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Cited by 132CRISPR and Genetic EngineeringExtracellular vesicles in diseaseRNA Interference and Gene Delivery

Engineered Extracellular Vesicle-Delivered CRISPR/Cas9 for Radiotherapy Sensitization of Glioblastoma

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Liu, Xiao, Cao, Zhengcong, Wang, Weizhong, Zou, Cheng, Wang, Yingwen, Pan, Luxiang, Jia, Bo, Zhang, Kuo +9 more·2023·ACS Nano
DOI 10.1021/acsnano.2c12857PMC PMC10510715
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Abstract

loss-of-function genome-wide CRISPR screen was carried out in orthotopic tumors in mice subjected to radiation treatment to identify synthetic lethal genes associated with radiotherapy. Using functional screening and transcriptome analyses, glutathione synthetase (GSS) was found to be a potential regulator of radioresistance through ferroptosis. High GSS levels were closely related to poor prognosis and relapse in patients with glioma. Mechanistic studies demonstrated that GSS was associated with the suppression of radiotherapy-induced ferroptosis in glioma cells. The depletion of GSS resulted in the disruption of glutathione (GSH) synthesis, thereby causing the inactivation of GPX4 and iron accumulation, thus enhancing the induction of ferroptosis upon radiotherapy treatment. Moreover, to overcome the obstacles to broad therapeutic translation of CRISPR editing, we report a previously unidentified genome editing delivery system, in which Cas9 protein/sgRNA complex was loaded into Angiopep-2 (Ang) and the trans-activator of the transcription (TAT) peptide dual-modified extracellular vesicle (EV), which not only targeted the blood-brain barrier (BBB) and GBM but also permeated the BBB and penetrated the tumor. Our encapsulating EVs showed encouraging signs of GBM tissue targeting, which resulted in high GSS gene editing efficiency in GBM (up to 67.2%) with negligible off-target gene editing. These results demonstrate that a combination of unbiased genetic screens, and CRISPR-Cas9-based gene therapy is feasible for identifying potential synthetic lethal genes and, by extension, therapeutic targets.

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  • Even with nanotechnology, the efficiency of targeting disease or disordered area cells with EVs is quite low.

    “To treat CNS disorders using EVs, EVs should only target the disease or disordered area cells, but even with nanotechnology, efficiency is quite low [333–336].”

    From Extracellular vesicles as nanotheranostic platforms for targeted neurological disorder interventions (2024) · cites this paper as [335]

    The Results section reports weak performance for unmodified EVs: they remained near the surface of glioblastoma organoids, and the measured gene-editing frequency was 2.5%. But it also reports better targeting with peptide-modified EVs, while the Discussion says the engineered EVs efficiently targeted glioma. The paper therefore supports a limitation of unmodified EVs, not the broader claim that targeting remains quite inefficient even with nanotechnology. No page numbers are available in our copy. (LLM, conf=0.84)

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References 42 · Cited by 132

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Authors
Liu, X., Cao, Z., Wang, W., Zou, C., Wang, Y., Pan, L., Jia, B., Zhang, K., Zhang, W., Li, W., Hao, Q., Zhang, Y., Zhang, W., Xue, X., Lin, W., Li, M., & Gu, J.
Affiliations
  • Xijing Hospital (CN)
  • State Key Laboratory of Cancer Biology (CN)
  • Shaanxi Normal University (CN)
Corresponding author
  • Lin, Wei
  • Li, Meng
  • Gu, Jintao
ORCID
Venue
ACS Nano
Publisher
American Chemical Society (ACS)
Published
2023-08-30
Volume
17
Issue
17
Pages
16432-16447
ISSN
  • 1936-0851 (ISSN-L)
  • 1936-086X
DOI
10.1021/acsnano.2c12857
PMC
PMC10510715
OpenAlex
W4386271869
Cited by
132
References
42
Citations by year
2026: 42 · 2025: 55 · 2024: 33 · 2023: 2
FWCI
11.76
Type
journal article
Language
en
Open access
Licence
Locations
  • ACS Nano (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
National Natural Science Foundation of China (81803053, 81872049, 82003220)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
Keywords
glioblastoma, radiosensitization, CRISPR/Cas9 gene therapy, extracellular vesicles, Angiopep-2, TAT peptide, blood-brain barrier targeting, glutathione synthetase, ferroptosis, GPX4, glutathione synthesis, radioresistance (biology), synthetic lethality, genome-wide CRISPR screen, orthotopic glioma model
MeSH terms
Extracellular Vesicles, RNA, Guide, CRISPR-Cas Systems, Animals, Glioblastoma, Glioma, Glutathione, Mice, CRISPR-Cas Systems

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