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Updated: Jun 29, 2025
Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
Published on: May 30, 2025
Cryo-shocked tumor cells deliver CRISPR-Cas9 for lung cancer regression by synthetic lethality
Feng Liu1,2, Minhang Xin1, Huiheng Feng1,2
1National Key Laboratory of Advanced Drug Delivery and Release Systems, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
Abstract:
Although CRISPR-mediated genome editing holds promise for cancer therapy, inadequate tumor targeting and potential off-target side effects hamper its outcomes. In this study, we present a strategy using cryo-shocked lung tumor cells as a CRISPR-Cas9 delivery system for cyclin-dependent kinase 4 (CDK4) gene editing, which initiates synthetic lethal in KRAS-mutant non-small cell lung cancer (NSCLC). By rapidly liquid nitrogen shocking, we effectively eliminate the pathogenicity of tumor cells while preserving their structure and surface receptor activity. This delivery system enables the loaded CRISPR-Cas9 to efficiently target to lung through the capture in pulmonary capillaries and interactions with endothelial cells. In a NSCLC-bearing mouse model, the drug accumulation is increased nearly fourfold in lung, and intratumoral CDK4 expression is substantially down-regulated compared to CRISPR-Cas9 lipofectamine nanoparticles administration. Furthermore, CRISPR-Cas9 editing-mediated CDK4 ablation triggers synthetic lethal in KRAS-mutant NSCLC and prolongs the survival of mice.
Insights
Cryo-shocked tumor cells deliver CRISPR-Cas9 to edit the cyclin-dependent kinase 4 (CDK4) gene in KRAS-mutant lung cancer. This novel delivery system enhances tumor targeting and triggers synthetic lethality, improving survival in mice.
Area of Science:
- Oncology
- Gene Therapy
- Nanomedicine
Background:
- CRISPR-Cas9 gene editing shows potential for cancer therapy but faces challenges in tumor targeting and off-target effects.
- KRAS-mutant non-small cell lung cancer (NSCLC) is a significant clinical challenge requiring novel therapeutic strategies.
Purpose of the Study:
- To develop a novel delivery system for CRISPR-Cas9 to target lung tumors and induce synthetic lethality in KRAS-mutant NSCLC.
- To evaluate the efficacy of cryo-shocked tumor cells as a delivery vehicle for CRISPR-Cas9 mediated CDK4 gene editing.
Main Methods:
- Lung tumor cells were cryo-shocked using liquid nitrogen to eliminate pathogenicity while preserving structural and surface receptor integrity.
- The cryo-shocked cells were loaded with CRISPR-Cas9 targeting the cyclin-dependent kinase 4 (CDK4) gene.
- The delivery system's efficacy was assessed in a NSCLC mouse model, comparing lung drug accumulation and CDK4 downregulation against lipofectamine nanoparticles.
Main Results:
- Cryo-shocked tumor cells effectively delivered CRISPR-Cas9 to lung tumors, with a fourfold increase in lung drug accumulation compared to lipofectamine nanoparticles.
- Intratumoral CDK4 expression was significantly downregulated following treatment with the engineered tumor cell delivery system.
- CRISPR-Cas9 mediated CDK4 ablation induced synthetic lethality in KRAS-mutant NSCLC, leading to prolonged survival in treated mice.
Conclusions:
- Cryo-shocked tumor cells represent a promising and effective delivery system for CRISPR-Cas9 gene editing in NSCLC.
- This strategy overcomes tumor targeting limitations and off-target effects, offering a new therapeutic avenue for KRAS-mutant lung cancer.
- The induction of synthetic lethality via CDK4 gene editing demonstrates significant therapeutic potential and improved survival outcomes.
