A human antibody against human endothelin receptor type A that exhibits antitumor potency

Man Seok Ju; Hye Mi Ahn; Seong Gu Han; Sanghwan Ko; Jung Hyun Na; Migyeong Jo; Chung Su Lim; Byoung Joon Ko; Yeon Gyu Yu; Won Kyu Lee; Youn Jae Kim; Sang Taek Jung

doi:10.1038/s12276-021-00678-9

A human antibody against human endothelin receptor type A that exhibits antitumor potency

Man Seok Ju, Hye Mi Ahn, Seong Gu Han, Sanghwan Ko, Jung Hyun Na, Migyeong Jo, Chung Su Lim, Byoung Joon Ko, Yeon Gyu Yu, Won Kyu Lee, Youn Jae Kim, Sang Taek Jung

Department of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

15 Citations (Scopus)

Abstract

Endothelin receptor A (ET_A), 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 ET_A 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-ET_A antibody (AG8) exhibiting high specificity for ET_A in the β-arrestin Tango assay and effective inhibitory activity against the ET-1-induced signaling cascade via ET_A using either a CHO-K1 cell line stably expressing human ET_A 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 ET_A-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-ET_A antibody isolated in this study could be used as a potential therapeutic for cancers, including colorectal cancer.

Original language	English
Pages (from-to)	1437-1448
Number of pages	12
Journal	Experimental and Molecular Medicine
Volume	53
Issue number	9
DOIs	https://doi.org/10.1038/s12276-021-00678-9
Publication status	Published - 2021 Sept

Bibliographical note

Funding Information:
This work was supported by grants from the Bio & Medical Technology Development Programs (2017M3A9C8060541, 2017M3A9C8060558, 2017M3A9C8060560, and 2020M3E5E2037775) and the Basic Science Research Programs (2019R1A4A1029000, 2019R1A6A3A01097279, 2018R1D1A1B07051154, and 2019R1A2C1086258) through the National Research Foundation of Korea funded by the Ministry of Science and ICT and by a National Cancer Center grant (NCC-2010250).

Publisher Copyright:
© 2021, The Author(s).

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Clinical Biochemistry

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/s12276-021-00678-9

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@article{d377ba26f10b4d908d5eae7df887c596,

title = "A human antibody against human endothelin receptor type A that exhibits antitumor potency",

abstract = "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.",

author = "Ju, {Man Seok} and Ahn, {Hye Mi} and Han, {Seong Gu} and Sanghwan Ko and Na, {Jung Hyun} and Migyeong Jo and Lim, {Chung Su} and Ko, {Byoung Joon} and Yu, {Yeon Gyu} and Lee, {Won Kyu} and Kim, {Youn Jae} and Jung, {Sang Taek}",

note = "Funding Information: This work was supported by grants from the Bio & Medical Technology Development Programs (2017M3A9C8060541, 2017M3A9C8060558, 2017M3A9C8060560, and 2020M3E5E2037775) and the Basic Science Research Programs (2019R1A4A1029000, 2019R1A6A3A01097279, 2018R1D1A1B07051154, and 2019R1A2C1086258) through the National Research Foundation of Korea funded by the Ministry of Science and ICT and by a National Cancer Center grant (NCC-2010250). Publisher Copyright: {\textcopyright} 2021, The Author(s).",

year = "2021",

month = sep,

doi = "10.1038/s12276-021-00678-9",

language = "English",

volume = "53",

pages = "1437--1448",

journal = "Experimental and Molecular Medicine",

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T1 - A human antibody against human endothelin receptor type A that exhibits antitumor potency

AU - Ju, Man Seok

AU - Ahn, Hye Mi

AU - Han, Seong Gu

AU - Ko, Sanghwan

AU - Na, Jung Hyun

AU - Jo, Migyeong

AU - Lim, Chung Su

AU - Ko, Byoung Joon

AU - Yu, Yeon Gyu

AU - Lee, Won Kyu

AU - Kim, Youn Jae

AU - Jung, Sang Taek

N1 - Funding Information: This work was supported by grants from the Bio & Medical Technology Development Programs (2017M3A9C8060541, 2017M3A9C8060558, 2017M3A9C8060560, and 2020M3E5E2037775) and the Basic Science Research Programs (2019R1A4A1029000, 2019R1A6A3A01097279, 2018R1D1A1B07051154, and 2019R1A2C1086258) through the National Research Foundation of Korea funded by the Ministry of Science and ICT and by a National Cancer Center grant (NCC-2010250). Publisher Copyright: © 2021, The Author(s).

PY - 2021/9

Y1 - 2021/9

N2 - 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.

AB - 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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U2 - 10.1038/s12276-021-00678-9

DO - 10.1038/s12276-021-00678-9

M3 - Article

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AN - SCOPUS:85115989787

SN - 1226-3613

VL - 53

SP - 1437

EP - 1448

JO - Experimental and Molecular Medicine

JF - Experimental and Molecular Medicine

IS - 9

ER -

A human antibody against human endothelin receptor type A that exhibits antitumor potency

Abstract

Bibliographical note

ASJC Scopus subject areas

UN SDGs

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