Borrull Aurélie, Allard Bertrand, Wijkhuisen Anne, Herbet Amaury, Lamourette Patricia, Birouk Wided, Leiber Denis, Tanfin Zahra, Ducancel Frédéric, Boquet Didier, Couraud Jean-Yves, Robin Philippe
a CEA, iBiTec-S, SPI, Laboratoire d'Ingénierie des Anticorps pour la Santé , Gif-sur-Yvette , France.
b Université Paris Sud-11 , CNRS, UMR 8619, IBBMC , Orsay , France.
MAbs. 2016 Oct;8(7):1371-1385. doi: 10.1080/19420862.2016.1208865. Epub 2016 Jul 8.
Metastatic melanoma is an aggressive cancer with a poor prognostic, and the design of new targeted drugs to treat melanoma is a therapeutic challenge. A promising approach is to produce monoclonal antibodies (mAbs) against the endothelin B receptor (ETB), which is known to be overexpressed in melanoma and to contribute to proliferation, migration and vasculogenic mimicry associated with invasiveness of this cancer. We previously described rendomab-B1, a mAb produced by DNA immunization. It is endowed with remarkable characteristics in term of affinity, specificity and antagonist properties against human ETB expressed by the endothelial cells, but, surprisingly, had poor affinity for ETB expressed by melanoma cells. This characteristic strongly suggested the existence of a tumor-specific ETB form. In the study reported here, we identified a new mAb, rendomab-B4, which, in contrast to rendomab-B1, binds ETB expressed on UACC-257, WM-266-4 and SLM8 melanoma cells. Moreover, after binding to UACC-257 cells, rendomab-B4 is internalized and colocalizes with the endosomal protein EEA-1. Interestingly, rendomab-B4, despite its inability to compete with endothelin binding, is able to inhibit phospholipase C pathway and migration induced by endothelin. By contrast, rendomab-B4 fails to decrease ERK1/2 phosphorylation induced by endothelin, suggesting a biased effect on ETB. These particular properties make rendomab-B4 an interesting tool to analyze ETB-structure/function and a promising starting point for the development of new immunological tools in the field of melanoma therapeutics.
转移性黑色素瘤是一种侵袭性癌症,预后较差,设计新的靶向药物治疗黑色素瘤是一项治疗挑战。一种有前景的方法是制备针对内皮素B受体(ETB)的单克隆抗体(mAb),已知该受体在黑色素瘤中过表达,并与该癌症的侵袭性相关的增殖、迁移和血管生成拟态有关。我们之前描述了通过DNA免疫产生的单克隆抗体rendomab-B1。它在内皮细胞表达的人ETB的亲和力、特异性和拮抗特性方面具有显著特征,但令人惊讶的是,它对黑色素瘤细胞表达的ETB亲和力较差。这一特征强烈表明存在肿瘤特异性的ETB形式。在本文报道的研究中,我们鉴定了一种新的单克隆抗体rendomab-B4,与rendomab-B1不同,它能结合UACC-257、WM-266-4和SLM8黑色素瘤细胞上表达的ETB。此外,与UACC-257细胞结合后,rendomab-B4会内化并与内体蛋白EEA-1共定位。有趣的是,rendomab-B4尽管无法与内皮素结合竞争,但能够抑制内皮素诱导的磷脂酶C途径和迁移。相比之下,rendomab-B4未能降低内皮素诱导的ERK1/2磷酸化,表明对ETB有偏向性作用。这些特殊性质使rendomab-B4成为分析ETB结构/功能的有趣工具,也是黑色素瘤治疗领域开发新免疫工具的有前景的起点。