Shedding new light on cytokine signaling through molecular engineering

项目总结/摘要 细胞因子信号传导对于启动针对微生物感染和癌症的免疫应答是必不可少的。 免疫系统由两种防御机制组成，即先天性免疫和适应性免疫 这两种系统都严重依赖于细胞因子信号传导来发挥作用。先天免疫系统在早期就开始起作用 在感染或癌症期间，包括I型IFN应答。适应性免疫系统变得完全 大约7天后开始活动。虽然这种反应相对于先天系统是延迟的， 是T细胞产生有效和特异性反应所必需的。白细胞介素-2和干扰素γ是 细胞因子的例子，塑造适应性免疫系统的反应。还有几十个其他的 细胞因子家族，每个家族在免疫系统中起重要作用，包括造血，炎症， 细胞凋亡以及许多其他方面。了解细胞因子如何发出信号，它们诱导的基因， 功能对于了解人类健康和疾病至关重要。工程化细胞因子的最新实例 表明调节细胞因子信号可以彻底改变细胞因子的反应，并可能提供有希望的 新的治疗方法。在这项提案中，我们的目标是利用蛋白质工程技术来填补大规模的 细胞因子信号传导的知识空白，这可能揭示新的靶点和治疗方法 干预 细胞因子信号传导的范例是细胞因子驱动细胞因子受体的二聚化。Janus 激酶（JAK）被认为与细胞因子受体组成性结合。在受体二聚化后， JAK交叉磷酸化彼此以及受体。信号传导与转录活化因子 STAT与磷酸化受体结合，然后磷酸化，二聚化，并易位到细胞核 来引发基因和功能反应。我们实验室最近的工具开发提供了一种简化的方法， 表征细胞内发生的蛋白质-蛋白质相互作用，挑战该领域的假设， 提供了一个机会，了解细胞因子信号传导的每一步如何有助于细胞因子信号传导。我们 旨在显示如何改变这些相互作用调节细胞因子信号和反应基因和功能签名。

Project Summary/Abstract Cytokine signaling is essential to the initiation of the immune response against microbial infection and cancer. The immune system is composed of two mechanisms of defense defined as the innate and adaptive immune systems, both of which critically rely on cytokine signaling to function. The innate immune system acts early during an infection or cancer and includes the type I IFN response. The adaptive immune system becomes fully active after approximately seven days. Although this response is delayed relative to the innate system, the time is needed to mount a T-cell response that is potent and specific. Interleukin-2 and interferon gamma are examples of cytokines that shape the response of the adaptive immune system. There are dozens of other cytokine families that each play an important role in the immune system including hematopoiesis, inflammation, apoptosis as well as many others. Understanding how cytokines signal, the genes they induce, and their functions are critical to understanding human health and disease. Recent examples of engineered cytokines demonstrate that tuning of cytokine signaling can drastically alter a cytokine’s response and may offer promising new therapeutic approaches. In this proposal, we aim to use protein engineering technologies to fill the large gaps in knowledge of cytokine signaling which may reveal new targets and approaches for therapeutic intervention. The paradigm of cytokine signaling is that cytokines drive the dimerization of cytokine receptors. Janus kinases (JAKs) are believed to be constitutively bound to the cytokine receptors. Upon receptor dimerization, the JAKs cross phosphorylate each other as well as the receptors. Signal transducers and activators of transcription (STATs) bind to the phosphorylated receptors, are then phosphorylated, dimerize, and translocate to the nucleus to elicit gene and functional responses. Recent tool development in our lab provides a streamlined approach to characterize protein-protein interactions which occur intracellularly, challenge assumptions in the field, and provide an opportunity to understand how every step in cytokine signaling contributes to cytokine signaling. We aim to show how altering these interactions tune cytokine signaling and response gene and functional signature.