van Straten Demian, Sork Helena, van de Schepop Luuk, Frunt Rowan, Ezzat Kariem, Schiffelers Raymond M
CDL Research, University Medical Center Utrecht, Utrecht, the Netherlands.
Institute of Technology, University of Tartu, 50411 Tartu, Estonia.
J Control Release. 2024 Sep;373:481-492. doi: 10.1016/j.jconrel.2024.07.044. Epub 2024 Jul 25.
Lipid nanoparticles (LNPs) have successfully entered the clinic for the delivery of mRNA- and siRNA-based therapeutics, most recently as vaccines for COVID-19. Nevertheless, there is a lack of understanding regarding their in vivo behavior, in particular cell targeting. Part of this LNP tropism is based on the adherence of endogenous protein to the particle surface. This protein forms a so-called corona that can change, amongst other things, the circulation time, biodistribution and cellular uptake of these particles. The formation of this protein corona, in turn, is dependent on the nanoparticle properties (e.g., size, charge, surface chemistry and hydrophobicity) as well as the biological environment from which it is derived. With the potential of gene therapy to target virtually any disease, administration sites other than intravenous route are considered, resulting in tissue specific protein coronas. For neurological diseases, intracranial administration of LNPs results in a cerebral spinal fluid derived protein corona, possibly changing the properties of the lipid nanoparticle compared to intravenous administration. Here, the differences between plasma and CSF derived protein coronas on a clinically relevant LNP formulation were studied in vitro. Protein analysis showed that LNPs incubated in human CSF (C-LNPs) developed a protein corona composition that differed from that of LNPs incubated in plasma (P-LNPs). Lipoproteins as a whole, but in particular apolipoprotein E, represented a higher percentage of the total protein corona on C-LNPs than on P-LNPs. This resulted in improved cellular uptake of C-LNPs compared to P-LNPs, regardless of cell origin. Importantly, the higher LNP uptake did not directly translate into more efficient cargo delivery, underlining that further assessment of such mechanisms is necessary. These findings show that biofluid specific protein coronas alter LNP functionality, suggesting that the site of administration could affect LNP efficacy in vivo and needs to be considered during the development of the formulation.
脂质纳米颗粒（LNPs）已成功进入临床，用于递送基于mRNA和siRNA的疗法，最近还用于COVID-19疫苗。然而，人们对其体内行为，尤其是细胞靶向方面仍缺乏了解。这种LNP的部分趋向性是基于内源性蛋白质在颗粒表面的附着。这种蛋白质形成了所谓的冠层，它可以改变这些颗粒的循环时间、生物分布和细胞摄取等。反过来，这种蛋白质冠层的形成取决于纳米颗粒的性质（如大小、电荷、表面化学性质和疏水性）以及其来源的生物环境。由于基因治疗有针对几乎任何疾病的潜力，人们开始考虑静脉途径以外的给药部位，这会导致组织特异性蛋白质冠层的形成。对于神经疾病，颅内注射LNP会产生源自脑脊液的蛋白质冠层，与静脉注射相比，这可能会改变脂质纳米颗粒的性质。在此，我们在体外研究了临床相关LNP制剂上源自血浆和脑脊液的蛋白质冠层之间的差异。蛋白质分析表明，在人脑脊液中孵育的LNP（C-LNPs）形成的蛋白质冠层组成与在血浆中孵育的LNP（P-LNPs）不同。脂蛋白整体上，特别是载脂蛋白E，在C-LNPs的总蛋白质冠层中所占比例高于P-LNPs。这导致C-LNPs与P-LNPs相比，细胞摄取有所改善，无论细胞来源如何。重要的是，较高的LNP摄取并没有直接转化为更有效的货物递送，这突出表明有必要对这些机制进行进一步评估。这些发现表明，生物流体特异性蛋白质冠层会改变LNP的功能，这表明给药部位可能会影响LNP在体内的疗效，在制剂开发过程中需要加以考虑。
