Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands; Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, the Netherlands; CDL Research, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands.
Center for Translational Immunology, University Medical Center Utrecht, Utrecht University, Utrecht, the Netherlands; Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, the Netherlands.
J Control Release. 2022 Jan;341:475-486. doi: 10.1016/j.jconrel.2021.11.042. Epub 2021 Dec 7.
PEGylation of lipid-based nanoparticles and other nanocarriers is widely used to increase their stability and plasma half-life. However, either pre-existing or de novo formed anti-PEG antibodies can induce hypersensitivity reactions and accelerated blood clearance through binding to the nanoparticle surfaces, leading to activation of the complement system. In this study, we investigated the consequences and mechanisms of complement activation by anti-PEG antibodies interacting with different types of PEGylated lipid-based nanoparticles. By using both liposomes loaded with different (model) drugs and LNPs loaded with mRNA, we demonstrate that complement activation triggered by anti-PEG antibodies can compromise the bilayer/surface integrity, leading to premature drug release or exposure of their mRNA contents to serum proteins. Anti-PEG antibodies also can induce deposition of complement fragments onto the surface of PEGylated lipid-based nanoparticles and induce the release of fluid phase complement activation products. The role of the different complement pathways activated by lipid-based nanoparticles was studied using deficient sera and/or inhibitory antibodies. We identified a major role for the classical complement pathway in the early activation events leading to the activation of C3. Our data also confirm the essential role of amplification of C3 activation by alternative pathway components in the lysis of liposomes. Finally, the levels of pre-existing anti-PEG IgM antibodies in plasma of healthy donors correlated with the degree of complement activation (fixation and lysis) induced upon exposure to PEGylated liposomes and mRNA-LNPs. Taken together, anti-PEG antibodies trigger complement activation by PEGylated lipid-based nanoparticles, which can potentially compromise their integrity, leading to premature drug release or cargo exposure to serum proteins.
聚乙二醇化的脂质纳米粒和其他纳米载体被广泛用于提高其稳定性和血浆半衰期。然而，无论是预先存在的还是新形成的抗聚乙二醇抗体，都可以通过与纳米颗粒表面结合，诱导过敏反应和加速血液清除，从而激活补体系统。在这项研究中，我们研究了抗聚乙二醇抗体与不同类型的聚乙二醇化脂质纳米粒相互作用导致补体激活的后果和机制。通过使用载有不同（模型）药物的脂质体和载有 mRNA 的 LNPs，我们证明了抗聚乙二醇抗体触发的补体激活可以破坏双层/表面完整性，导致药物过早释放或其 mRNA 内容物暴露于血清蛋白中。抗聚乙二醇抗体还可以诱导补体片段沉积在聚乙二醇化脂质纳米粒的表面，并诱导补体激活产物的液相反响释放。使用缺乏血清和/或抑制性抗体研究了脂质纳米粒激活的不同补体途径的作用。我们确定了经典补体途径在导致 C3 激活的早期激活事件中的主要作用。我们的数据还证实了替代途径成分在脂质体裂解中对 C3 激活的放大在替代途径激活中的重要作用。最后，健康供体血浆中预先存在的抗聚乙二醇 IgM 抗体的水平与暴露于聚乙二醇化脂质体和 mRNA-LNPs 后诱导的补体激活（固定和裂解）程度相关。总之，抗聚乙二醇抗体触发聚乙二醇化脂质纳米粒的补体激活，这可能会破坏其完整性，导致药物过早释放或货物暴露于血清蛋白中。
