Identification and characterization of a mirror-image oligonucleotide that binds and neutralizes sphingosine 1-phosphate, a central mediator of angiogenesis.
Identification and characterization of a mirror-image oligonucleotide that binds and neutralizes sphingosine 1-phosphate, a central mediator of angiogenesis.
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结合和中和1-磷酸鞘氨酸的镜像寡核苷酸的识别和表征，这是血管生成的中心介体。
DOI：
10.1042/bj20131422
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发表时间：
2014-08-15
期刊：
影响因子：
--
通讯作者：
Klussmann S
中科院分区：
文献类型：
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作者：
Purschke WG;Hoehlig K;Buchner K;Zboralski D;Schwoebel F;Vater A;Klussmann S
鞘脂S1P（鞘磷脂1-磷酸）参与了许多病理生理状况，如癌症、自身免疫性疾病和纤维化。它通过一组5个g蛋白偶联受体在细胞外起作用，但它在细胞内的作用也有很好的记录。采用体外筛选技术，我们鉴定出一种l -适配体（Spiegelmer®），用于S1P，指定为NOX-S93。NOX-S93与S1P的结合亲和力Kd值为4.3 nM。Spiegelmer®与二氢s1p有相同的结合，但与相关脂质鞘氨醇、溶血磷脂酸、神经酰胺、神经酰胺-1-磷酸或鞘氨醇磷酸胆碱无交叉反应性。在稳定转染表达S1P受体S1PR1或S1PR3的CHO（中国仓鼠卵巢）细胞系中，NOX-S93分别抑制S1P介导的β-阻滞蛋白募集和细胞内钙释放，IC50值在低纳摩尔范围内。在使用原代人内皮细胞进行的细胞血管生成实验中，NOX-S93有效地阻断了S1P以及生长因子VEGF-A（血管内皮生长因子-a）、FGF-2（成纤维细胞生长因子-2）和IGF-1（胰岛素样生长因子-1）的促血管生成活性。这些数据为细胞外S1P作为血管生成的中心介质的相关性提供了进一步的证据，表明药物S1P中和是目前抗血管生成方法的一种有希望的治疗选择。镜像适配体NOX-S93，通过体外选择鉴定为特异性结合S1P，抑制S1P受体的信号传导。NOX-S93抑制S1P和其他生长因子的促血管生成活性，表明S1P中和是一种很有前途的抗血管生成方法。
The sphingolipid S1P (sphingosine 1-phosphate) is known to be involved in a number of pathophysiological conditions such as cancer, autoimmune diseases and fibrosis. It acts extracellularly through a set of five G-protein-coupled receptors, but its intracellular actions are also well documented. Employing in vitro selection techniques, we identified an L-aptamer (Spiegelmer®) to S1P designated NOX-S93. The binding affinity of NOX-S93 to S1P had a Kd value of 4.3 nM. The Spiegelmer® shows equal binding to dihydro-S1P, but no cross-reactivity to the related lipids sphingosine, lysophosphatidic acid, ceramide, ceramide-1-phosphate or sphingosine phosphocholine. In stably transfected CHO (Chinese-hamster ovary) cell lines expressing the S1P receptors S1PR1 or S1PR3, NOX-S93 inhibits S1P-mediated β-arrestin recruitment and intracellular calcium release respectively, with IC50 values in the low nanomolar range. The pro-angiogenic activity of S1P, and of the growth factors VEGF-A (vascular endothelial growth factor-A), FGF-2 (fibroblast growth factor-2) and IGF-1 (insulin-like growth factor-1), was effectively blocked by NOX-S93 in a cellular angiogenesis assay employing primary human endothelial cells. These data provide further evidence for the relevance of extracellular S1P as a central mediator of angiogenesis, suggesting pharmacological S1P neutralization as a promising treatment alternative to current anti-angiogenesis approaches. The mirror-image aptamer NOX-S93, identified by in vitro selection to bind specifically to S1P, inhibits signalling at S1P receptors. NOX-S93 inhibits the pro-angiogenic activity of S1P and other growth factors, revealing S1P neutralization as a promising anti-angiogenesis approach.