[Construction and identification of a stable CT26 cell line expressing CD19-FLUC-GFP]

Area of Science:

• Oncology
• Immunotherapy
• Molecular Biology

Context:

• Solid tumors present challenges for chimeric antigen receptor T-cell (CAR-T) therapy due to a lack of specific targets.
• CD19 is a well-established target in hematological malignancies, but its utility in solid tumors is limited.
• Developing target-expressing solid tumor models is crucial for advancing CAR-T therapy.

Purpose:

• To construct a stable colon cancer cell line engineered to express CD19, firefly luciferase (FLUC), and green fluorescent protein (GFP).
• To evaluate the stability of CD19 expression and the susceptibility of the engineered cell line to CD19 CAR-T cell-mediated cytotoxicity.
• To establish a preclinical model for investigating CD19-targeted CAR-T cell therapy against solid tumors.

Summary:

• A stable CT26 colon cancer cell line (CT26-CD19-FLUC-GFP) was created using lentiviral transduction, consistently expressing CD19, FLUC, and GFP across multiple generations.
• Flow cytometry and mRNA analysis confirmed stable and high-level expression of CD19 and FLUC.
• CD19 CAR-T cells exhibited significant cytotoxicity against CT26-CD19-FLUC-GFP cells, and tumor growth was detectable via FLUC imaging in vivo.

Impact:

• Successfully established a validated, CD19-expressing colon cancer cell line model for preclinical CAR-T therapy research.
• Demonstrates the potential of engineering solid tumor cells to express specific targets for CAR-T cell recognition and elimination.
• Provides a valuable tool for further investigation into the efficacy and optimization of CD19-targeted CAR-T cell therapies for solid tumors.