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

项目摘要 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，对艾滋病毒的保护程度各不相同，但没有有效的疫苗可用于大规模人群 呢临床前和临床数据证明了佐剂和抗原对于有效的HIV疫苗的重要性。 疫苗特别地，刺激toll样受体（TLR）或干扰素基因刺激物（STING）可以增强细胞内的免疫应答。 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的 体液免疫和细胞介导免疫的因子，如广泛中和抗体（bnAb） 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， 免疫原递送和佐剂选择。为了克服这些挑战，我们建议将佐剂 衍生的纳米颗粒和工程mRNA用于HIV疫苗的发现。在初步研究中，我们开发了 使用TLR或STING激动剂的佐剂衍生的纳米颗粒（ANP），其显示出巨大的有效治疗潜力。 作为疫苗平台的mRNA递送。此外，我们构建了糖基化的HIV免疫原， 甘露糖结合凝集素（MBL）介导的先天免疫识别，导致增强的抗体应答。 此外，我们系统地研究了mRNA的非翻译区（UTR），以增强 蛋白质生产通过对内源性基因表达的综合分析和从头设计， 为了确定5'和3' UTR的最佳组合，我们鉴定了用于mRNA工程化的5'和3' UTR的最佳组合。基于这些结果 和研究结果，这个项目的目标是开发佐剂衍生的纳米颗粒作为功能性的， 纳米材料能够在体内有效地传递HIV免疫原，从而产生强有力的， 持久的体液和细胞介导的抗HIV免疫力。将实现以下具体目标， 完成我们的目标：1）佐剂衍生的纳米颗粒（ANP）的合成和表征; 2）工程 具有高翻译效率的编码各种HIV免疫原的mRNA转录物的测定;以及3） 小鼠和非人灵长类动物模型中ANPs-mRNA的免疫原性和安全性概况。鼓舞 根据我们初步研究的结果，我们预计这项提案中新设计的纳米材料将建立 这是一种候选疫苗，可以促进临床转化，并为艾滋病毒疫苗的发现开辟了新的途径。 从这项研究中获得的知识也可以扩展到新出现的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.