Integration of adjuvant derived nanoparticles and engineered mRNA for HIV vaccine discovery

项目总结 疫苗是阻止艾滋病毒感染传播的一种有希望的方法。虽然疫苗方案在临床上 试验表明，对艾滋病病毒有不同程度的保护，目前还没有有效的疫苗可用于大量人口 现在还不行。临床前和临床数据证明佐剂和抗原对于有效的艾滋病毒的重要性 疫苗。特别是，Toll样受体(TLRs)或干扰素基因刺激物(STING)的刺激增强 HIV抗原在多种动物模型中的免疫应答。与此同时，艾滋病毒免疫原是一个关键 体液免疫和细胞免疫的因子，如广谱中和抗体(BNAbs) 和细胞毒性T细胞。尽管取得了这些重要进展，但在免疫原设计方面仍然存在重大挑战， 免疫基因递送，以及佐剂选择。为了克服这些挑战，我们建议将佐剂 用于HIV疫苗发现的衍生纳米颗粒和工程mRNA。在初步研究中，我们开发了 使用TLR或STING激动剂的佐剂衍生纳米粒(ANP)，显示出高效的巨大潜力 作为疫苗平台的信使核糖核酸传递。此外，我们构建了糖基化的艾滋病毒免疫原，可以触发 甘露糖结合凝集素(MBL)介导的天然免疫识别，导致增强抗体反应。 此外，我们系统地研究了mRNAs的非翻译区(UTRs)，以增强 蛋白质生产。通过对内源基因表达的综合分析和从头设计 在此基础上，我们确定了5‘和3’非编码区的最佳组合，可用于mRNA工程。基于这些结果 和发现，这项拟议项目的目标是开发具有功能性的佐剂衍生纳米颗粒 纳米材料能够有效地在体内传递艾滋病毒免疫原，从而产生强大的和 针对艾滋病毒的持久体液免疫和细胞免疫。将实现以下具体目标 完成我们的目标：1)佐剂衍生纳米粒(ANPS)的合成和表征；2)工程 高翻译效率的编码各种HIV免疫原的mRNA转录本；以及3)检测 ANPS-mRNA在小鼠和非人灵长类动物模型中的免疫原性和安全性。受到以下激励 从我们的初步研究结果，我们预计从这项提案中新设计的纳米材料将建立 一种候选疫苗，这可以促进临床翻译，并为艾滋病毒疫苗发现开辟了一条新的途径。 从这项研究中获得的知识也可以扩展到针对新出现病原体的其他类型的疫苗。

PROJECT SUMMARY A vaccine is a promising approach for stopping the spread of HIV infections. Although vaccine regimens in clinical trials show various levels of protection against HIV, there is no effective vaccine available for a large population yet. Preclinical and clinical data demonstrate the importance of both adjuvants and antigens for an effective HIV vaccine. Particularly, stimulation of toll-like receptors (TLRs) or stimulator of interferon genes (STING) boosts the immune response of the HIV antigens in multiple animal models. Meanwhile, HIV immunogens are a critical factor for both humoral immunity and cell-mediated immunity such as broadly neutralizing antibodies (bnAbs) and cytotoxic T cells. Despite these important advances, significant challenges remain in immunogen design, immunogen delivery, and adjuvant choice. To overcome these challenges, we propose to integrate adjuvant derived nanoparticles and engineered mRNA for HIV vaccine discovery. In preliminary studies, we developed adjuvant derived nanoparticles (ANPs) using TLR or STING agonists, which showed great potential for efficient mRNA delivery as a vaccine platform. Moreover, we constructed glycosylated HIV immunogens that trigger mannose-binding lectin (MBL)-mediated innate immune recognition, leading to enhanced antibody responses. Additionally, we systematically investigated the untranslated regions (UTRs) of mRNAs in order to enhance protein production. Through a comprehensive analysis of endogenous gene expression and de novo design of UTRs, we identified the optimal combination of 5’ and 3’ UTR for mRNA engineering. Based on these results and findings, the goal of this proposed project is to develop adjuvant derived nanoparticles as functional nanomaterials capable of efficiently delivering HIV immunogens in vivo, consequently generating strong and durable humoral and cell-mediated immunity against HIV. The following specific aims will be carried out to accomplish our goal: 1) Synthesis and characterization of adjuvant derived nanoparticles (ANPs); 2) Engineering of mRNA transcripts encoding various HIV immunogens with high translation efficiency; and 3) Determination of immunogenicity and safety profiles of ANPs-mRNA in mouse and non-human primate models. Encouraged by results from our preliminary studies, we expect the newly designed nanomaterials from this proposal to establish a vaccine candidate, which can facilitate clinical translation and a new avenue for HIV vaccine discovery. Knowledge gained from this study can also be extended to other types of vaccines for emerging pathogens.