Cell penetrating peptides coupled to an endothelial nitric oxide synthase sequence alter endothelial permeability.
Cell penetrating peptides coupled to an endothelial nitric oxide synthase sequence alter endothelial permeability.
复制标题
与内皮一氧化氮合酶序列偶联的细胞穿透肽可改变内皮通透性。
DOI：
10.1080/21688370.2021.2017226
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发表时间：
2022
期刊：
影响因子：
3.1
通讯作者：
Stark,RyanJ
中科院分区：
文献类型：
--
作者：
Koch,StephenR;Stark,RyanJ
通过使用细胞穿透肽（CPP）序列将货物递送至细胞是靶向治疗的丰富研究领域。具体到内皮，覆盖体内每个血管的细胞层，一种关键酶内皮一氧化氮合酶（eNOS）的丢失或改变，已知在严重的感染性挑战期间有助于内皮健康。虽然eNOS的有益作用通常被认为是通过产生一氧化氮介导的，但理论上一些保护作用是通过eNOS经由五碱基RRKRK基序结合到调节途径。我们假设使用普通CPP递送eNOS-RRKRK肽序列将允许针对革兰氏阴性脂多糖（LPS）的保护。eNOS-RRKRK序列与CPP触角足（AP）的组合降低了培养的人微血管内皮细胞（HMVEC）中LPS诱导的渗透性的影响，如通过跨内皮电阻（TEER）所测量的。细胞因子的产生也有适度的减少，但是观察到单独的AP显著损害LPS诱导的内皮通透性和细胞因子的产生。相比之下，CPP转录反式激活因子（达特）本身并不显著改变内皮炎症。当达特与eNOS-RRKRK序列偶联时，仍然证明了对LPS诱导的渗透性的保护，但是细胞因子的产生没有减少。这些数据表明，eNOS的RRKRK序列可以提供一些NO非依赖性的保护，以对抗LPS介导的内皮炎症，然而，保护的程度高度依赖于用于货物递送的CPP的类型。
Delivery of cargo to cells through the use of cell-penetrating peptide (CPP) sequences is an area of rich investigation for targeted therapeutics. Specific to the endothelium, the layer of cells that cover every blood vessel in the body, the loss or alteration of a key enzyme, endothelial nitric oxide synthase (eNOS), is known to contribute to endothelial health during severe, infectious challenge. While the beneficial effects of eNOS are often thought to be mediated through the generation of nitric oxide, some protection is theorized to be through eNOS binding to regulatory pathways via a pentabasic RRKRK motif. We hypothesized that delivery of the eNOS-RRKRK peptide sequence using common CPPs would allow protection against gram-negative lipopolysaccharide (LPS). Combination of the eNOS-RRKRK sequence to the CPP antennapedia (AP) reduced the impact of LPS-induced permeability in cultured human microvascular endothelial cells (HMVECs) as measured by transendothelial electrical resistance (TEER). There was also a modest reduction in cytokine production, however it was observed that AP alone significantly impaired LPS-induced endothelial permeability and cytokine production. In comparison, the CPP trans-activator of transcription (TAT) did not significantly alter endothelial inflammation by itself. When TAT was coupled to the eNOS-RRKRK sequence, protection against LPS-induced permeability was still demonstrated, however cytokine production was not reduced. These data demonstrate that the RRKRK sequence of eNOS can offer some NO-independent protection against LPS-mediated endothelial inflammation, however the degree of protection is highly dependent on the type of CPP utilized for cargo delivery.