CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo

Abstract Immune checkpoint (ICP) blockade has shown limited effectiveness in glioblastoma (GBM), particularly in the mesenchymal subtype, where interactions between immune cells and glioblastoma cancer stem cells (GSCs) drive immunosuppression and therapy resistance. Tailoring ICPs specific to GSCs...

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Main Authors: Nadia Rouatbi, Adam A. Walters, Alaa Zam, Yau Mun Lim, Alessia Marrocu, Revadee Liam‐Or, Joanne E. Anstee, James N. Arnold, Julie Tzu‐Wen Wang, Steven M. Pollard, Khuloud T. Al‐Jamal
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Advanced Science
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Online Access:https://doi.org/10.1002/advs.202407262
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author Nadia Rouatbi
Adam A. Walters
Alaa Zam
Yau Mun Lim
Alessia Marrocu
Revadee Liam‐Or
Joanne E. Anstee
James N. Arnold
Julie Tzu‐Wen Wang
Steven M. Pollard
Khuloud T. Al‐Jamal
author_facet Nadia Rouatbi
Adam A. Walters
Alaa Zam
Yau Mun Lim
Alessia Marrocu
Revadee Liam‐Or
Joanne E. Anstee
James N. Arnold
Julie Tzu‐Wen Wang
Steven M. Pollard
Khuloud T. Al‐Jamal
author_sort Nadia Rouatbi
collection DOAJ
description Abstract Immune checkpoint (ICP) blockade has shown limited effectiveness in glioblastoma (GBM), particularly in the mesenchymal subtype, where interactions between immune cells and glioblastoma cancer stem cells (GSCs) drive immunosuppression and therapy resistance. Tailoring ICPs specific to GSCs can enhance the antitumor immune response. This study proposes the use of lipid nanoparticles (LNPs) encapsulating CRISPR RNAs as an in vivo screening tool for ICPs in a syngeneic model of mesenchymal GSCs. Using PD‐L1 and CD47 to validate the proof of concept, intratumoral administration of LNPs in orthotopic tumors achieved efficient editing of ICPs, leading to enhanced immune cell infiltration within the tumor microenvironment. Targeting CD47 reduced tumor growth, suggesting improved cancer cell sensitization to the immune system post‐ICP editing. The study positions LNPs as a robust tool for in vivo validation of ICPs as therapeutic targets in clinically relevant GBM models. LNPs could serve as a screening tool in patient‐derived xenografts to identify and optimize ICP combinations, potentially expediting ICP translation and enhancing personalized GBM immunotherapies.
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spelling doaj-art-a68ea34b6ae347aba53e7fb6c6ecf7232025-08-20T01:49:41ZengWileyAdvanced Science2198-38442025-03-011212n/an/a10.1002/advs.202407262CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In VivoNadia Rouatbi0Adam A. Walters1Alaa Zam2Yau Mun Lim3Alessia Marrocu4Revadee Liam‐Or5Joanne E. Anstee6James N. Arnold7Julie Tzu‐Wen Wang8Steven M. Pollard9Khuloud T. Al‐Jamal10Institute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKComprehensive Cancer Centre Faculty of Life Sciences and Medicine King's College London, Guy's Hospital London SE1 1UL UKComprehensive Cancer Centre Faculty of Life Sciences and Medicine King's College London, Guy's Hospital London SE1 1UL UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKCentre for Regenerative Medicine Institute for Regeneration and Repair & Cancer Research UK Scotland Centre University of Edinburgh 5 Little France Drive Edinburgh EH16 4UU UKInstitute of Pharmaceutical Science Faculty of Life Sciences and Medicine King's College London Franklin‐Wilkins Building, 150 Stamford Street London SE1 9NH UKAbstract Immune checkpoint (ICP) blockade has shown limited effectiveness in glioblastoma (GBM), particularly in the mesenchymal subtype, where interactions between immune cells and glioblastoma cancer stem cells (GSCs) drive immunosuppression and therapy resistance. Tailoring ICPs specific to GSCs can enhance the antitumor immune response. This study proposes the use of lipid nanoparticles (LNPs) encapsulating CRISPR RNAs as an in vivo screening tool for ICPs in a syngeneic model of mesenchymal GSCs. Using PD‐L1 and CD47 to validate the proof of concept, intratumoral administration of LNPs in orthotopic tumors achieved efficient editing of ICPs, leading to enhanced immune cell infiltration within the tumor microenvironment. Targeting CD47 reduced tumor growth, suggesting improved cancer cell sensitization to the immune system post‐ICP editing. The study positions LNPs as a robust tool for in vivo validation of ICPs as therapeutic targets in clinically relevant GBM models. LNPs could serve as a screening tool in patient‐derived xenografts to identify and optimize ICP combinations, potentially expediting ICP translation and enhancing personalized GBM immunotherapies.https://doi.org/10.1002/advs.202407262CD47glioblastomaimmune checkpointimmunotherapynanoparticlesnucleic acid delivery
spellingShingle Nadia Rouatbi
Adam A. Walters
Alaa Zam
Yau Mun Lim
Alessia Marrocu
Revadee Liam‐Or
Joanne E. Anstee
James N. Arnold
Julie Tzu‐Wen Wang
Steven M. Pollard
Khuloud T. Al‐Jamal
CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
Advanced Science
CD47
glioblastoma
immune checkpoint
immunotherapy
nanoparticles
nucleic acid delivery
title CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
title_full CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
title_fullStr CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
title_full_unstemmed CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
title_short CD47 Knock‐Out Using CRISPR‐Cas9 RNA Lipid Nanocarriers Results in Reduced Mesenchymal Glioblastoma Growth In Vivo
title_sort cd47 knock out using crispr cas9 rna lipid nanocarriers results in reduced mesenchymal glioblastoma growth in vivo
topic CD47
glioblastoma
immune checkpoint
immunotherapy
nanoparticles
nucleic acid delivery
url https://doi.org/10.1002/advs.202407262
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