理据：虽然基于新抗原的癌症疫苗在各种实体肿瘤中显示出潜力，但晚期疾病患者的免疫反应和临床疗效有限。细胞质内运输新抗原和佐剂是激活细胞内Toll样受体（TLRs）和交叉呈递以激活新抗原特异性CD8 + T细胞所必需的，但仍然是一个很大的挑战。方法：在本研究中，我们旨在开发一种具有独特的刺突拓扑结构的病毒样硅疫苗（V-scVLPs），能够高效共运载肝细胞癌（HCC）特异性新抗原和TLR9激动剂到树突状细胞（DCs），以诱导强有力的CD8 + T细胞应答，以预防原位肿瘤生长。我们通过检查动物模型中的肿瘤生长和生存时间，以及分析肿瘤微环境中的肿瘤浸润CD8 + T细胞和细胞因子应答来评估V-scVLPs的抗肿瘤疗效。为了评估V-scVLPs与α-TIM-3在HCC中的协同疗效，我们使用了原位HCC小鼠模型，肺转移模型和肝切除后的肿瘤再挑战模型。结果：我们发现，V-scVLPs能够通过穴窖介导的内吞作用高效共运载肝细胞癌（HCC）特异性新抗原和TLR9激动剂到DCs。这种先进的运输策略导致V-scVLPs能够被淋巴结引流，以激活淋巴样DC成熟，促进强有力的CD8 + T细胞和中央记忆T细胞的应答，从而有效预防原位HCC肿瘤生长。然而，在已建立的原位肝肿瘤模型中，抑制性受体TIM-3在接种V-scVLPs后的肿瘤浸润CD8 + T细胞中显著上调。阻断TIM-3信号可以进一步恢复V-scVLPs引发的CD8 + T细胞的抗肿瘤活性，减少调节性T细胞的比例，并增加细胞因子的水平，改变肿瘤微环境，有效抑制已建立的原位HCC肿瘤生长，并抑制肺转移以及肝切除后的复发。结论：总的来说，这种具有高效新抗原和佐剂胞内传递能力的新型刺突纳米颗粒对于未来改善HCC免疫治疗具有巨大的潜力。© 作者。

Rationale: Although neoantigen-based cancer vaccines have shown promise in various solid tumors, limited immune responses and clinical outcomes have been reported in patients with advanced disease. Cytosolic transport of neoantigen and adjuvant is required for the activation of intracellular Toll-like receptors (TLRs) and cross-presentation to prime neoantigen-specific CD8+T cells but remains a significant challenge. Methods: In this study, we aimed to develop a virus-like silicon vaccine (V-scVLPs) with a unique spike topological structure, capable of efficiently co-delivering a hepatocellular carcinoma (HCC)-specific neoantigen and a TLR9 agonist to dendritic cells (DCs) to induce a robust CD8+T cell response to prevent orthotopic tumor growth. We evaluated the antitumor efficacy of V-scVLPs by examining tumor growth and survival time in animal models, as well as analyzing tumor-infiltrating CD8+T cells and cytokine responses in the tumor microenvironment (TME). To evaluate the synergistic efficacy of V-scVLPs in combination with α-TIM-3 in HCC, we used an orthotopic HCC mouse model, a lung metastasis model, and a tumor rechallenge model after hepatectomy. Results: We found that V-scVLPs can efficiently co-deliver the hepatocellular carcinoma (HCC)-specific neoantigen and the TLR9 agonist to DCs via caveolin-mediated endocytosis. This advanced delivery strategy results in efficient lymph node draining of V-scVLPs to activate lymphoid DC maturation for promoting robust CD8+T cells and central memory T cells responses, which effectively prevents orthotopic HCC tumor growth. However, in the established orthotopic liver tumor models, the inhibitory receptor of TIM-3 was significantly upregulated in tumor-infiltrating CD8+T cells after immunization with V-scVLPs. Blocking the TIM-3 signaling further restored the antitumor activity of V-scVLPs-induced CD8+T cells, reduced the proportion of regulatory T cells, and increased the levels of cytokines to alter the tumor microenvironment to efficiently suppress established orthotopic HCC tumor growth, and inhibit lung metastasis as well as recurrence after hepatectomy. Conclusion: Overall, the developed novel spike nanoparticles with efficient neoantigen and adjuvant intracellular delivery capability holds great promise for future clinical translation to improve HCC immunotherapy.© The author(s).