Khazaei Monfared Yousef, Mahmoudian Mohammad, Zakeri-Milani Parvin, Cecone Claudio, Hayashi Tomoya, Ishii Ken J, Conde João, Matencio Adrián, Trotta Francesco
Department of Chemistry, University of Turin, 10125 Turin, Italy.
Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz 5165665931, Iran.
Cancers (Basel). 2023 Jul 24;15(14):3748. doi: 10.3390/cancers15143748.
mRNA technology has demonstrated potential for use as an effective cancer immunotherapy. However, inefficient in vivo mRNA delivery and the requirements for immune co-stimulation present major hurdles to achieving anti-tumour therapeutic efficacy. Therefore, we used a cationic hyper-branched cyclodextrin-based polymer to increase mRNA delivery in both in vitro and in vivo melanoma cancer. We found that the transfection efficacy of the mRNA-EGFP-loaded Ppoly system was significantly higher than that of lipofectamine and free mRNA in both 2D and 3D melanoma cancer cells; also, this delivery system did not show cytotoxicity. In addition, the biodistribution results revealed time-dependent and significantly higher mEGFP expression in complexes with Ppoly compared to free mRNA. We then checked the anti-tumour effect of intratumourally injected free mRNA-OVA, a foreign antigen, and loaded Ppoly; the results showed a considerable decrease in both tumour size and weight in the group treated with OVA-mRNA in loaded Ppoly compared to other formulations with an efficient adaptive immune response by dramatically increasing most leukocyte subtypes and OVA-specific CD8+ T cells in both the spleen and tumour tissues. Collectively, our findings suggest that the local delivery of cationic cyclodextrin-based polymer complexes containing foreign mRNA antigens might be a good and reliable concept for cancer immunotherapy.
信使核糖核酸（mRNA）技术已显示出作为一种有效的癌症免疫疗法的潜力。然而，体内mRNA递送效率低下以及免疫共刺激的需求是实现抗肿瘤治疗效果的主要障碍。因此，我们使用了一种基于阳离子超支化环糊精的聚合物来提高体外和体内黑色素瘤癌症中的mRNA递送。我们发现，在二维和三维黑色素瘤癌细胞中，负载mRNA-EGFP的Ppoly系统的转染效率显著高于脂质体转染试剂和游离mRNA；此外，该递送系统未显示出细胞毒性。此外，生物分布结果显示，与游离mRNA相比，与Ppoly形成的复合物中mEGFP表达呈时间依赖性且显著更高。然后，我们检查了瘤内注射的游离mRNA-OVA（一种外来抗原）和负载Ppoly的抗肿瘤效果；结果显示，与其他制剂相比，负载Ppoly的OVA-mRNA治疗组的肿瘤大小和重量均显著降低，通过显著增加脾脏和肿瘤组织中的大多数白细胞亚型和OVA特异性CD8+T细胞，产生了有效的适应性免疫反应。总的来说，我们的研究结果表明，局部递送含有外来mRNA抗原的基于阳离子环糊精的聚合物复合物可能是癌症免疫治疗的一个良好且可靠的概念。
