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Targeting refractory diffuse large B cell lymphoma by CAR-WEE1 T-cells: In vitro evaluation
Hadeer Mohamed Ahmed1, Said Salama Moselhy1, Magda I Mohamad2
1Department of Biochemistry, Faculty of Science, Ain Shams University, Cairo, 11566, Egypt.
Abstract:
Refractory Diffuse Large B-cell Lymphoma (DLBCL) presents a major therapeutic challenge due to its resistance to standard treatments. Engineered T-cells, especially Chimeric Antigen Receptor (CAR) T-cells, have shown promise in overcoming drug resistance. This study investigates the effectiveness of WEE1-engineered T-cells in targeting and eliminating refractory DLBCL in vitro. CAR T-cells were created by transducing a 5th-generation CAR construct designed to recognize WEE1, a surface antigen commonly found on refractory DLBCL cells. The cytotoxic effect of engineered T-cells was tested against Rituximab-resistant DLBCL cells (RR-NU-DUL-1). Apoptosis and cell cycle were evaluated using flow cytometry. Quantitative Real-time PCR (RT-PCR) was used to measure the expression of WEE1, BCL2, and CDK2. The results showed a significant increase in target cell lysis, apoptosis, and necrosis, a significant reduction in the percentage of cells in the G2M phase of the cell cycle, as well as a decrease in gene expression level, indicating strong anti-tumor activity. These findings suggest that CAR T-cell therapy holds great promise for treating refractory DLBCL, offering a potential path for clinical application. This in vitro evaluation highlights the potential of WEE1-engineered T-cells as a targeted treatment strategy for refractory DLBCL, emphasizing their clinical applicability and ability to overcome resistance mechanisms in this aggressive lymphoma subtype.
Insights
Engineered T-cells targeting WEE1 show promise against refractory Diffuse Large B-cell Lymphoma (DLBCL). This therapy effectively eliminated DLBCL cells in vitro, suggesting a new treatment avenue.
Area of Science:
- Oncology
- Immunotherapy
- Cell Biology
Background:
- Refractory Diffuse Large B-cell Lymphoma (DLBCL) poses significant treatment challenges due to resistance.
- Engineered T-cells, particularly Chimeric Antigen Receptor (CAR) T-cells, offer potential for overcoming drug resistance.
Purpose of the Study:
- To investigate the efficacy of WEE1-engineered T-cells against refractory DLBCL in vitro.
- To assess the ability of these engineered T-cells to target and eliminate DLBCL cells resistant to standard therapies.
Main Methods:
- Development of 5th-generation CAR T-cells targeting the WEE1 antigen on DLBCL cells.
- In vitro evaluation of engineered T-cell cytotoxicity against Rituximab-resistant DLBCL cell lines.
- Analysis of apoptosis, cell cycle, and gene expression (WEE1, BCL2, CDK2) via flow cytometry and RT-PCR.
Main Results:
- Significant increase in DLBCL cell lysis, apoptosis, and necrosis mediated by WEE1-engineered T-cells.
- Demonstrated reduction in the G2M phase of the cell cycle in target cells.
- Observed decrease in WEE1, BCL2, and CDK2 gene expression, indicating potent anti-tumor activity.
Conclusions:
- WEE1-engineered CAR T-cell therapy exhibits strong anti-tumor activity against refractory DLBCL in vitro.
- This approach shows potential for overcoming resistance mechanisms in aggressive lymphoma subtypes.
- Findings support the clinical applicability of WEE1-targeted CAR T-cell therapy for refractory DLBCL.
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