studies employing HDAC inhibitors to increase Treg acetylation modifications at specific lysine residues and thereby increase Treg function to ameliorate autoimmunity. The goal of Project 1 is to provide basic biochemical information of how the Foxp3 complex binds to chromatin in human cells. This information will be helpful in Andrew Wells's study of mouse chromatin- FoxpS interactions and will be useful in the creation of transgenic and mutant mice that will help the Hancock project and the Wells project. Human and mouse Foxp3 complexes appear to have differences. The intent of this project is to identify individual residues that are acetylated and phosphorylated and subdomains that mediate interactions with other repressive components. This information will provide a framework for Andrew Wells to examine chromatin remodeling events in the mouse and for Wayne Hancock to examine functional relevance in in vivo models.