Purpose: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide. Most current therapeutic strategies primarily include localized treatment, lacking effective systemic strategies. Meanwhile, recent studies have suggested that RNA vaccines can effectively activate antigen-presenting cells (APCs) and lymphocytes to produce a strong systemic immune response and inhibit tumor growth. However, tumor vaccines loaded with a single tumor antigen may induce immunosuppression and immune evasion, while identifying tumor-specific antigens can require expensive and laborious procedures. Therefore, the use of whole tumor cell antigens are currently considered to be promising, potentially effective, methods. Previously, we developed a targeted liposome-polycation-DNA (LPD) complex nanoparticle that possess a small size, high RNA encapsulation efficiency, and superior serum stability. These particles were found to successfully deliver RNA to tumor sites. In the current study, we encapsulated total tumor-derived RNA in lipid nanoparticles (LNPs) to target dendritic cells (DCs) to incite expeditious and robust anti-tumor immunity.
Methods: Total tumor-derived RNA was extracted from liver cancer cells (Hepa1-6 cells). LNPs loaded with tumor RNA were then prepared thin-film hydration method. The ability of RNA LNPs to induce DC maturation, cytotoxicity, and anti-tumor activity, was investigated in vitro and in vivo.
Results: The average particle size of LNPs and RNA LNPs was 102.22 ± 4.05 nm and 209.68 ± 6.14 nm, respectively, while the zeta potential was 29.97 ± 0.61 mV and 42.03 ± 0.42 mV, respectively. Both LNPs and RNA LNP vaccines exhibited good distribution and stability. In vitro, RNA LNP vaccines were capable of promoting DC maturation and inducing T lymphocytes to kill Hepa1-6 cells. In vivo, RNA LNP vaccines effectively prevent and inhibit HCC growth.
Conclusion: RNA LNPs may serve as an effective antigen specific vaccine to induce anti-tumor immunity for HCC.

Keywords: RNA lipid nanoparticles, tumor vaccine, dendritic cells, cancer immunotherapy


中文翻译：

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肿瘤 RNA 脂质纳米颗粒抗肝细胞癌的免疫治疗


目的：肝细胞癌（HCC）是全球癌症相关死亡的主要原因之一。目前的治疗策略大多以局部治疗为主，缺乏有效的全身策略。同时，最近的研究表明，RNA疫苗可以有效激活抗原呈递细胞（APC）和淋巴细胞，产生强烈的全身免疫反应，抑制肿瘤生长。然而，装载单一肿瘤抗原的肿瘤疫苗可能会引起免疫抑制和免疫逃避，而识别肿瘤特异性抗原可能需要昂贵且费力的程序。因此，使用全肿瘤细胞抗原目前被认为是有前途的、潜在有效的方法。此前，我们开发了一种靶向脂质体-聚阳离子-DNA（LPD）复合纳米粒子，该纳米粒子具有小尺寸、高RNA封装效率和优异的血清稳定性。这些颗粒被发现能够成功地将 RNA 递送到肿瘤部位。在当前的研究中，我们将肿瘤源性总 RNA 封装在脂质纳米颗粒 (LNP) 中，以靶向树突状细胞 (DC)，从而激发快速而强大的抗肿瘤免疫。

方法：从肝癌细胞（Hepa1-6细胞）中提取肿瘤来源的总RNA。然后通过薄膜水合方法制备负载肿瘤RNA的LNP。在体外和体内研究了 RNA LNP 诱导 DC 成熟、细胞毒性和抗肿瘤活性的能力。

结果： LNPs和RNA LNPs的平均粒径分别为102.22±4.05 nm和209.68±6.14 nm，zeta电位分别为29.97±0.61 mV和42.03±0.42 mV。 LNP 和 RNA LNP 疫苗均表现出良好的分布和稳定性。在体外，RNA LNP疫苗能够促进DC成熟并诱导T淋巴细胞杀死Hepa1-6细胞。在体内，RNA LNP 疫苗可有效预防和抑制 HCC 生长。

结论： RNA LNPs 可能作为一种有效的抗原特异性疫苗来诱导 HCC 抗肿瘤免疫。


关键词: RNA脂质纳米粒, 肿瘤疫苗, 树突状细胞, 癌症免疫治疗