In Vivo CRISPR Screen Identifies RPS19 as an Immunotherapy Target in Hepatocellular Carcinoma
5 affiliations
- 1State Key Laboratory of Holistic Integrative Management of Gastrointestinal Cancers Department of Biopharmaceutics School of Pharmacy Fourth Military Medical University Xi'an China
- 2Department of Neurosurgery General Hospital of Central Theater Command Wuhan China
- 3Department of Neurosurgery Tangdu Hospital Fourth Military Medical University Xi'an China
- 4Department of Experiment Surgery Xijing Hospital Fourth Military Medical University Xi'an China
- 5Department of Oncology 940th Hospital Joint Logistic Support Force Lanzhou China
Abstract
Despite the remarkable clinical success of immunotherapy, its efficacy against hepatocellular carcinoma (HCC) remains severely limited, highlighting an urgent need for novel targets and strategies to overcome immune evasion. Here, we aimed to identify key regulators of HCC immunity via unbiased functional screening and develop a targeted therapy to remodel the immunosuppressive tumor microenvironment (TME). We performed an in vivo CRISPR‐knockout screen using a 4500‐gene sgRNA library in syngeneic HCC mouse models. Single‐cell RNA sequencing of human HCC tissues and in vivo macrophage‐depletion assays were utilized to dissect cellular populations and validate underlying mechanisms. We identified ribosomal protein S19 (RPS19) as a central mediator of HCC immune evasion, whose deletion in HCC cells mitigates the immunosuppressive TME. Mechanistically, scRNA‐seq pinpointed tumor‐associated macrophages as the key effector subset, whereby complement C5a receptor 1 (C5aR1) transduces RPS19 signals to trigger macrophage‐dependent immune suppression. We further constructed SP94‐peptide‐modified disulfide‐bond‐containing nanoparticles (SNPSS) for targeted CRISPR‐Cas9 delivery. SNPSS efficiently homes to HCC cells, releases payloads within the glutathione‐rich TME, downregulates RPS19, reverses immune suppression, and robustly suppresses HCC progression in vivo. Collectively, the RPS19‐C5aR1‐macrophage axis fuels HCC immune escape, and SNPSS‐based RPS19 gene‐editing offers a promising strategy to remodel the TME and boost HCC immunotherapy efficacy.
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BibTeX
@article{he2026vivo,
title = {{In Vivo CRISPR Screen Identifies RPS19 as an Immunotherapy Target in Hepatocellular Carcinoma}},
author = {Lei He and Duo Yu and Fei Xie and Kaisheng Hou and Zhaozhao Li and Meifang Dai and Jingyi Yun and Haiyan Cao and Yang Xiao and Yueyuan Qiu and Gege Shi and Wangqian Zhang and Shuning Wang and Qiang Hao and Jieyu Zhang and Zhaowei Wang and Yingqi Zhang and Kuo Zhang and Jintao Gu and Haichen Du and Meng Li},
journal = {Advanced Science},
year = {2026},
month = oct,
doi = {10.1002/advs.78430},
url = {https://doi.org/10.1002/advs.78430},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - In Vivo CRISPR Screen Identifies RPS19 as an Immunotherapy Target in Hepatocellular Carcinoma
AU - Lei He
AU - Duo Yu
AU - Fei Xie
AU - Kaisheng Hou
AU - Zhaozhao Li
AU - Meifang Dai
AU - Jingyi Yun
AU - Haiyan Cao
AU - Yang Xiao
AU - Yueyuan Qiu
AU - Gege Shi
AU - Wangqian Zhang
AU - Shuning Wang
AU - Qiang Hao
AU - Jieyu Zhang
AU - Zhaowei Wang
AU - Yingqi Zhang
AU - Kuo Zhang
AU - Jintao Gu
AU - Haichen Du
AU - Meng Li
PY - 2026
DA - 2026/10/25
JO - Advanced Science
DO - 10.1002/advs.78430
UR - https://doi.org/10.1002/advs.78430
AB - Despite the remarkable clinical success of immunotherapy, its efficacy against hepatocellular carcinoma (HCC) remains severely limited, highlighting an urgent need for novel targets and strategies to overcome immune evasion. Here, we aimed to identify key regulators of HCC immunity via unbiased functional screening and develop a targeted therapy to remodel the immunosuppressive tumor microenvironment (TME). We performed an in vivo CRISPR‐knockout screen using a 4500‐gene sgRNA library in syngeneic HCC mouse models. Single‐cell RNA sequencing of human HCC tissues and in vivo macrophage‐depletion assays were utilized to dissect cellular populations and validate underlying mechanisms. We identified ribosomal protein S19 (RPS19) as a central mediator of HCC immune evasion, whose deletion in HCC cells mitigates the immunosuppressive TME. Mechanistically, scRNA‐seq pinpointed tumor‐associated macrophages as the key effector subset, whereby complement C5a receptor 1 (C5aR1) transduces RPS19 signals to trigger macrophage‐dependent immune suppression. We further constructed SP94‐peptide‐modified disulfide‐bond‐containing nanoparticles (SNPSS) for targeted CRISPR‐Cas9 delivery. SNPSS efficiently homes to HCC cells, releases payloads within the glutathione‐rich TME, downregulates RPS19, reverses immune suppression, and robustly suppresses HCC progression in vivo. Collectively, the RPS19‐C5aR1‐macrophage axis fuels HCC immune escape, and SNPSS‐based RPS19 gene‐editing offers a promising strategy to remodel the TME and boost HCC immunotherapy efficacy.
SN - 2198-3844
ER -