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Cdk5 knocking out mediated by CRISPR-Cas9 genome editing for PD-L1 attenuation and enhanced antitumor immunity
Huan Deng1, Songwei Tan1, Xueqin Gao1
1School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Abstract:
Blocking the programmed death-ligand 1 (PD-L1) on tumor cells with monoclonal antibody therapy has emerged as powerful weapon in cancer immunotherapy. However, only a minority of patients presented immune responses in clinical trials. To develop an alternative treatment method based on immune checkpoint blockade, we designed a novel and efficient CRISPR-Cas9 genome editing system delivered by cationic copolymer aPBAE to downregulate PD-L1 expression on tumor cells via specifically knocking out Cyclin-dependent kinase 5 (Cdk5) gene in vivo. The expression of PD-L1 on tumor cells was significantly attenuated by knocking out Cdk5, leading to effective tumor growth inhibition in murine melanoma and lung metastasis suppression in triple-negative breast cancer. Importantly, we demonstrated that aPBAE/Cas9-Cdk5 treatment elicited strong T cell-mediated immune responses in tumor microenvironment that the population of CD8+ T cells was significantly increased while regulatory T cells (Tregs) was decreased. It may be the first case to exhibit direct in vivo PD-L1 downregulation via CRISPR-Cas9 genome editing technology for cancer therapy. It will provide promising strategy for preclinical antitumor treatment through the combination of nanotechnology and genome engineering.
Insights
This study introduces a novel CRISPR-Cas9 gene editing system to reduce PD-L1 expression in cancer cells by targeting the Cdk5 gene. This approach enhances anti-tumor immunity and shows promise for cancer immunotherapy.
Area of Science:
- Oncology
- Immunotherapy
- Gene Editing
- Nanotechnology
Background:
- Monoclonal antibody therapy targeting PD-L1 is a key cancer immunotherapy, but patient response rates are limited.
- Developing alternative immune checkpoint blockade strategies is crucial for improving cancer treatment outcomes.
Purpose of the Study:
- To design and evaluate a novel CRISPR-Cas9 genome editing system for *in vivo* downregulation of PD-L1 expression on tumor cells.
- To investigate the potential of targeting the Cyclin-dependent kinase 5 (Cdk5) gene for cancer therapy.
Main Methods:
- A CRISPR-Cas9 genome editing system was developed and delivered using a cationic copolymer (aPBAE).
- The system was designed to specifically knock out the Cdk5 gene *in vivo* to reduce PD-L1 expression.
- The efficacy was tested in murine melanoma and triple-negative breast cancer models.
Main Results:
- Knocking out the Cdk5 gene significantly attenuated PD-L1 expression on tumor cells.
- The treatment led to effective tumor growth inhibition in melanoma and suppressed lung metastasis in breast cancer.
- The aPBAE/Cas9-Cdk5 treatment significantly increased CD8+ T cells and decreased regulatory T cells (Tregs) in the tumor microenvironment, indicating enhanced T cell-mediated immunity.
Conclusions:
- This study presents the first demonstration of direct *in vivo* PD-L1 downregulation using CRISPR-Cas9 genome editing for cancer therapy.
- The combination of nanotechnology and genome engineering offers a promising preclinical strategy for cancer treatment by modulating the tumor immune microenvironment.
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