Alshehry Yasir, Liu Xiang, Zhang Yu, Zhu Guizhi
Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, VA 23298, USA; Department of Pharmaceutics, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam 31441, Saudi Arabia.
Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA.
J Control Release. 2025 May 10;381:113617. doi: 10.1016/j.jconrel.2025.113617. Epub 2025 Mar 17.
Circular RNA (circRNA) is an emerging class of vaccines for various diseases, such as cancer immunotherapy. For cancer therapeutic vaccines, it is critical to deliver circRNA to lymphoid tissues such as lymph nodes (LNs) and dendritic cells (DCs) and then elicit antigen-specific T cell responses. Lipid nanoparticles (LNPs) have shown great success for mRNA vaccines and may also have great potential as nanocarriers for circRNA vaccines. Here, we studied the impact of LNP composition on the efficiency of immune delivery, protein expression, and the T cell responses for circRNA vaccine. First, we used model mRNA and circRNA encoding firefly luciferase (mRNA-fLuc) to study protein expression and used two small circRNA vaccines to study T cell responses. We investigated a combination of six ionizable lipids, three helper lipids, and six different molar ratios of cholesterol and β-sitosterol for their impact on the physicochemical properties of RNA LNPs, in vitro DC transfection, in vivo protein expression in draining LNs, and antigen-specific T cell responses. Among these ionizable lipids, SM-102 was the most effective for DC transfection and enabling circRNA vaccines to elicit T cell responses. DOPE and β-sitosterol incorporation in LNPs resulted in efficient protein expression, albeit β-sitosterol incorporation appeared to be associated with reduced T cell response. Overall, circRNA was efficiently delivered to DCs and macrophages in mouse draining lymph nodes by LNPs of SM-102 (50 %), cholesterol (38.5 %), DOPE (10 %), and DMG-PEG2000 (1.5 %), resulting in the induction of potent antigen-specific CD8 T cell response in mice. These findings may provide insights into designing the compositions of LNPs as the carrier for circRNA therapeutics and vaccines.
环状RNA(circRNA)是一类新兴的用于多种疾病的疫苗,如癌症免疫治疗。对于癌症治疗性疫苗而言,将circRNA递送至淋巴结(LNs)和树突状细胞(DCs)等淋巴组织,进而引发抗原特异性T细胞反应至关重要。脂质纳米颗粒(LNPs)在mRNA疫苗方面已取得巨大成功,作为circRNA疫苗的纳米载体也可能具有巨大潜力。在此,我们研究了LNP组成对circRNA疫苗免疫递送效率、蛋白质表达及T细胞反应的影响。首先,我们使用编码萤火虫荧光素酶的模型mRNA和circRNA(mRNA-fLuc)来研究蛋白质表达,并使用两种小型circRNA疫苗来研究T细胞反应。我们研究了六种可电离脂质、三种辅助脂质以及胆固醇与β-谷甾醇六种不同摩尔比的组合对RNA-LNPs物理化学性质、体外DC转染、引流淋巴结中的体内蛋白质表达以及抗原特异性T细胞反应的影响。在这些可电离脂质中,SM-102对DC转染以及使circRNA疫苗引发T细胞反应最为有效。在LNP中掺入DOPE和β-谷甾醇可实现高效的蛋白质表达,尽管掺入β-谷甾醇似乎与T细胞反应降低有关。总体而言,由50%的SM-102、38.5%的胆固醇、10%的DOPE和1.5%的DMG-PEG2000组成的LNP能够将circRNA有效地递送至小鼠引流淋巴结中的DC和巨噬细胞,从而在小鼠体内诱导出强效的抗原特异性CD8 T细胞反应。这些发现可能为设计作为circRNA治疗药物和疫苗载体的LNP组成提供见解。
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