The mixing method used to formulate lipid nanoparticles affects mRNA delivery efficacy and organ tropism.
The mixing method used to formulate lipid nanoparticles affects mRNA delivery efficacy and organ tropism.
复制标题
DOI：
10.1016/j.ejpb.2023.10.006
复制
发表时间：
2023-10
期刊：
影响因子：
--
通讯作者：
Daria M. Strelkova Petersen;Namit Chaudhary;Mariah L. Arral;Ryan M. Weiss;Kathryn A. Whitehead
中科院分区：
文献类型：
--
作者：
Daria M. Strelkova Petersen;Namit Chaudhary;Mariah L. Arral;Ryan M. Weiss;Kathryn A. Whitehead
mRNA是一种多功能药物分子，具有从蛋白质替代疗法到体内基因工程的治疗应用。mRNA的传递通常是通过脂质纳米颗粒完成的，脂质纳米颗粒是通过水和有机溶液的混合而形成的。虽然这在历史上是通过手工混合来完成实验规模的科学，但微流控混合是可扩展的连续制造和批对批控制所必需的。目前，对混合过程如何影响mRNA传递效率的理解有限，特别是在向性方面。为了解决这一知识差距，我们研究了混合类型和微流体混合参数对小鼠脂质纳米颗粒性能的影响。这是通过实验设计方法完成的，使用四种具有不同电离脂质化学性质的纳米颗粒配方。我们发现，每种配方都需要对混合参数进行独特的优化，与手动混合LNPs相比，微流体生成的每个脂质纳米颗粒的总递送效率从100倍到4倍不等。此外，混合参数影响器官向性，与其他器官相比，最有效的配方不成比例地增加肝脏输送。这些数据表明，脂质纳米颗粒生产的混合参数可能需要针对每种独特的化学配方进行优化，从而使转化工作复杂化。此外，必须仔细选择微流体参数，以平衡mRNA的整体递送效率和特定应用的向性要求。
mRNA is a versatile drug molecule with therapeutic applications ranging from protein replacement therapies to in vivo gene engineering. mRNA delivery is often accomplished using lipid nanoparticles, which are formulated via mixing of aqueous and organic solutions. Although this has historically been accomplished by manual mixing for bench scale science, microfluidic mixing is required for scalable continuous manufacturing and batch to batch control. Currently, there is limited understanding on how the mixing process affects mRNA delivery efficacy, particularly in regard to tropism. To address this knowledge gap, we examined the influence of the type of mixing and microfluidic mixing parameters on the performance of lipid nanoparticles in mice. This was accomplished with a Design of Experiment approach using four nanoparticle formulations with varied ionizable lipid chemistry. We found that each formulation required unique optimization of mixing parameters, with the total delivery efficacy of each lipid nanoparticle generated with microfluidics ranging from 100-fold less to 4-fold more than manually mixed LNPs. Further, mixing parameters influenced organ tropism, with the most efficacious formulations disproportionately increasing liver delivery compared to other organs. These data suggest that mixing parameters for lipid nanoparticle production may require optimization for each unique chemical formulation, complicating translational efforts. Further, microfluidic parameters must be chosen carefully to balance overall mRNA delivery efficacy with application-specific tropism requirements.