Do Hai Doan, Marie Corinne, Bessoles Stéphanie, Dhotel Hélène, Seguin Johanne, Larrat Benoit, Doan Bich-Thuy, Scherman Daniel, Escriou Virginie, Hacein-Bey-Abina Salima, Mignet Nathalie
Université de Paris Cité, CNRS, INSERM, UTCBS, 75006 Paris, France.
Chimie ParisTech, Université PSL, F-75005 Paris, France.
Explor Target Antitumor Ther. 2022;3(6):398-413. doi: 10.37349/etat.2022.00090. Epub 2022 Jun 29.
Gene-based immunotherapy against cancer is limited by low gene transfer efficiency. In the literature, interleukin-12 (IL-12) encoding plasmid associated with sonoporation has been shown to enhance antitumoral activity. Moreover, non-viral carriers and high-frequency ultrasound have both been shown to promote immune response activation. Here, IL-12 encoding plasmid, non-viral carrier stimulating the immune response and focused ultrasound were combined in order to improve the antitumoral efficiency.
In order to enhance a gene-based antitumoral immune response, home-made lipids Toll-like receptor 2 (TLR2) agonists and plasmid free of antibiotic resistance version 4 (pFAR4), a mini-plasmid, encoding the IL-12 cytokine were combined with high-intensity focused ultrasound (HIFU). The lipid composition and the combination conditions were selected following and preliminary studies. The expression of IL-12 from our plasmid construct was measured and . The combination strategy was evaluated in mice bearing colon carcinoma cells (CT26) tumors following their weight, tumor volume, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) levels in the serum and produced by splenocytes exposed to CT26 tumor cells.
Lipid-mediated cell transfection and intratumoral injection into CT26 tumor mice using pFAR4-IL-12 led to the secretion of the IL-12 cytokine into cell supernatant and mice sera, respectively. Conditions of thermal deposition using HIFU were optimized. The plasmid encoding pFAR4-IL-12 or TLR2 agonist alone had no impact on tumor growth compared with control mice, whereas the complete treatment consisting of pFAR4-IL-12, TLR2 lipid agonist, and HIFU limited tumor growth. Moreover, only the complete treatment increased significantly mice survival and provided an abscopal effect on a metastatic CT26 model.
The HIFU condition was highly efficient to stop tumor growth. The combined therapy was the most efficient in terms of IL-12 and IFN-γ production and mice survival. The study showed the feasibility and the limits of this combined therapy which has the potential to be improved.
基于基因的癌症免疫疗法受到基因转移效率低下的限制。在文献中，与声穿孔相关的编码白细胞介素-12（IL-12）的质粒已被证明可增强抗肿瘤活性。此外，非病毒载体和高频超声均已被证明可促进免疫反应激活。在此，将编码IL-12的质粒、刺激免疫反应的非病毒载体和聚焦超声相结合，以提高抗肿瘤效率。
为增强基于基因的抗肿瘤免疫反应，将自制的脂质Toll样受体2（TLR2）激动剂和无抗生素抗性版本4（pFAR4）的质粒（一种微型质粒，编码IL-12细胞因子）与高强度聚焦超声（HIFU）相结合。脂质组成和组合条件根据前期研究进行选择。对我们构建的质粒中IL-12的表达进行了检测。在携带结肠癌细胞（CT26）肿瘤的小鼠中，根据其体重、肿瘤体积、血清中干扰素-γ（IFN-γ）和肿瘤坏死因子-α（TNF-α）水平以及暴露于CT26肿瘤细胞的脾细胞产生的这些因子水平，对组合策略进行了评估。
使用pFAR4-IL-12进行脂质介导的细胞转染并瘤内注射到CT26肿瘤小鼠体内，分别导致IL-12细胞因子分泌到细胞上清液和小鼠血清中。优化了使用HIFU的热沉积条件。与对照小鼠相比，单独使用编码pFAR4-IL-12或TLR2激动剂的质粒对肿瘤生长没有影响，而由pFAR4-IL-12、TLR2脂质激动剂和HIFU组成的完整治疗可限制肿瘤生长。此外，只有完整治疗显著提高了小鼠存活率，并对转移性CT26模型产生了远隔效应。
HIFU条件对阻止肿瘤生长非常有效。联合治疗在IL-12和IFN-γ产生以及小鼠存活方面最为有效。该研究表明了这种联合治疗的可行性和局限性，其仍有改进的潜力。
