¹²⁴I-labelled BMSC-Derived Extracellular Vesicles Deliver CRISPR/Cas9 Ribonucleoproteins With a GFP-Reporter System to Inhibit Osteosarcoma Proliferation and Metastasis

Yujie Pan ¹, Xianteng Yang ², Zhirui Zeng ³

⁰Department of Trauma Centre, The Affiliated Hospital of Guizhou Medical University, Guiyang, Guizhou, China.

Journal of Extracellular Vesicles +

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July 17, 2025

View abstract on PubMed

Summary

Targeting KCNJ2 with CRISPR/Cas9 delivered by engineered extracellular vesicles (EVs) effectively suppresses osteosarcoma metastasis. This novel therapy disrupts a key regulatory loop, inhibiting tumor growth and improving patient prognosis.

Area Of Science

Oncology
Molecular Biology
Biotechnology

Background

Osteosarcoma metastasis is a major cause of poor prognosis.
Hypoxia promotes osteosarcoma progression via HIF-1α.
A positive feedback loop between KCNJ2 and HIF-1α drives osteosarcoma proliferation and metastasis.

Purpose Of The Study

To develop a KCNJ2-targeted therapy to disrupt the KCNJ2-HIF-1α feedback loop.
To utilize CRISPR/Cas9 gene editing technology for osteosarcoma treatment.
To establish an efficient and targeted delivery system for CRISPR/Cas9 using engineered extracellular vesicles (EVs).

Main Methods

Developed a fluorescence-based reporter system to confirm CRISPR/Cas9 editing efficiency.
Engineered radiolabeled EVs from bone marrow mesenchymal stem cells (BMSCs) to deliver CRISPR/Cas9 targeting KCNJ2.
Validated in vitro gene editing and in vivo targeted distribution of engineered EVs.

Main Results

Confirmed efficient in vitro gene editing of KCNJ2 using the reporter system.
Demonstrated targeted in vivo distribution of radiolabeled EVs.
Showed that <sup>124</sup>I@EVs-Cas9 effectively suppresses osteosarcoma proliferation and metastasis by inhibiting KCNJ2 and promoting HIF-1α degradation.

Conclusions

Engineered EVs are effective carriers for CRISPR/Cas9 delivery in osteosarcoma.
Targeting the KCNJ2-HIF-1α feedback loop with CRISPR/Cas9 offers a promising therapeutic strategy.
This approach significantly inhibits osteosarcoma progression and metastasis.

Keywords:

CRISPR‐Cas9 bone marrow mesenchymal stem cells extracellular vesicles osteosarcoma