Neoantigen-based personalized vaccination has emerged as a viable method for tumor immunotherapy. Here we set up a DNA-based neoantigen vaccine platform with comprehensive identification of individual somatic mutations using whole-exome sequencing (WES) and RNA-seq, bioinformatic prediction of neo-epitopes, dendritic cell (DC)-based efficacy prevalidation of vaccine candidates, optimization of the DNA vaccine and its nanocarrier and adjuvant, and preparation of a liposome-encapsulated multiepitope DNA vaccine. The DNA vaccine was efficiently uptaken by DCs and induced effective immune response against mouse melanoma cells, leading to significant inhibition of melanoma tumor growth and reduction of lung metastasis in a mouse model. Numerous intratumoral infiltrated CD8+ T-cells with specific in vitro killing ability towards melanoma cells were identified. Our study offers evidence that a multiepitope neoantigen DNA vaccine in a nanocarrier can be exploited for personalized tumor immunotherapy and as a reliable prevalidation approach for rapid enrichment of effective neoantigens.

基于新抗原的个性化疫苗接种已成为一种可行的肿瘤免疫治疗方法。在这里，我们建立了一个基于 DNA 的新抗原疫苗平台，使用全外显子组测序 (WES) 和 RNA-seq 全面识别个体体细胞突变，新表位的生物信息学预测，基于树突状细胞 (DC) 的疫苗候选疗效预验证，优化DNA疫苗及其纳米载体和佐剂，制备脂质体包裹的多表位DNA疫苗。DNA疫苗被DCs有效摄取并诱导针对小鼠黑色素瘤细胞的有效免疫反应，从而显着抑制黑色素瘤肿瘤的生长并减少小鼠模型中的肺转移。大量肿瘤内浸润的 CD8+ T 细胞具有特异性体外确定了对黑色素瘤细胞的杀伤能力。我们的研究提供的证据表明，纳米载体中的多表位新抗原 DNA 疫苗可用于个性化肿瘤免疫治疗，并作为一种可靠的预验证方法，用于快速富集有效的新抗原。