Employing Macrophage-Derived Microvesicle for Kidney-Targeted Delivery of Dexamethasone: An Efficient Therapeutic Strategy against Renal Inflammation and Fibrosis
Employing Macrophage-Derived Microvesicle for Kidney-Targeted Delivery of Dexamethasone: An Efficient Therapeutic Strategy against Renal Inflammation and Fibrosis
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DOI：
10.7150/thno.33520
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发表时间：
2019-01-01
期刊：
影响因子：
12.4
通讯作者：
Liu, Bi-Cheng
中科院分区：
文献类型：
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作者：
Tang, Tao-Tao;Lv, Lin-Li;Liu, Bi-Cheng
尽管几十年来糖皮质激素一直是治疗肾脏疾病的主要药物，但其剂量依赖性副作用在很大程度上限制了其临床使用。微泡 (MV) 是几乎所有细胞都会产生的基于脂质的膜结合小颗粒。在这里，我们证明RAW 264.7巨噬细胞来源的MV可以作为载体，有效地递送地塞米松（命名为MV-DEX），靶向发炎的肾脏。方法：RAW巨噬细胞与地塞米松一起孵育，然后通过离心方法从上清液中分离MV-DEX。使用纳米粒子追踪分析、透射电子显微镜、蛋白质印迹和高效液相色谱来分析MV-DEX的特性。应用 LC-MS/MS 研究 MV-DEX 的蛋白质组成。基于LPS或阿霉素（ADR）诱导的肾病小鼠模型或肾小球内皮细胞的体外培养，检查了MV-DEX的炎症靶向特性和治疗效果。最后，我们评估了长期糖皮质激素治疗对 MV-DEX 治疗小鼠的副作用。结果：蛋白质组学分析揭示了 MV-DEX 表面不同的整合素表达模式，其中整合素 α(L)β(2) (LFA-1) 和 α(4)beta(1) (VAL-4) 使它们能够粘附在发炎的肾脏上。与游离 DEX 治疗相比，等摩尔剂量的 MV-DEX 显着减轻了 LPS 或 ADR 诱导肾病小鼠模型中的肾损伤，并增强了对肾脏炎症和纤维化的治疗效果。在体外，含有约五分之一剂量的游离DEX的MV-DEX通过抑制NF-kappa B活性实现了显着的抗炎功效。从机制上讲，MV-DEX可以包装糖皮质激素受体并将其递送至肾细胞，从而增加受体的细胞水平并提高细胞对糖皮质激素的敏感性。值得注意的是，在 MV 中递送 DEX 显着降低了慢性糖皮质激素治疗的副作用（例如高血糖、HPA 轴抑制）。结论：总而言之，巨噬细胞衍生的 MV 有效地将 DEX 递送到发炎的肾脏中，并表现出卓越的抑制肾脏炎症和纤维化的能力，而没有明显的糖皮质激素副作用。我们的研究结果证明了一种新型药物输送策略的有效性和安全性，具有广阔的临床应用前景。
Although glucocorticoids are the mainstays in the treatment of renal diseases for decades, the dose dependent side effects have largely restricted their clinical use. Microvesicles (MVs) are small lipid-based membrane-bound particles generated by virtually all cells. Here we show that RAW 264.7 macrophage cell-derived MVs can be used as vectors to deliver dexamethasone (named as MV-DEX) targeting the inflamed kidney efficiently.Methods: RAW macrophages were incubated with dexamethasone and then MV-DEX was isolated from the supernatants by centrifugation method. Nanoparticle tracking analysis, transmission electron microscopy, western blot and high-performance liquid chromatography were used to analyze the properties of MV-DEX. The LC-MS/MS was applied to investigate the protein compositions of MV-DEX. Based on the murine models of LPS-or Adriamycin (ADR)-induced nephropathy or in-vitro culture of glomerular endothelial cells, the inflammation-targeting characteristics and the therapeutic efficacy of MV-DEX was examined. Finally, we assessed the side effects of chronic glucocorticoid therapy in MV-DEX-treated mice.Results: Proteomic analysis revealed distinct integrin expression patterns on the MV-DEX surface, in which the integrin alpha(L)beta(2) (LFA-1) and alpha(4)beta(1) (VAL-4) enabled them to adhere to the inflamed kidney. Compared to free DEX treatment, equimolar doses of MV-DEX significantly attenuated renal injury with an enhanced therapeutic efficacy against renal inflammation and fibrosis in murine models of LPS-or ADR-induced nephropathy. In vitro, MV-DEX with about one-fifth of the doses of free DEX achieved significant anti-inflammatory efficacy by inhibiting NF-kappa B activity. Mechanistically, MV-DEX could package and deliver glucocorticoid receptors to renal cells, thereby, increasing cellular levels of the receptor and improving cell sensitivity to glucocorticoids. Notably, delivering DEX in MVs significantly reduced the side effects of chronic glucocorticoid therapy (e.g., hyperglycemia, suppression of HPA axis).Conclusion: In summary, macrophage-derived MVs efficiently deliver DEX into the inflamed kidney and exhibit a superior capacity to suppress renal inflammation and fibrosis without apparent glucocorticoid adverse effects. Our findings demonstrate the effectiveness and security of a novel drug delivery strategy with promising clinical applications.