PDF(Pubmed)
Abstract：
mRNA applications have undergone unprecedented applications-from vaccination to cell therapy. Natural killer (NK) cells are recognized to have a significant potential in immunotherapy. NK-based cell therapy has drawn attention as allogenic graft with a minimal graft-versus-host risk leading to easier off-the-shelf production. NK cells can be engineered with either viral vectors or electroporation, involving high costs, risks, and toxicity, emphasizing the need for alternative way as mRNA technology. We successfully developed, screened, and optimized novel lipid-based platforms based on imidazole lipids. Formulations are produced by microfluidic mixing and exhibit a size of approximately 100 nm with a polydispersity index of less than 0.2. They are able to transfect NK-92 cells, KHYG-1 cells, and primary NK cells with high efficiency without cytotoxicity, while Lipofectamine Messenger Max and D-Lin-MC3 lipid nanoparticle-based formulations do not. Moreover, the translation of non-modified mRNA was higher and more stable in time compared with a modified one. Remarkably, the delivery of therapeutically relevant interleukin 2 mRNA resulted in extended viability together with preserved activation markers and cytotoxic ability of both NK cell lines and primary NK cells. Altogether, our platforms feature all prerequisites needed for the successful deployment of NK-based therapeutic strategies.
摘要：
mRNA的应用经历了前所未有的应用-从疫苗接种到细胞治疗。自然杀伤(NK)细胞被认为在免疫疗法中具有显著的潜力。基于NK的细胞疗法已引起人们的注意，因为同种异体移植物具有最小的移植物抗宿主风险，从而导致更容易的现成生产。NK细胞可以用病毒载体或电穿孔进行工程改造，涉及高成本，风险,和毒性，强调需要替代方式作为mRNA技术。我们成功开发,筛选,并优化了基于咪唑脂质的新型脂质平台。制剂通过微流体混合制备,并表现出约100nm的尺寸,多分散指数小于0.2。他们能够转染NK-92细胞，KHYG-1细胞,和高效无细胞毒性的原代NK细胞，而LipofectamineMessengerMax和D-Lin-MC3基于脂质纳米颗粒的制剂则没有。此外，与修饰的mRNA相比，未修饰的mRNA的翻译更高，时间更稳定。值得注意的是,治疗相关的白介素2mRNA的递送导致NK细胞系和原代NK细胞的生存力以及保留的活化标记和细胞毒性。总之,我们的平台具备成功部署基于NK的治疗策略所需的所有先决条件.
