A self-cascade nanoCRISPR prompts transcellular penetration to potentiate gene editing and tumor killing
Chao Liu1, Yangsong Xu1, Ning Wang1
1Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
Abstract:
CRISPR/Cas9-based therapeutics face significant challenges in penetrating the dense microenvironment of solid tumors, resulting in insufficient gene editing and compromised treatment efficacy. Current nanostrategies, which mainly focus on the paracellular pathway attempted to improve gene editing performance, whereas their efficiency remains uneven in the heterogenous extracellular matrix. Here, the nanoCRISPR system is prepared with self-cascading mechanisms for gene editing-mediated robust apoptosis and transcellular penetration. NanoCRISPR unlocks its self-cascade capability within the matrix metallopeptidase 2-enriched tumor microenvironment, initiating the transcellular penetration. By facilitating cellular uptake, nanoCRISPR triggers robust apoptosis in edited malignancies, promoting further transcellular penetration and amplifying gene editing in neighboring tumor cells. Benefiting from self-cascade between robust apoptosis and transcellular penetration, nanoCRISPR demonstrates continuous gene transfection/tumor killing performance (transfection/apoptosis efficiency: 1st round: 85%/84.2%; 2nd round: 48%/27%) and homogeneous penetration. In xenograft tumor-bearing mice, nanoCRISPR treatment achieves remarkable anti-tumor efficacy (∼83%) and significant survival benefits with minimal toxicity. This strategy presents a promising paradigm emphasizing transcellular penetration to enhance the effectiveness of CRISPR-based antitumor therapeutics.
Insights
This study introduces nanoCRISPR, a novel system that enhances CRISPR/Cas9 gene editing in solid tumors by utilizing transcellular penetration. This approach overcomes tumor microenvironment barriers, improving therapeutic efficacy and reducing toxicity.
Area of Science:
- Biotechnology
- Nanomedicine
- Cancer Therapeutics
Background:
- CRISPR/Cas9 therapeutics struggle with solid tumor penetration, limiting gene editing efficiency and treatment outcomes.
- Existing nanostrategies targeting the paracellular pathway show limited and uneven efficacy due to heterogeneous tumor extracellular matrices.
Purpose of the Study:
- To develop a nanoCRISPR system with self-cascading mechanisms for enhanced transcellular penetration and gene editing in solid tumors.
- To improve CRISPR/Cas9-based cancer therapy effectiveness by overcoming tumor microenvironment barriers.
Main Methods:
- Engineered nanoCRISPR system with self-cascading properties activated by matrix metallopeptidase 2 (MMP2) in the tumor microenvironment.
- Utilized transcellular penetration initiated by nanoCRISPR for enhanced cellular uptake and gene editing.
- Demonstrated self-amplifying cycles of apoptosis and transcellular penetration for sustained gene editing and tumor cell killing.
Main Results:
- NanoCRISPR achieved efficient transcellular penetration and robust apoptosis in edited tumor cells.
- Demonstrated continuous gene transfection and tumor killing with high efficiency in sequential rounds (1st round: 85%/84.2%; 2nd round: 48%/27%).
- Achieved significant anti-tumor efficacy (∼83%) and survival benefits in xenograft mouse models with minimal toxicity.
Conclusions:
- The nanoCRISPR system effectively enhances CRISPR/Cas9 antitumor therapy through transcellular penetration and self-cascading apoptosis.
- This strategy offers a promising new paradigm for overcoming solid tumor barriers and improving gene editing-based cancer treatments.
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