Structure, function and engineering of immune cytokine receptor signaling

免疫细胞因子受体信号传导的结构、功能和工程

• 批准号: 10190779
• 负责人: Kenan Christopher GARCIA
• 金额: $ 44.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10190779
• 关键词: Antibodies; Architecture; Award; Binding; Biological; Biological Response Modifiers; Biophysics; Cells; Chemistry; Chemosensitization; Complex; Crystallography; Cytokine Receptors; Disease model; Electron Microscopy; Engineering; Exhibits; Extracellular Structure; Geometry; Goals; Homeostasis; IL6ST gene; Image; Immune; Immune System Diseases; Immune system; Immunotherapeutic agent; Interferons; Interleukin 2 Receptor Gamma; Interleukin-1; Interleukin-13; Interleukin-15; Interleukin-17; Interleukin-2; Interleukin-4; Interleukin-6; Janus kinase; Length; Malignant Neoplasms; Mediating; Molecular; Molecular Conformation; Pharmaceutical Preparations; Play; Privatization; Property; Protein Engineering; Receptor Signaling; Role; STAT protein; Signal Pathway; Signal Transduction; Specificity; Structure; T-Lymphocyte Subsets; Therapeutic; Toxic effect; Variant; X-Ray Crystallography; base; cytokine; immunoengineering; immunoregulation; in vivo; interleukin-15 receptor; interleukin-23; leukocyte proliferation; novel; pleiotropism; preference; receptor; reconstitution

Garcia, K. Christooher PROJECT SUMMARY: lmmunoregulatory cytokines engage transmembrane signaling receptors in order to mediate a wide range of functions including leukocyte proliferation, differentiation, and expansion. Most immunoregulatory cytokines possess both redundant and distinct activities that are critical to normal immune homeostasis, but this functional pleiotropy presents a major problem for the effective, targeted use of these cytokines as drugs. Cytokine pleiotropy is a consequence of a small number of shared receptors, such as common gamma chain and gp130, engaging many different cytokines, which then activate overlapping intracellular signaling pathways through a limited set of JAK and STAT proteins. During the prior term of this award, we gained an appreciation for the extracellular structural architectures of a spectrum of different cytokine complexes with shared receptors, including those of IL-1, IL-2, IL-4, IL-6, IL-13, IL-17, IL-23, and IFN, which exhibited an astonishing diversity of heterodimeric signaling geometries. In this renewal application, we focus our studies on the pleiotropic cytokines IL-2 and IL-15, to ask how extracellular structures of the receptor-cytokine complexes influence transmembrane signaling, intracellular activation of JAK and STAT, and subsequent in vivo function. We wish to determine if the binding chemistry and geometry of the IL-2 and IL-15 receptor complex subunits plays a role in modulating signaling specificity, and whether "tuning" signaling through structure-based cytokine engineering of receptor interactions is a viable means of developing novel immunotherapeutics with enhanced efficacy, cell subset preferences, and reduced toxicity. The overall goals of this highly collaborative proposal are: Aim 1- to determine the biophysical basis for the functional redundancy and specificity exhibited by two "natural surrogate" ye cytokines, IL-2 and IL-15, that act through shared signaling receptors (IL-2Rp and Ye) but private alpha-receptors; Aim 2- to utilize structure-based protein engineering to attempt to create IL-2 variants with diverse signaling properties and T cell subset preferences, that may be more effective immunotherapeutics as assessed by collaborators in a variety of in vivo disease models; and Aim 3- to determine the mechanistic basis of antibody potentiation of IL-2 activity towards distinct T cell subsets, as well as discover new potentiating antibodies that could remodel the conformation of wild-type IL-2 and alter its biological activity. Finally, in Aim 4, we continue to pursue structural information on how cytokine receptor intracellular segments engage Janus Kinase (JAK) molecules, by reconstituting an entire full-length cytokine receptor transmembrane complex, bound to both cytokine and intracellular JAK for imaging by crystallography and electron microscopy. In this fashion, by combining structure (X-ray crystallography, Electron Microscopy, and NMR), protein engineering, signaling, and in vivo studies, we propose to obtain a complete molecular snapshot of shared cytokine receotor sianalina from the initial enaaaement of liaand throuah the activation of intracellular sianalini:i cascades. RELEVANCE: Cytokines are potent mediators of the immune system and they hold promise as therapeutics to treat immune disease and cancer. However, their actions need to be harnessed and controlled in order to enhance the desired effects and minimize undesired effects. This proposal uses protein engineering based on fundamental mechanisms of cytokine-receptor interactions to develop new strategies to control the actions of powerful immune cytokines.

加西亚·K·克里斯托赫 项目概要： 免疫调节细胞因子与跨膜信号受体结合以介导广泛的免疫调节 功能包括白细胞增殖、分化和扩增。大多数免疫调节细胞因子 具有对正常免疫稳态至关重要的冗余和独特的活性，但这种功能 多效性对于有效、有针对性地使用这些细胞因子作为药物提出了一个主要问题。细胞因子 多效性是少数共享受体的结果，例如共同的伽马链和 gp130， 参与许多不同的细胞因子，然后通过a激活重叠的细胞内信号传导途径 有限的 JAK 和 STAT 蛋白集。在该奖项的上一任期内，我们获得了对 具有共享受体的一系列不同细胞因子复合物的细胞外结构体系， 包括IL-1、IL-2、IL-4、IL-6、IL-13、IL-17、IL-23和IFN，它们表现出惊人的多样性 异二聚体信号几何结构。在此更新应用中，我们将研究重点放在多效性细胞因子上 IL-2 和 IL-15，了解受体-细胞因子复合物的细胞外结构如何影响跨膜 信号传导、JAK 和 STAT 的细胞内激活以及随后的体内功能。我们希望确定是否 IL-2 和 IL-15 受体复合物亚基的结合化学和几何形状在调节中发挥作用 信号传导特异性，以及是否通过受体的基于结构的细胞因子工程“调节”信号传导 相互作用是开发具有增强功效、细胞亚群的新型免疫疗法的可行手段 偏好，并降低毒性。这个高度协作提案的总体目标是： 目标 1- 确定 两个“天然替代品”叶表现出的功能冗余和特异性的生物物理学基础 细胞因子 IL-2 和 IL-15，通过共享信号受体（IL-2Rp 和 Ye）但私有 α 受体发挥作用； 目标 2- 利用基于结构的蛋白质工程尝试创建具有不同信号传导的 IL-2 变体 特性和 T 细胞亚群偏好，根据评估，这可能是更有效的免疫治疗 各种体内疾病模型的合作者；目标 3-确定抗体的机制基础 增强 IL-2 对不同 T 细胞亚群的活性，并发现新的增强抗体 可以重塑野生型IL-2的构象并改变其生物活性。最后，在目标 4 中，我们继续 探究细胞因子受体细胞内片段如何参与 Janus 激酶 (JAK) 的结构信息 分子，通过重建整个全长细胞因子受体跨膜复合物，结合到 用于通过晶体学和电子显微镜成像的细胞因子和细胞内 JAK。以这种方式，通过 结合结构（X射线晶体学、电子显微镜和核磁共振）、蛋白质工程、 信号传导和体内研究，我们建议获得共享细胞因子的完整分子快照 受体 sianalini 从最初的 liaaement 和通过细胞内 sianalini:i 级联的激活。 关联： 细胞因子是免疫系统的有效介质，它们有望作为治疗免疫的药物 疾病和癌症。然而，他们的行为需要加以利用和控制，以增强 期望的效果并最大限度地减少不良影响。该提案使用基于基础的蛋白质工程 细胞因子-受体相互作用的机制，以开发新的策略来控制强大的作用 免疫细胞因子。