GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy

Abstract The lack of targetable antigens poses a significant challenge in developing effective cancer‐targeted therapies. Cell surface translocation of endoplasmic reticulum (ER) chaperones, such as glucose‐regulated protein 78 (GRP78), during malignancy, drug resistance, and ER stress induced by th...

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Main Authors: Huifang Wang, Runhua Zhou, Chengchao Xu, Lingyun Dai, Rui Hou, Liuhai Zheng, Chunjin Fu, Guangwei Shi, Jingwei Wang, Yang Li, Jinpeng Cen, Xiaolong Xu, Le Yu, Yilei Li, Jigang Wang, Qingfeng Du, Zhijie Li
Format: Article
Language:English
Published: Wiley 2025-05-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202408086
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author Huifang Wang
Runhua Zhou
Chengchao Xu
Lingyun Dai
Rui Hou
Liuhai Zheng
Chunjin Fu
Guangwei Shi
Jingwei Wang
Yang Li
Jinpeng Cen
Xiaolong Xu
Le Yu
Yilei Li
Jigang Wang
Qingfeng Du
Zhijie Li
author_facet Huifang Wang
Runhua Zhou
Chengchao Xu
Lingyun Dai
Rui Hou
Liuhai Zheng
Chunjin Fu
Guangwei Shi
Jingwei Wang
Yang Li
Jinpeng Cen
Xiaolong Xu
Le Yu
Yilei Li
Jigang Wang
Qingfeng Du
Zhijie Li
author_sort Huifang Wang
collection DOAJ
description Abstract The lack of targetable antigens poses a significant challenge in developing effective cancer‐targeted therapies. Cell surface translocation of endoplasmic reticulum (ER) chaperones, such as glucose‐regulated protein 78 (GRP78), during malignancy, drug resistance, and ER stress induced by therapies, offers a promising pan‐cancer target. To target GRP78, nanobody C5, identified from a phage library and exhibiting high affinity for human and mouse GRP78, is utilized to develop the Pseudomonas exotoxin (PE) immunotoxin C5‐PE38. C5‐PE38 induced ER stress, apoptosis and immunogenic cell death in targeted cells and showed antitumor efficacy against colorectal cancer and melanoma models without obvious toxicity. Mechanistically, transcriptome profiling showed that C5‐PE38 reshaped the tumor immune microenvironment with enhanced innate and adaptive immune response and response to interferon beta. Moreover, C5‐PE38‐induced cell death could trans‐activate STING pathway in dendritic cells and macrophages, promoting CD8+ T cell infiltration. It also sensitizes both primary and metastatic melanomas to anti‐PD1 therapy, partly through STING activation. Overall, this study unveils a feasible GRP78 nanobody‐directed therapy strategy for single or combinatorial cancer intervention. This work finds that C5‐PE38‐induced cell death stimulates STING‐dependent cytosolic DNA release to promote antitumor immunity, a mechanism not previously reported for PE38, providing valuable insights for its clinical use.
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spelling doaj-art-45b5d98db04d4372ab2bcd47d05dc1112025-08-20T03:13:30ZengWileyAdvanced Science2198-38442025-05-011219n/an/a10.1002/advs.202408086GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor ImmunotherapyHuifang Wang0Runhua Zhou1Chengchao Xu2Lingyun Dai3Rui Hou4Liuhai Zheng5Chunjin Fu6Guangwei Shi7Jingwei Wang8Yang Li9Jinpeng Cen10Xiaolong Xu11Le Yu12Yilei Li13Jigang Wang14Qingfeng Du15Zhijie Li16Department of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaClinical Pharmacy Center Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Neurosurgery & Medical Research Center Shunde Hospital Southern Medical University (The First People's Hospital of Shunde Foshan) Guangzhou 510515 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaDepartment of Urology Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaClinical Pharmacy Center Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 ChinaClinical Pharmacy Center Nanfang Hospital Southern Medical University Guangzhou Guangdong 510515 ChinaSchool of Traditional Chinese Medicine and School of Pharmaceutical Sciences Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical Sciences Southern Medical University Guangzhou Guangdong 510515 ChinaSchool of Traditional Chinese Medicine and School of Pharmaceutical Sciences Guangdong Provincial Key Laboratory of New Drug Screening School of Pharmaceutical Sciences Southern Medical University Guangzhou Guangdong 510515 ChinaDepartment of Critical Care Medicine Guangdong Provincial Clinical Research Center for Geriatrics Shenzhen Clinical Research Centre for Geriatrics Department of Nuclear Medicine Shenzhen People's Hospital (The First Affiliated Hospital, Southern University of Science and Technology; The Second Clinical Medical CollegeJinan University) Shenzhen Guangdong 518020 ChinaAbstract The lack of targetable antigens poses a significant challenge in developing effective cancer‐targeted therapies. Cell surface translocation of endoplasmic reticulum (ER) chaperones, such as glucose‐regulated protein 78 (GRP78), during malignancy, drug resistance, and ER stress induced by therapies, offers a promising pan‐cancer target. To target GRP78, nanobody C5, identified from a phage library and exhibiting high affinity for human and mouse GRP78, is utilized to develop the Pseudomonas exotoxin (PE) immunotoxin C5‐PE38. C5‐PE38 induced ER stress, apoptosis and immunogenic cell death in targeted cells and showed antitumor efficacy against colorectal cancer and melanoma models without obvious toxicity. Mechanistically, transcriptome profiling showed that C5‐PE38 reshaped the tumor immune microenvironment with enhanced innate and adaptive immune response and response to interferon beta. Moreover, C5‐PE38‐induced cell death could trans‐activate STING pathway in dendritic cells and macrophages, promoting CD8+ T cell infiltration. It also sensitizes both primary and metastatic melanomas to anti‐PD1 therapy, partly through STING activation. Overall, this study unveils a feasible GRP78 nanobody‐directed therapy strategy for single or combinatorial cancer intervention. This work finds that C5‐PE38‐induced cell death stimulates STING‐dependent cytosolic DNA release to promote antitumor immunity, a mechanism not previously reported for PE38, providing valuable insights for its clinical use.https://doi.org/10.1002/advs.202408086cell surface glucose‐regulated protein 78 (GRP78)immunotherapyimmunotoxinSTING pathwaytargeted therapy
spellingShingle Huifang Wang
Runhua Zhou
Chengchao Xu
Lingyun Dai
Rui Hou
Liuhai Zheng
Chunjin Fu
Guangwei Shi
Jingwei Wang
Yang Li
Jinpeng Cen
Xiaolong Xu
Le Yu
Yilei Li
Jigang Wang
Qingfeng Du
Zhijie Li
GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
Advanced Science
cell surface glucose‐regulated protein 78 (GRP78)
immunotherapy
immunotoxin
STING pathway
targeted therapy
title GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
title_full GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
title_fullStr GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
title_full_unstemmed GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
title_short GRP78 Nanobody‐Directed Immunotoxin Activates Innate Immunity Through STING Pathway to Synergize Tumor Immunotherapy
title_sort grp78 nanobody directed immunotoxin activates innate immunity through sting pathway to synergize tumor immunotherapy
topic cell surface glucose‐regulated protein 78 (GRP78)
immunotherapy
immunotoxin
STING pathway
targeted therapy
url https://doi.org/10.1002/advs.202408086
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