Gelatin Nanoparticles for Complexation and Enhanced Cellular Delivery of mRNA.
Gelatin Nanoparticles for Complexation and Enhanced Cellular Delivery of mRNA.
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DOI：
10.3390/nano12193423
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发表时间：
2022-09-29
期刊：
影响因子：
--
通讯作者：
Leeuwenburgh SCG
中科院分区：
文献类型：
--
作者：
Andrée L;Oude Egberink R;Dodemont J;Hassani Besheli N;Yang F;Brock R;Leeuwenburgh SCG
信使RNA（mRNA）作为疫苗接种和蛋白质替代疗法的一种方式越来越受到人们的关注。在再生医学中，mRNA介导的生长因子表达已显示出有希望的结果。与蛋白质递送相反，成功的mRNA递送需要载体来诱导细胞摄取和随后的内体逃逸以到达其最终目的地，核糖体。目前的非病毒载体如基于脂质或聚合物的纳米颗粒已成功用于表达mRNA编码的蛋白质。然而，为了促进mRNA在再生医学中的应用，需要评估mRNA与通常应用于该领域的生物材料的相容性。在此，我们研究了与明胶纳米颗粒（GNP）复合的mRNA的复合、细胞摄取和完整性的维持。为此，合成具有正、中性或负表面电荷的GNP以评估它们结合mRNA并将其转运到细胞中的能力。正电荷的GNP表现出最高的结合亲和力，并运输大量的mRNA进入前成骨细胞，通过共聚焦显微镜使用荧光标记的mRNA进行评估。此外，GNP结合的mRNA保持稳定。然而，没有检测到mRNA编码的蛋白质的表达，这可能与GNP的内体逃逸和/或mRNA释放不足有关。我们的研究结果表明，明胶基纳米材料与mRNA以电荷依赖性方式相互作用，并介导细胞摄取。这些结果为引入进一步的功能性以产生内体释放奠定了基础。
Messenger RNA (mRNA) is increasingly gaining interest as a modality in vaccination and protein replacement therapy. In regenerative medicine, the mRNA-mediated expression of growth factors has shown promising results. In contrast to protein delivery, successful mRNA delivery requires a vector to induce cellular uptake and subsequent endosomal escape to reach its end destination, the ribosome. Current non-viral vectors such as lipid- or polymer-based nanoparticles have been successfully used to express mRNA-encoded proteins. However, to advance the use of mRNA in regenerative medicine, it is required to assess the compatibility of mRNA with biomaterials that are typically applied in this field. Herein, we investigated the complexation, cellular uptake and maintenance of the integrity of mRNA complexed with gelatin nanoparticles (GNPs). To this end, GNPs with positive, neutral or negative surface charge were synthesized to assess their ability to bind and transport mRNA into cells. Positively charged GNPs exhibited the highest binding affinity and transported substantial amounts of mRNA into pre-osteoblastic cells, as assessed by confocal microscopy using fluorescently labeled mRNA. Furthermore, the GNP-bound mRNA remained stable. However, no expression of mRNA-encoded protein was detected, which is likely related to insufficient endosomal escape and/or mRNA release from the GNPs. Our results indicate that gelatin-based nanomaterials interact with mRNA in a charge-dependent manner and also mediate cellular uptake. These results create the basis for the incorporation of further functionality to yield endosomal release.
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