Diversifying interferon functions through combinatorial and structural biology

描述（由申请人提供）：在这个KO1提案中，我们试图深入探讨分子识别和结构在受体-配体系统中的作用，I型干扰素（IFNs），这对基本受体信号生物学和人类健康具有重要意义。我们打算阐明IFN诱导信号的结构-功能关系，并通过组合生物学、计算生物学和结构生物学来增强IFN的功能。培训和研究计划描述了一个在K. Christopher Garcia博士指导下的五年指导计划，旨在建立Mendoza博士之前的培训，以进一步掌握组合、计算和结构生物学方面的专业知识。这将使Mendoza博士成为一名独立的多学科科学家，并拥有一套强大的工具，使他能够在实验范围内解决癌症生物学细胞因子工程中的问题。我们将尝试了解IFN细胞因子如何通过其对细胞生长、分化和增殖的影响来介导广泛的生理过程。I型干扰素具有抗增殖特性，对恶性细胞的监视和控制很重要。在细胞外受体区域参与后，干扰素在细胞内被不同地激活

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.