Notch1和Notch4核心结合结构域蛋白抗体表现出不同的配体结合和抗血管生成特性。

IF 9.2 1区 医学 Q1 PERIPHERAL VASCULAR DISEASE Angiogenesis Pub Date : 2022-11-15 DOI:10.1007/s10456-022-09861-6
Timothy Sargis, Seock-Won Youn, Krishna Thakkar, L. A. Naiche, Na Yoon Paik, Kostandin V. Pajcini, Jan K. Kitajewski
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引用次数: 0

摘要

Notch信号通路是治疗炎症性疾病和癌症的重要治疗靶点。我们之前创建了Notch信号传导的配体特异性抑制剂,由Fc融合到Notch1细胞外结构域的特定EGF样重复序列组成,称为Notch诱饵,它结合配体,阻断Notch信号,并显示出低毒的抗肿瘤活性。然而,对其功能的研究依赖于病毒介导的表达,这阻碍了剂量控制,限制了临床应用。我们已经改进了诱饵设计,以产生基于肽体的Notch抑制剂,其包括Notch1或Notch4的核心结合结构域,EGF样重复10-14。这些Notch蛋白抗体显示出高分泌特性和生产产量,与以前的Notch诱饵相比提高了近100倍。使用表面等离子体共振光谱与共免疫沉淀分析相结合,我们观察到Notch1和Notch4蛋白抗体与Notch配体DLL4和JAG1表现出强烈但不同的结合特性。Notch1和Notch4蛋白抗体均干扰内皮细胞中的Notch信号传导,并在治疗后降低典型Notch靶标的表达。虽然先前的DLL4抑制剂导致超发芽,但在三维体外发芽测定中,Notch1蛋白体减少了血管生成。对新生小鼠施用Notch1蛋白抗体导致视网膜血管系统的径向生长减少,证实了抗血管生成特性。我们得出的结论是,包含EGF样重复10-14的纯化的Notch蛋白抗体与DLL4和JAG1配体结合,并表现出抗血管生成特性。基于其分泌特征、独特的Notch抑制活性和抗血管生成特性,Notch蛋白抗体为治疗性Notch抑制提供了新的机会。
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Notch1 and Notch4 core binding domain peptibodies exhibit distinct ligand-binding and anti-angiogenic properties

The Notch signaling pathway is an important therapeutic target for the treatment of inflammatory diseases and cancer. We previously created ligand-specific inhibitors of Notch signaling comprised of Fc fusions to specific EGF-like repeats of the Notch1 extracellular domain, called Notch decoys, which bound ligands, blocked Notch signaling, and showed anti-tumor activity with low toxicity. However, the study of their function depended on virally mediated expression, which precluded dosage control and limited clinical applicability. We have refined the decoy design to create peptibody-based Notch inhibitors comprising the core binding domains, EGF-like repeats 10–14, of either Notch1 or Notch4. These Notch peptibodies showed high secretion properties and production yields that were improved by nearly 100-fold compared to previous Notch decoys. Using surface plasmon resonance spectroscopy coupled with co-immunoprecipitation assays, we observed that Notch1 and Notch4 peptibodies demonstrate strong but distinct binding properties to Notch ligands DLL4 and JAG1. Both Notch1 and Notch4 peptibodies interfere with Notch signaling in endothelial cells and reduce expression of canonical Notch targets after treatment. While prior DLL4 inhibitors cause hyper-sprouting, the Notch1 peptibody reduced angiogenesis in a 3-dimensional in vitro sprouting assay. Administration of Notch1 peptibodies to neonate mice resulted in reduced radial outgrowth of retinal vasculature, confirming anti-angiogenic properties. We conclude that purified Notch peptibodies comprising EGF-like repeats 10–14 bind to both DLL4 and JAG1 ligands and exhibit anti-angiogenic properties. Based on their secretion profile, unique Notch inhibitory activities, and anti-angiogenic properties, Notch peptibodies present new opportunities for therapeutic Notch inhibition.

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来源期刊
Angiogenesis
Angiogenesis PERIPHERAL VASCULAR DISEASE-
CiteScore
21.90
自引率
8.20%
发文量
37
审稿时长
6-12 weeks
期刊介绍: Angiogenesis, a renowned international journal, seeks to publish high-quality original articles and reviews on the cellular and molecular mechanisms governing angiogenesis in both normal and pathological conditions. By serving as a primary platform for swift communication within the field of angiogenesis research, this multidisciplinary journal showcases pioneering experimental studies utilizing molecular techniques, in vitro methods, animal models, and clinical investigations into angiogenic diseases. Furthermore, Angiogenesis sheds light on cutting-edge therapeutic strategies for promoting or inhibiting angiogenesis, while also highlighting fresh markers and techniques for disease diagnosis and prognosis.
期刊最新文献
Correction: Mitochondrial control of hypoxia-induced pathological retinal angiogenesis. Angiogenesis is limited by LIC1-mediated lysosomal trafficking. Similarities and differences between brain and skin GNAQ p.R183Q driven capillary malformations. Inflammasome activation aggravates choroidal neovascularization. Timed topical dexamethasone eye drops improve mitochondrial function to prevent severe retinopathy of prematurity
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