Controlling Vaccine Kinetics with Small Molecule Drugs

项目总结 艾滋病毒、疟疾、结核病和季节性流感流行等传染病和新出现的 新的流行病仍然是主要的全球健康问题，突显了疫苗方面创新方法的必要性 设计。已知与炎症信号相关的抗原暴露的精确动力学在 形成协调的细胞和体液免疫，从而增强疫苗的免疫原性和 功效。然而，目前的疫苗接种战略不包括临时控制抗原的机制。 以及与淋巴组织接触的佐剂。在这里，我们建议将合成生物学方法 创造基于核酸的疫苗，其中疫苗(抗原和佐剂)暴露的动力学可以 使用FDA批准的口服小分子药物进行控制。此策略可通过以下方式启用 自我复制的RNA称为复制子，编码抗原和细胞因子分子佐剂，并包含 由FDA批准的小分子药物甲氧苄啶(TMP)管理的调控机制。这一战略 允许通过单次注射在体内传递编码抗原和细胞因子的复制体，然后 口服TMP控制抗原和细胞因子表达的幅度和持续时间。vbl.使用 RNA复制子平台提供了几个优点：(I)它允许抗原和细胞因子在 它们的天然构象；(Ii)它自我复制，因此在细胞内持续的时间比mRNA和 维持稳定的“新鲜”抗原和佐剂供应；(Iii)与DNA疗法不同，它不存在风险 而且也不需要将转基因表达送到细胞核。一种艾滋病病毒 被称为工程外区(EOD-GT8)的包膜免疫原将被用作模型抗原， 白细胞介素2和白细胞介素12将作为细胞因子分子佐剂。 该项目的具体目标是：(1)产生小分子反应的RNA复制子疫苗，使 控制抗原和佐剂表达的动态。(2)确定抗原的最佳时间模式 以及佐剂暴露，最大限度地提高复制子疫苗引起的保护性免疫力。本项目的成果 将建立一个疫苗平台，允许调节和促进T细胞的数量和质量 通过口服药物进行免疫后的抗体反应，作为一种简单和临床上- 增强疫苗诱导免疫的可翻译策略。此外，阐明最适的细胞因子和 提供保护的抗原暴露模式将为受孕提供关键信息和见解 更好的疫苗设计策略。

PROJECT SUMMARY Infectious diseases such as HIV, malaria, tuberculosis, and seasonal influenza epidemics and emergence of new pandemics remain major global health problems highlighting the need for innovative approaches in vaccine design. The precise kinetics of antigen exposure relative to inflammatory cues is known to play a critical role in shaping a coordinated cellular and humoral immunity and thereby enhancing the vaccine immunogenicity and efficacy. Current vaccination strategies, however, do not include mechanisms for the temporal control of antigen and adjuvant exposure to lymphoid tissues. Here, we propose incorporating synthetic biology approaches to create nucleic acid-based vaccines where the kinetics of vaccine (antigen and adjuvant) exposure can be controlled using orally-available FDA-approved small molecule drugs. This strategy is enabled using self-replicating RNAs termed replicons that encode antigens and cytokine molecular adjuvants and encompass regulatory mechanisms governed by the FDA-approved small molecule drug, trimethoprim (TMP). This strategy allows the delivery of replicons encoding antigens and cytokines in vivo with a single bolus injection and then controlling the amplitude and duration of antigen and cytokine expression by oral administration of TMP. Using the RNA replicon platform provides several advantages: (i) it allows antigens and cytokines to be produced in their native conformation; (ii) it self-replicates and therefore persists inside the cells longer than mRNA and sustains a steady supply of “fresh” antigen and adjuvant; (iii) unlike DNA therapeutics, it does not harbor the risk of genome integration and also does not require delivery to the nucleus for transgene expression. An HIV envelope immunogen, known as the engineered outer domain (eOD-GT8), will be used as the model antigen, and interleukine-2 and interleukine-12 will be used as the cytokine molecular adjuvants. The Specific Aims of this project are: (1) Generate small molecule-responsive RNA replicon vaccines enabling control over the dynamics of antigen and adjuvant expression. (2) Identify optimal temporal patterns of antigen and adjuvant exposure maximizing the protective immunity elicited by replicon vaccines. Results from this project will establish a vaccine platform that allows modulating and promoting the magnitude and quality of T cell and antibody responses following immunization by taking a drug available as an oral pill, as a simple and clinically- translatable strategy to enhance vaccine-induced immunity. In addition, elucidating the optimal cytokine and antigen exposure patterns that confer protection will provide critical information and insights for use in conceiving better vaccine design strategies.