Intracellular delivery of NF-κB inhibitor peptide utilizing small extracellular vesicles for the application of anti-inflammatory therapy.
Intracellular delivery of NF-κB inhibitor peptide utilizing small extracellular vesicles for the application of anti-inflammatory therapy.
复制标题
利用细胞外囊泡在细胞内递送 NF-κB 抑制肽，用于抗炎治疗。
DOI：
10.1016/j.jconrel.2020.09.001
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Takakura Y.
中科院分区：
文献类型：
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作者：
Takenaka M;Takahashi Y;Takakura Y.
巨噬细胞中NF-κB的过度活化有助于炎症性疾病的发生和加重。NEMO结合结构域（NBD）肽是一种NF-κB抑制剂肽，其在胞质溶胶中与NEMO（IκB激酶（IKK）复合物的组分之一）结合并抑制IKK激酶活性。由于该性质，预期NBD肽抑制巨噬细胞中的NF-κB活化。在这项研究中，我们开发了一种基于小细胞外囊泡（sEV）的NBD递送载体，sEV是从细胞释放的膜囊泡。我们构建了包含Gag（sEV嗜性蛋白）和一个、三个或六个重复的NBD肽（分别为Gag-1 NBD、Gag-3 NBD和Gag-6 NBD）的融合蛋白，以将NBD肽装载到sEV的内部空间，并尝试使用Gag-NBD装载的sEV（nNBD-sEV）将NBD肽细胞内递送到巨噬细胞。nNBD-sEV以重复数依赖性方式显著抑制LPS诱导的巨噬细胞NF-κB通路相关蛋白的磷酸化。此外，它们对NF-κ B依赖的促炎介质如TNFα、CXCL 10、iNOS和NO的表达产生抑制作用。总的来说，我们的结果表明，含有NBD的sEV可用于治疗炎性疾病，因为它们能够有效地将肽递送到巨噬细胞胞质溶胶。
Excessive activation of NF-κB in macrophages contributes to the onset and exacerbation of inflammatory disorders. The NEMO binding domain (NBD) peptide is an NF-κB inhibitor peptide that binds to NEMO, one of components of the IκB kinase (IKK) complex, and inhibits the IKK kinase activity, in the cytosol. Because of this property, the NBD peptide is expected to inhibit NF-κB activation in macrophages. In this study, we developed a delivery carrier for NBD based on small extracellular vesicles (sEVs), which are membrane vesicles released from cells. We constructed fusion proteins comprising Gag (an sEV tropic protein) and one, three, or six repeats of NBD peptide (Gag-1NBD, Gag-3NBD, and Gag-6NBD, respectively) to load the NBD peptide to the inner space of the sEVs, and attempted the intracellular delivery of the NBD peptide to macrophages using Gag-NBD-loaded sEVs (nNBD-sEVs). The nNBD-sEVs significantly inhibited LPS-induced phosphorylation of NF-κB pathway-related proteins in macrophages in a repeat number-dependent manner. Moreover, they exerted inhibitory effects on the NF-κB-dependent expression of proinflammatory mediators such as TNFα, CXCL10, iNOS, and NO. Collectively, our results indicate that NBD-containing sEVs can be used for the treatment of inflammatory diseases owing to their ability to effectively deliver peptides to the macrophage cytosol.