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Targeting programmed cell death ligand 1 by CRISPR/Cas9 in osteosarcoma cells
Yunfei Liao1,2, Lulu Chen1, Yong Feng2,3
1Department of Endocrinology, Wuhan Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Abstract:
Programmed cell death ligand 1 (PD-L1) is a transmembrane protein that is expressed on tumor cells that suppresses the T cell-mediated immune response. Therapies targeting the PD-L1 pathway promote anti-tumor immunity and have shown promising results in some types of cancers. However, the functional and therapeutic roles of PD-L1 in osteosarcoma remain largely unknown. In this study, we found that PD-L1 protein was expressed in osteosarcoma cell lines and tissue microarray of patient tumors. Tissue microarray immunohistochemistry analysis showed that the overall and five-year survival rates of patients with high levels of PD-L1 expression were significantly shorter than patients with low levels. High levels of PD-L1 expression were also associated with metastasis in osteosarcoma patients. Furthermore, we applied the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system to target PD-L1 gene at the DNA level in osteosarcoma cell lines. We found that the expression of PD-L1 could be efficiently disrupted by CRISPR/Cas9 system and PD-L1 knockdown increased drug sensitivities for doxorubicin and paclitaxel. These results suggest that PD-L1 is an independent prognostic factor in osteosarcoma and that PD-L1 knockout by CRISPR/Cas9 may be a therapeutic approach for the treatment of osteosarcoma.
Insights
Programmed cell death ligand 1 (PD-L1) is expressed in osteosarcoma, correlating with poorer survival and metastasis. Targeting PD-L1 with CRISPR/Cas9 enhances chemotherapy sensitivity, suggesting a novel therapeutic strategy.
Area of Science:
- Oncology
- Immunology
- Genetics
Background:
- Programmed cell death ligand 1 (PD-L1) is a transmembrane protein that inhibits anti-tumor immune responses.
- PD-L1 targeted therapies show promise in various cancers, but its role in osteosarcoma is unclear.
- Understanding PD-L1's function is crucial for developing new osteosarcoma treatments.
Purpose of the Study:
- To investigate the expression and prognostic significance of PD-L1 in osteosarcoma.
- To explore the therapeutic potential of targeting PD-L1 using CRISPR/Cas9 gene editing in osteosarcoma.
Main Methods:
- Immunohistochemistry on osteosarcoma tissue microarrays to assess PD-L1 expression.
- CRISPR/Cas9 gene editing to knock out the PD-L1 gene in osteosarcoma cell lines.
- Evaluation of drug sensitivity to doxorubicin and paclitaxel following PD-L1 knockdown.
Main Results:
- PD-L1 protein is expressed in osteosarcoma cell lines and patient tumors.
- High PD-L1 expression correlates with significantly shorter overall and five-year survival rates.
- High PD-L1 levels are associated with increased metastasis in osteosarcoma patients.
- CRISPR/Cas9 efficiently disrupted PD-L1 expression, enhancing sensitivity to doxorubicin and paclitaxel.
Conclusions:
- PD-L1 serves as an independent prognostic factor in osteosarcoma.
- PD-L1 knockout via CRISPR/Cas9 presents a potential therapeutic strategy for osteosarcoma treatment.
- Targeting PD-L1 may improve the efficacy of existing chemotherapies in osteosarcoma.