Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles

Abstract Despite the outstanding clinical success of immunotherapy, its therapeutic efficacy in glioblastoma (GBM) is still limited. To identify critical regulators of GBM immunity, we constructed a mouse single-guide RNA (sgRNA) library corresponding to all disease-related immune genes, and perform...

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Main Authors: Cheng Zou, Xiao Liu, Weizhong Wang, Lei He, Anan Yin, Zhengcong Cao, Maorong Zhu, Yuxin Wu, Xiaolin Liu, Jiying Ma, Yalong He, Shuning Wang, Wangqian Zhang, Wei Liu, Yingqi Zhang, Jintao Gu, Wei Lin, Kuo Zhang, Meng Li
Format: Article
Language:English
Published: BMC 2025-02-01
Series:Journal of Nanobiotechnology
Online Access:https://doi.org/10.1186/s12951-025-03182-8
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author Cheng Zou
Xiao Liu
Weizhong Wang
Lei He
Anan Yin
Zhengcong Cao
Maorong Zhu
Yuxin Wu
Xiaolin Liu
Jiying Ma
Yalong He
Shuning Wang
Wangqian Zhang
Wei Liu
Yingqi Zhang
Jintao Gu
Wei Lin
Kuo Zhang
Meng Li
author_facet Cheng Zou
Xiao Liu
Weizhong Wang
Lei He
Anan Yin
Zhengcong Cao
Maorong Zhu
Yuxin Wu
Xiaolin Liu
Jiying Ma
Yalong He
Shuning Wang
Wangqian Zhang
Wei Liu
Yingqi Zhang
Jintao Gu
Wei Lin
Kuo Zhang
Meng Li
author_sort Cheng Zou
collection DOAJ
description Abstract Despite the outstanding clinical success of immunotherapy, its therapeutic efficacy in glioblastoma (GBM) is still limited. To identify critical regulators of GBM immunity, we constructed a mouse single-guide RNA (sgRNA) library corresponding to all disease-related immune genes, and performed an in vivo CRISPR knockout (KO) screen in syngeneic GBM mouse models. We demonstrated that the deletion of GDF15 in GBM cells ameliorated the immunosuppressive tumor microenvironment (TME) and enhanced the antitumor efficacy of immune checkpoint blockade (ICB) response. Moreover, we designed unique nanoparticles for efficient encapsulation of CRISPR-Cas9, noninvasive brain delivery and tumor cell targeting, demonstrating an effective and safe strategy for GDF15 gene therapy. The CRISPR-Cas9 nanoparticles, known as ANPSS (Cas9/sgRNA), are easily created by enclosing a single Cas9/sgRNA complex in a polymer shell that is sensitive to glutathione. This shell also contains a dual-action ligand that aids in crossing the blood‒brain barrier, targeting tumor cells, and selectively releasing Cas9/sgRNA. Our encapsulating nanoparticles demonstrated promising GBM targeting, resulting in high GDF15 gene editing efficiency within brain tumors while showing minimal off-target gene editing in high-risk tissues. Treatment with ANPSS (Cas9/sgGDF15) effectively halted tumor growth, reversed immune suppression, and enhanced the efficacy of ICB therapy. These results emphasize the potential role of GDF15 in modulating the immune microenvironment and enhancing the effectiveness of current immunotherapy strategies for GBM. Graphical Abstract
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spelling doaj-art-fe7356da742f452fbc6a2b0f7a25a5592025-08-20T02:15:16ZengBMCJournal of Nanobiotechnology1477-31552025-02-0123111910.1186/s12951-025-03182-8Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticlesCheng Zou0Xiao Liu1Weizhong Wang2Lei He3Anan Yin4Zhengcong Cao5Maorong Zhu6Yuxin Wu7Xiaolin Liu8Jiying Ma9Yalong He10Shuning Wang11Wangqian Zhang12Wei Liu13Yingqi Zhang14Jintao Gu15Wei Lin16Kuo Zhang17Meng Li18State Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityDepartment of Neurosurgery, Xijing HospitalDepartment of Neurosurgery, Xijing HospitalState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityDepartment of Plastic and Reconstructive Surgery, Xijing Hospital, Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityDepartment of Neurosurgery, Xijing HospitalState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityDepartment of Neurosurgery, Xijing HospitalState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityDepartment of Neurosurgery, Xijing HospitalState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityState Key Laboratory of Cancer Biology, Biotechnology Center, School of Pharmacy, The Fourth Military Medical UniversityAbstract Despite the outstanding clinical success of immunotherapy, its therapeutic efficacy in glioblastoma (GBM) is still limited. To identify critical regulators of GBM immunity, we constructed a mouse single-guide RNA (sgRNA) library corresponding to all disease-related immune genes, and performed an in vivo CRISPR knockout (KO) screen in syngeneic GBM mouse models. We demonstrated that the deletion of GDF15 in GBM cells ameliorated the immunosuppressive tumor microenvironment (TME) and enhanced the antitumor efficacy of immune checkpoint blockade (ICB) response. Moreover, we designed unique nanoparticles for efficient encapsulation of CRISPR-Cas9, noninvasive brain delivery and tumor cell targeting, demonstrating an effective and safe strategy for GDF15 gene therapy. The CRISPR-Cas9 nanoparticles, known as ANPSS (Cas9/sgRNA), are easily created by enclosing a single Cas9/sgRNA complex in a polymer shell that is sensitive to glutathione. This shell also contains a dual-action ligand that aids in crossing the blood‒brain barrier, targeting tumor cells, and selectively releasing Cas9/sgRNA. Our encapsulating nanoparticles demonstrated promising GBM targeting, resulting in high GDF15 gene editing efficiency within brain tumors while showing minimal off-target gene editing in high-risk tissues. Treatment with ANPSS (Cas9/sgGDF15) effectively halted tumor growth, reversed immune suppression, and enhanced the efficacy of ICB therapy. These results emphasize the potential role of GDF15 in modulating the immune microenvironment and enhancing the effectiveness of current immunotherapy strategies for GBM. Graphical Abstracthttps://doi.org/10.1186/s12951-025-03182-8
spellingShingle Cheng Zou
Xiao Liu
Weizhong Wang
Lei He
Anan Yin
Zhengcong Cao
Maorong Zhu
Yuxin Wu
Xiaolin Liu
Jiying Ma
Yalong He
Shuning Wang
Wangqian Zhang
Wei Liu
Yingqi Zhang
Jintao Gu
Wei Lin
Kuo Zhang
Meng Li
Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
Journal of Nanobiotechnology
title Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
title_full Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
title_fullStr Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
title_full_unstemmed Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
title_short Targeting GDF15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment-responsive CRISPR-Cas9 nanoparticles
title_sort targeting gdf15 to enhance immunotherapy efficacy in glioblastoma through tumor microenvironment responsive crispr cas9 nanoparticles
url https://doi.org/10.1186/s12951-025-03182-8
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