Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1

Abstract The development of effective immunotherapies for solid tumors remains a significant challenge. In previous studies, we identified PVT1, a long non-coding RNA, with the peptide HF10 derived from PVT1, presented by HLA-A24. This study aims to develop a single-chain variable fragment (scFv) th...

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Main Authors: Hirotaka Kato, Tomohide Tsukahara, Kenji Murata, Hiromu Nishikata, Yuka Mizue, Takashi Sasaya, Terufumi Kubo, Takayuki Kanaseki, Yoshihiko Hirohashi, Atsushi Oyagi, Tatsuo Maeda, Akihiro Miyazaki, Toshihiko Torigoe
Format: Article
Language:English
Published: Springer 2025-03-01
Series:Cancer Immunology, Immunotherapy
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Online Access:https://doi.org/10.1007/s00262-025-03976-7
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author Hirotaka Kato
Tomohide Tsukahara
Kenji Murata
Hiromu Nishikata
Yuka Mizue
Takashi Sasaya
Terufumi Kubo
Takayuki Kanaseki
Yoshihiko Hirohashi
Atsushi Oyagi
Tatsuo Maeda
Akihiro Miyazaki
Toshihiko Torigoe
author_facet Hirotaka Kato
Tomohide Tsukahara
Kenji Murata
Hiromu Nishikata
Yuka Mizue
Takashi Sasaya
Terufumi Kubo
Takayuki Kanaseki
Yoshihiko Hirohashi
Atsushi Oyagi
Tatsuo Maeda
Akihiro Miyazaki
Toshihiko Torigoe
author_sort Hirotaka Kato
collection DOAJ
description Abstract The development of effective immunotherapies for solid tumors remains a significant challenge. In previous studies, we identified PVT1, a long non-coding RNA, with the peptide HF10 derived from PVT1, presented by HLA-A24. This study aims to develop a single-chain variable fragment (scFv) that specifically recognizes the HLA-A24/HF10 complex (HF10 scFv) and to evaluate its specificity, reactivity, and therapeutic potential as part of a T cell engaging bispecific antibody (HF10xCD3) in vitro and in vivo. Using a scFv phage display library, we screened for scFv clones targeting the HLA-A24/HF10 peptide complex. The selected HF10 scFv was engineered into an IgG1 format (HF10-hIgG1), which demonstrated high affinity (KD = 2.18 × 10⁻⁸ M) and specific detection of the HLA-A24/HF10 complex on HLA-A24( +)/PVT1( +) tumor cell lines. Furthermore, HF10 scFv was incorporated into a T cell engaging bispecific antibody (HF10xCD3), which induced cytotoxicity in these tumor cell lines. In a mouse xenograft model, HF10xCD3 administration exhibited significant anti-tumor activity. In conclusion, HF10xCD3 represents a promising candidate for immunotherapy targeting solid tumors.
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spelling doaj-art-65edf236c6d3473186febd86db5640e52025-08-20T02:49:23ZengSpringerCancer Immunology, Immunotherapy1432-08512025-03-0174411110.1007/s00262-025-03976-7Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1Hirotaka Kato0Tomohide Tsukahara1Kenji Murata2Hiromu Nishikata3Yuka Mizue4Takashi Sasaya5Terufumi Kubo6Takayuki Kanaseki7Yoshihiko Hirohashi8Atsushi Oyagi9Tatsuo Maeda10Akihiro Miyazaki11Toshihiko Torigoe12Department of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityOncology Clinical Exploratory Research II, Ono Pharmaceutical, Co., Ltd.Research Center of Oncology Group VI, Ono Pharmaceutical, Co., Ltd.Department of Pathology, School of Medicine, Sapporo Medical UniversityDepartment of Pathology, School of Medicine, Sapporo Medical UniversityAbstract The development of effective immunotherapies for solid tumors remains a significant challenge. In previous studies, we identified PVT1, a long non-coding RNA, with the peptide HF10 derived from PVT1, presented by HLA-A24. This study aims to develop a single-chain variable fragment (scFv) that specifically recognizes the HLA-A24/HF10 complex (HF10 scFv) and to evaluate its specificity, reactivity, and therapeutic potential as part of a T cell engaging bispecific antibody (HF10xCD3) in vitro and in vivo. Using a scFv phage display library, we screened for scFv clones targeting the HLA-A24/HF10 peptide complex. The selected HF10 scFv was engineered into an IgG1 format (HF10-hIgG1), which demonstrated high affinity (KD = 2.18 × 10⁻⁸ M) and specific detection of the HLA-A24/HF10 complex on HLA-A24( +)/PVT1( +) tumor cell lines. Furthermore, HF10 scFv was incorporated into a T cell engaging bispecific antibody (HF10xCD3), which induced cytotoxicity in these tumor cell lines. In a mouse xenograft model, HF10xCD3 administration exhibited significant anti-tumor activity. In conclusion, HF10xCD3 represents a promising candidate for immunotherapy targeting solid tumors.https://doi.org/10.1007/s00262-025-03976-7Long non-coding RNAOsteosarcomaCarcinomaHLA-A24CTL epitopePeptide
spellingShingle Hirotaka Kato
Tomohide Tsukahara
Kenji Murata
Hiromu Nishikata
Yuka Mizue
Takashi Sasaya
Terufumi Kubo
Takayuki Kanaseki
Yoshihiko Hirohashi
Atsushi Oyagi
Tatsuo Maeda
Akihiro Miyazaki
Toshihiko Torigoe
Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
Cancer Immunology, Immunotherapy
Long non-coding RNA
Osteosarcoma
Carcinoma
HLA-A24
CTL epitope
Peptide
title Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
title_full Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
title_fullStr Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
title_full_unstemmed Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
title_short Development of a T cell engaging bispecific antibody targeting long non-coding RNA PVT1
title_sort development of a t cell engaging bispecific antibody targeting long non coding rna pvt1
topic Long non-coding RNA
Osteosarcoma
Carcinoma
HLA-A24
CTL epitope
Peptide
url https://doi.org/10.1007/s00262-025-03976-7
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