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KMID : 0620920210530091437
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Experimental & Molecular Medicine
2021 Volume.53 No. 9 p.1437 ~ p.1448
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A human antibody against human endothelin receptor type A that exhibits antitumor potency
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Ju Man-Seok
Ahn Hye-Mi
Han Seong-Gu
Ko Sang-Hwan
Na Jung-Hyun
Jo Mi-Gyeong
Lim Chung-Su
Ko Byoung-Joon
Yu Yeon-Gyu
Lee Won-Kyu
Kim Youn-Jae
Jung Sang-Taek
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Abstract
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Endothelin receptor A (ETA), a class A G-protein-coupled receptor (GPCR), is involved in the progression and metastasis of colorectal, breast, lung, ovarian, and prostate cancer. We overexpressed and purified human endothelin receptor type A in Escherichia coli and reconstituted it with lipid and membrane scaffold proteins to prepare an ETA nanodisc as a functional antigen with a structure similar to that of native GPCR. By screening a human naive immune single-chain variable fragment phage library constructed in-house, we successfully isolated a human anti-ETA antibody (AG8) exhibiting high specificity for ETA in the β-arrestin Tango assay and effective inhibitory activity against the ET-1-induced signaling cascade via ETA using either a CHO-K1 cell line stably expressing human ETA or HT-29 colorectal cancer cells, in which AG8 exhibited IC50 values of 56 and 51?nM, respectively. In addition, AG8 treatment repressed the transcription of inhibin βA and reduced the ETA-induced phosphorylation of protein kinase B and extracellular regulated kinase. Furthermore, tumor growth was effectively inhibited by AG8 in a colorectal cancer mouse xenograft model. The human anti-ETA antibody isolated in this study could be used as a potential therapeutic for cancers, including colorectal cancer.
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KEYWORD
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Antibody therapy, High-throughput screening
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