Strong Immune Responses Induced by Direct Local Injections of Modified mRNA-Lipid Nanocomplexes
Strong Immune Responses Induced by Direct Local Injections of Modified mRNA-Lipid Nanocomplexes
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DOI：
10.1016/j.omtn.2019.12.044
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发表时间：
2020-03-06
影响因子：
8.8
通讯作者：
Huang, Jian-Dong
中科院分区：
文献类型：
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作者：
Arya, Smriti;Lin, Qiubin;Huang, Jian-Dong
体外转录mrna在疾病预防和治疗方面具有许多医学应用前景，例如替代或补充缺失或表达不足的内源性蛋白质，以及作为预防传染病的疫苗，治疗性疫苗或用于癌症治疗的其他基于蛋白质的生物制药。一个安全有效的mRNA递送系统是mRNA治疗应用成功的关键。在这项研究中，我们报道了InstantFECT，一种基于脂质体的转染试剂，可以将假尿嘧啶结合的mRNA包装成纳米复合物，在局部递送后高效地介导多器官的体内转染。在瘤内和肌肉注射后，观察到EGFP和荧光素酶报告蛋白的高水平表达，持续时间长达96小时。通过皮下递送编码金黄色葡萄球菌腺苷合酶A （AdsA）和模型肿瘤相关抗原卵清蛋白（OVA）的修饰mRNA，研究了纳米复合物递送mRNA编码抗原的免疫原性。传递的两种mrna都能引起强烈的T细胞反应。OVA mrna -脂质体纳米复合物的治疗性和保护性治疗可显著抑制B16-OVA肿瘤进展并提高小鼠存活率。在组织培养和小鼠实验中均未发现明显的毒性反应。静脉注射相同剂量的修饰mRNA-脂质纳米复合物在主要器官中的转染量最小，表明具有极好的安全性，因为基因转移仅发生在注射部位，而静脉注射相同剂量的mRNA与商业转染试剂Trans-IT复合物在脾脏中显示荧光素酶表达。总之，InstantFECT阳离子脂质体提供了一种安全有效的mRNA在体内局部递送，可以成为基础研究和基于mRNA的治疗开发的有用工具。
In vitro transcribed mRNAs hold the promises of many medical applications in disease prevention and treatment, such as replacement or supplement of missing or inadequately expressed endogenous proteins and as preventive vaccines against infectious diseases, therapeutic vaccines, or other protein-based biopharmaceutics for cancer therapy. A safe and efficient delivery system for mRNA is crucial to the success of mRNA therapeutic applications. In this study, we report that InstantFECT, a liposome-based transfection reagent, can pack pseudouridine-incorporated mRNA into nanocomplexes that are highly efficient in mediating in vivo transfection in multiple organs after local delivery. High levels of expression of EGFP and luciferase reporters after intratumoral and intramuscular injections were observed, which lasted for up to 96 hrs. Immunogenicity of antigens encoded by mRNA delivered with nanocomplex was investigated by subcutaneous delivery of modified mRNAs encoding Staphylococcus aureus adenosine synthase A (AdsA) and a model tumor-associated antigen ovalbumin (OVA). Strong T cell responses were provoked by both mRNAs delivered. Therapeutic and protective treatment with the OVA mRNA-liposome nanocomplex significantly inhibited B16-OVA tumor progression and increased mouse survival. There was no sign of obvious toxicity related to the treatment both in tissue culture and in mice. An intravenous injection of the same dosage of the modified mRNA-lipid nanocomplex showed minimal transfection in major organs, indicating an excellent safety feature as the gene transfer occurred only at the injection sites, whereas intravenous (i.v.) injection with the same amount of mRNA complexed with a commercial transfection reagent Trans-IT showed luciferase expression in the spleen. In summary, InstantFECT cationic liposomes provide a safe and efficient in vivo locoregional delivery of mRNA and could be a useful tool for basic research and for the development of mRNA-based therapies.