Technical University of Denmark , Department of Health Technology , 2800 Kongens Lyngby , Denmark.
Mabtech AB , 131 52 Nacka Strand , Sweden.
Bioconjug Chem. 2019 Oct 16;30(10):2634-2646. doi: 10.1021/acs.bioconjchem.9b00552. Epub 2019 Sep 19.
The application of reconstituted high-density lipoproteins (rHDL) as a drug-carrier has during the past decade been established as a promising approach for effective receptor-mediated drug delivery, and its ability to target tumors has recently been confirmed in a clinical trial. The rHDL mimics the endogenous HDL, which is known to be highly dynamic and undergo extensive enzyme-mediated remodulations. Hence, to reveal the physiological rHDL stability, a thorough characterization of the dynamics of rHDL in biologically relevant environments is needed. We employ a size-exclusion chromatography (SEC) method to evaluate the dynamics of discoidal rHDL in fetal bovine serum (FBS), where we track both the rHDL lipids (by the fluorescence from lipid-conjugated fluorophores) and apoA-I (by human apoA-I ELISA). We show by using lipoprotein depleted FBS and isolated lipoproteins that rHDL lipids can be transferred to endogenous lipoproteins via direct interactions in a nonenzymatic process, resulting in rHDL compositional- and size-remodeling. This type of dynamics could lead to misinterpretations of fluorescence-based rHDL uptake studies due to desorption of labile lipophilic fluorophores or off-target side effects due to desorption of incorporated drugs. Importantly, we show how the degree of rHDL remodeling can be controlled by the compositional design of the rHDL. Understanding the correlation between the molecular properties of the rHDL constituents and their collective dynamics is essential for improving the rHDL-based drug delivery platform. Taken together, our work highlights the need to carefully consider the compositional design of rHDL and test its stability in a biological relevant environment, when developing rHDL for drug delivery purposes.
在过去的十年中，将重组高密度脂蛋白（rHDL）作为药物载体的应用已被确立为一种有前途的有效受体介导药物递送方法，最近在一项临床试验中证实了其靶向肿瘤的能力。rHDL 模拟内源性 HDL，已知其具有高度动态性，并经历广泛的酶介导重塑。因此，为了揭示生理 rHDL 的稳定性，需要对生物相关环境中 rHDL 的动力学进行彻底的表征。我们采用尺寸排阻色谱（SEC）方法评估圆盘状 rHDL 在胎牛血清（FBS）中的动力学，在此过程中，我们跟踪 rHDL 脂质（通过脂质缀合荧光团的荧光）和载脂蛋白 A-I（通过人载脂蛋白 A-I ELISA）。我们通过使用脂蛋白耗尽的 FBS 和分离的脂蛋白表明，rHDL 脂质可以通过非酶过程中的直接相互作用转移到内源性脂蛋白中，导致 rHDL 组成和大小重塑。这种动力学可能导致基于荧光的 rHDL 摄取研究的误解，因为亲脂性荧光团的解吸或由于掺入药物的解吸而导致脱靶副作用。重要的是，我们展示了如何通过 rHDL 的组成设计来控制 rHDL 重塑的程度。了解 rHDL 成分的分子特性与其集体动力学之间的相关性对于改进基于 rHDL 的药物递送平台至关重要。总之，我们的工作强调了在开发用于药物递送的 rHDL 时，需要仔细考虑 rHDL 的组成设计并在生物相关环境中测试其稳定性。
