Żak Magdalena M, Kaur Keerat, Yoo Jimeen, Kurian Ann Anu, Adjmi Matthew, Mainkar Gayatri, Yoon Seonghun, Zangi Lior
Cardiovascular Research Institute, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Pharmaceutics. 2023 Aug 22;15(9):2176. doi: 10.3390/pharmaceutics15092176.
Directly injecting naked or lipid nanoparticle (LNP)-encapsulated modified mRNA (modRNA) allows rapid and efficient protein expression. This non-viral technology has been used successfully in modRNA vaccines against SARS-CoV-2. The main challenges in using modRNA vaccines were the initial requirement for an ultra-cold storage to preserve their integrity and concerns regarding unwanted side effects from this new technology. Here, we showed that naked modRNA maintains its integrity when stored up to 7 days at 4 °C, and LNP-encapsulated modRNA for up to 7 days at room temperature. Naked modRNA is predominantly expressed at the site of injection when delivered into cardiac or skeletal muscle. In comparison, LNP-encapsulated modRNA granted superior protein expression but also additional protein expression beyond the cardiac or skeletal muscle injection site. To overcome this challenge, we developed a skeletal-muscle-specific modRNA translation system (skeletal muscle SMRTs) for LNP-encapsulated modRNA. This system allows controlled protein translation predominantly at the site of injection to prevent potentially detrimental leakage and expression in major organs. Our study revealed the potential of the SMRTs platform for controlled expression of mRNA payload delivered intramuscularly. To conclude, our SMRTs platform for LNP-encapsulated modRNA can provide safe, stable, efficient and targeted gene expression at the site of injection.
直接注射裸露的或脂质纳米颗粒（LNP）包裹的修饰mRNA（modRNA）可实现快速高效的蛋白质表达。这种非病毒技术已成功应用于针对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的modRNA疫苗。使用modRNA疫苗的主要挑战在于最初需要超低温储存以保持其完整性，以及对这项新技术产生的不良副作用的担忧。在此，我们表明，裸露的modRNA在4℃下储存长达7天仍能保持其完整性，而LNP包裹的modRNA在室温下可储存长达7天。将裸露的modRNA注入心脏或骨骼肌时，主要在注射部位表达。相比之下，LNP包裹的modRNA不仅能实现更高水平的蛋白质表达，还能在心脏或骨骼肌注射部位以外产生额外的蛋白质表达。为克服这一挑战，我们为LNP包裹的modRNA开发了一种骨骼肌特异性modRNA翻译系统（骨骼肌SMRTs）。该系统可使蛋白质翻译主要在注射部位受到控制，以防止在主要器官中出现潜在有害的泄漏和表达。我们的研究揭示了SMRTs平台在肌肉注射时控制mRNA载荷表达的潜力。总之，我们用于LNP包裹的modRNA的SMRTs平台能够在注射部位提供安全、稳定、高效且靶向性的基因表达。
