New Anti-Nodal Monoclonal Antibodies Targeting the Nodal Pre-Helix Loop Involved in Cripto-1 Binding.
New Anti-Nodal Monoclonal Antibodies Targeting the Nodal Pre-Helix Loop Involved in Cripto-1 Binding.
复制标题
针对涉及Cripto-1结合的淋巴结前环路的新的抗鼻单克隆抗体。
DOI：
10.3390/ijms160921342
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发表时间：
2015-09-07
影响因子：
5.6
通讯作者：
Sandomenico A
中科院分区：
文献类型：
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作者：
Focà A;Sanguigno L;Focà G;Strizzi L;Iannitti R;Palumbo R;Hendrix MJ;Leonardi A;Ruvo M;Sandomenico A
Nodal是一种属于TGF-β超家族的有效胚胎形态原。通常，它还在共受体Cripto-1存在下结合ALK 4/ActRIIB受体复合物。淋巴结表达在生理上仅限于胚胎组织和人类胚胎干细胞，在正常细胞中不存在，但在几种人类癌症（包括黑色素瘤、乳腺癌和结肠癌）中重新出现。我们的目标是获得能够识别Nodal的主要CBR（Cripto结合区）位点并阻断Cripto-1介导的信号传导的mAb。为了实现这一点，产生了针对hNodal（44-67）的抗体和通过杂交瘤技术产生的mAb。我们已经选择了一种mAb，命名为3D 1，它与全长rhNodal（KD 1.4 nM）强烈相关，并通过Western印迹和FACS分析识别一组人黑色素瘤细胞系中的内源性蛋白。3D 1抑制Nodal-Cripto-1结合并阻断Smad 2/3磷酸化。数据表明，抑制Nodal-Cripto-1轴是针对黑色素瘤的有效治疗方法，并且3D 1是用于阻断Nodal-Cripto介导的肿瘤发展的有前景且令人感兴趣的药剂。这些发现增加了Nodal作为诊断和预后标志物以及作为治疗干预的潜在新靶点的兴趣。
Nodal is a potent embryonic morphogen belonging to the TGF-β superfamily. Typically, it also binds to the ALK4/ActRIIB receptor complex in the presence of the co-receptor Cripto-1. Nodal expression is physiologically restricted to embryonic tissues and human embryonic stem cells, is absent in normal cells but re-emerges in several human cancers, including melanoma, breast, and colon cancer. Our aim was to obtain mAbs able to recognize Nodal on a major CBR (Cripto-Binding-Region) site and to block the Cripto-1-mediated signalling. To achieve this, antibodies were raised against hNodal(44–67) and mAbs generated by the hybridoma technology. We have selected one mAb, named 3D1, which strongly associates with full-length rhNodal (KD 1.4 nM) and recognizes the endogenous protein in a panel of human melanoma cell lines by western blot and FACS analyses. 3D1 inhibits the Nodal-Cripto-1 binding and blocks Smad2/3 phosphorylation. Data suggest that inhibition of the Nodal-Cripto-1 axis is a valid therapeutic approach against melanoma and 3D1 is a promising and interesting agent for blocking Nodal-Cripto mediated tumor development. These findings increase the interest for Nodal as both a diagnostic and prognostic marker and as a potential new target for therapeutic intervention.