Synthetic biology-regulated RNA vaccines

派：欧文，达雷尔·J。 项目摘要/摘要： 促进免疫后T细胞和抗体反应的数量和质量的策略 开发新的癌症预防和治疗性疫苗具有广泛的相关性 和传染病。最近的研究，包括我们自己实验室的工作，已经证明 抗原和佐剂暴露于淋巴组织的动力学模式对免疫有实质性影响 对接种疫苗的反应。然而，主动控制抗原/炎症线索的时间模式 在目前的所有疫苗方法中，都缺乏对淋巴结的递送。在这里，我们提出了一种应用于 从合成生物学中创造核酸疫苗的方法，其中疫苗抗原/佐剂 表达动态可以通过(I)口服FDA批准的Small的外源调节来控制 分子药物或(Ii)在RNA携带的遗传电路中内在编程。基于前景看好的 针对RNA疫苗的特点，在初步研究中，我们建立了一种脂类纳米粒自递送 复制甲型病毒复制子RNA作为这些受监管疫苗的平台。我们将系统地研究 疫苗抗原和佐剂动力学对接种疫苗的免疫反应的影响，产生前 程序化疫苗动力学模式，并测试受调控复制子的能力，以实现单次免疫 由口服小分子药物控制的优质和增强疫苗。我们的具体目标是(1) 从RNA复制子中优化抗原和分子佐剂的小分子调控表达，(2) 使用调控的复制子平台来确定抗原和佐剂表达的最佳动力学 疫苗接种，(3)设计基于RNA的复制子遗传电路，具有预先编程的时间模式，以及 (4)确定体内限制复制子表达寿命的因素，并设计延长复制子表达寿命的策略 朝着小分子调控的优质增强方案的目标表达。这些研究将导致 基础免疫学的基本发现，为合理设计免疫提供了一个框架 养生方案，并创造技术来实际实施这些方案。

PI: Irvine, Darrell J. Project Summary/Abstract: Strategies to promote the magnitude and quality of T cell and antibody responses following immunization have broad relevance for the development of new prophylactic and therapeutic vaccines for the treatment of cancer and infectious diseases. Recent studies, including work from our own laboratories, have demonstrated that the kinetic pattern of antigen and adjuvant exposure to lymphoid tissues has a substantial impact on the immune response to vaccination. However, active control over the temporal pattern of antigen/inflammatory cue delivery to lymph nodes is lacking in all current vaccine approaches. Here we propose an approach applying methods from synthetic biology to create nucleic acid-based vaccines where vaccine antigen/adjuvant expression dynamics can be controlled by (i) exogenous regulation by orally-available FDA-approved small molecule drugs or (ii) intrinsically programmed in genetic circuits carried by the RNA. Based on the promising features of RNA-based vaccines, in preliminary studies we established a lipid nanoparticle-delivered self- replicating alphavirus replicon RNA as the platform for these regulated vaccines. We will systematically study the impact of vaccine antigen and adjuvant kinetics on the immune response to vaccination, create pre- programmed vaccine kinetic patterns, and test the capacity of regulated replicons to enable single-shot vaccines with prime and boost controlled by an orally-available small molecule drug. Our specific aims are (1) To optimize small molecule-regulated expression of antigen and molecular adjuvants from RNA replicons, (2) To use the regulated replicon platform to define optimal kinetics of antigen and adjuvant expression during vaccination, (3) To design RNA-based replicon genetic circuits with pre-programmed temporal patterns, and (4) To determine factors limiting replicon expression lifetimes in vivo, and engineer strategies to prolong expression toward the goal of small molecule-regulated prime-boost regimens. These studies will lead to fundamental discoveries in basic immunology, provide a framework for rationally designing immunization regimens, and create technologies to practically implement them.