Diversifying interferon functions through combinatorial and structural biology

通过组合和结构生物学使干扰素功能多样化

• 批准号: 9136049
• 负责人: Juan Luis Mendoza
• 金额: $ 11.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-18 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9136049
• 关键词: Affinity; Basic Science; Binding; Biochemistry; Biological Assay; Biological Response Modifier Therapy; Biology; Cancer Biology; Cell membrane; Cells; Complex; Computational Biology; Computer Simulation; Coupled; Cytokine Receptors; Cytokine Signaling; DNA; Development; Dimerization; Directed Molecular Evolution; Disease; Engineering; Equilibrium; Event; Evolution; Exhibits; Family; Flowcharts; Future; Geometry; Goals; Health; Human; Immune; Immune System Diseases; In Vitro; Individual; Interferon Receptor; Interferon Type I; Interferons; Ligand Binding; Ligands; Malignant Neoplasms; Mediating; Mentors; Methodology; Molecular; Molecular Structure; Nature; Output; Physiological Processes; Play; Property; Publishing; Receptor Signaling; Research Proposals; Role; Scientist; Shapes; Signal Pathway; Signal Transduction; Specificity; Structure; Structure-Activity Relationship; System; Techniques; Testing; Therapeutic; Toxic effect; Training; Translating; Variant; Viral; Virus; Virus Diseases; Vision; Work; adaptive immunity; arm; base; cancer cell; cancer therapy; cell growth; combinatorial; cytokine; design; extracellular; improved; insight; member; molecular recognition; novel; programs; receptor; receptor binding; receptor-mediated signaling; research and development; research study; response; structural biology; therapeutic development; tool; tumor

DESCRIPTION (provided by applicant): In this KO1 proposal, we attempt to deeply interrogate the role of molecular recognition and structure in a receptor-ligand system, type I Interferons (IFNs), that has fundamental implications for basic receptor signaling biology as well as human health. We intend to elucidate the structure- functional relationships of IFN induced signaling and to enhance IFN functions through combinatorial, computational, and structural biology. The training and research proposal describes a five year mentored program under the guidance of Dr. K. Christopher Garcia designed to build upon Dr. Mendoza's previous training in order to further expertise in combinatorial, computational, and structural biology. This will establish Dr. Mendoza as an independent multi-disciplinary scientist with a powerful set of tools to give him the experimental range to tackle problems in cytokine engineering for cancer biology. We will attempt to understand how IFN cytokines mediate a wide range of physiological processes through their effects on cell growth, differentiation, and proliferation. Type I IFNs have anti-proliferative properties important for the surveillance and control of malignant cells. Upon engagement of their receptor extracellular regions, IFNs differentially activate intracellular JAK/STAT signaling cascades. Members of the type I IFN family elicit different signaling responses despite binding to a common pair of receptors, IFN?R1 and IFN?R2. Fundamentally, the specificity of cytokine signaling is largely determined by the composition of receptor chains within the signaling complex. While much is known about cytokine-activated intracellular signaling pathways, we know much less about how extracellular structural changes induced by ligand binding translates into signaling events. The experiments proposed will provide more clarity to this question. The potential translational and cancer therapeutic applications are based on a pioneering study published more than ten years ago suggesting that IFN diversification through in vitro evolution could yield molecules with enhanced functional properties. Now, armed with powerful new methodologies, we wish to revisit this concept to create IFNs with diverse functional properties, but also to provide a structural and mechanistic rationale for these altered activities in a way that could inform strategies to engineer IFNs for cancer therapy in the future. We will achieve this by 1) in vitro evolution of IFN?1, 2) identification of variants with altered function using in vitro anti-viral and anti-proliferative assays, 3) structural characterization of the ternary complexes of the evolved IFNs to correlate structure with function, and 4) the computational design and in vitro evolution of chimeric cytokines with distinct and novel signaling and functional outputs. By understanding the molecular basis of cytokine/receptor interactions with correlated function, the results from this work may be able to guide the future development of cytokines as therapeutics as well as expand the capabilities and diversity of receptor mediated signaling, functional activities, and therapeutic properties.

描述（由申请人提供）：在此KO1提案中，我们试图深入询问分子识别和结构在受体配体系统中的作用，I型I干扰素（IFNS）对基本受体信号生物学以及人类健康具有根本性的影响。我们打算阐明IFN诱导的信号传导的结构功能关系，并通过组合，计算和结构生物学增强IFN功能。培训和研究建议描述了在K. Christopher博士的指导下进行了为期五年的指导计划，旨在在Mendoza博士先前的培训上进行培训，以便在组合，计算和结构生物学方面进一步专业知识。这将建立Mendoza博士为独立的多学科科学家，并拥有一套强大的工具，可以为他提供实验范围，以解决癌症生物学的细胞因子工程问题。我们将尝试了解IFN细胞因子如何通过对细胞生长，分化和增殖的影响来介导广泛的生理过程。 I型IFN具有对恶性细胞监视和控制重要的抗增殖特性。在受体细胞外区域互动后，IFN差异激活细胞内 jak/stat信号级联。 I型IFN家族的成员尽管与常见的受体IFN？R1和IFN？r2结合了不同的信号反应。从根本上讲，细胞因子信号的特异性在很大程度上取决于信号传导复合物中受体链的组成。尽管对细胞因子激活的细胞内信号通路知之甚少，但我们对配体结合引起的细胞外结构变化如何转化为信号事件。提出的实验将为这个问题提供更明确的清晰度。潜在的转化和癌症治疗应用是基于十多年前发表的一项开创性研究，表明通过体外进化而多样化可以产生具有增强功能特性的分子。现在，拥有强大的新方法，我们希望重新审视这一概念，以创建具有多种功能性能的IFN，同时也为这些改变的活动提供结构性和机械性原理，以一种可以为未来策略提供策略的方式，以便将来进行癌症治疗。 We will achieve this by 1) in vitro evolution of IFN?1, 2) identification of variants with altered function using in vitro anti-viral and anti-proliferative assays, 3) structural characterization of the ternary complexes of the evolved IFNs to correlate structure with function, and 4) the computational design and in vitro evolution of chimeric cytokines with distinct and novel signaling and functional outputs.通过了解细胞因子/受体相互作用与相关功能的分子基础，这项工作的结果可能能够指导细胞因子作为治疗剂的未来发展，并扩大受体介导的信号传导，功能活性和治疗特性的能力和多样性。