Kordomalarda PD-L1/PDGFR-α hedefli kimerik antijen reseptörü taşıyan doğal katil(PD-L1/PDGFR-α/CAR-NK) hücre terapisi

Chordoma, a rare neoplasm, is a type of bone tumor that mostly appear in the axial skeleton. The Food and Drug Administration (FDA)-approved chemotherapeutics for chordoma treatment is unavailable. Conventional treatment options includen-blococ resection or radiation therapy. Additionally, receptor tyrosine kinase (RTK) inhibitors are widely recommended. Therefore, a targeting strategy against RTKs by chimeric antigen receptors is promising. Growing interest also evaluates checkpoint inhibitors in chordoma and several studies yielded encouraging outcomes both ithe n pre-clinical and clinical stages. Chimeric antigen receptor (CAR) fused cytotoxic cells fulfilling components of immunotherapeutic applications. However, both in the bench side and clinical utilization, benefits are restricted by the challenges in design and transfer to T and natural killer cells. Target, promoter or ,co-stimulatory domain selections lead the fate of chimeric antigen receptor-mediated cytotoxic function. Still, a comprehensive investigation of basic CAR structure and designing the structure accordinly to research outcomes does not guarantee the functionality. Therefore, each individual CAR structure requires its own optimization, and pre-clinical evaluation. In this thesis, a PD-L1 and PDGFR-α targeting combinatorial CAR aimed to be designed for chordoma treatment. The PD-L1/PDGFR-α CAR structure cloned into lentiviral or sleeping beauty transposon backbones. The expressibility of the designed CAR structure demonstrated with "easy-to-transfect/transduct" cell lines (Human immortalized astrocytes and HEK293T cells) to avoid labor intensive evaluation. Later, transfection and transduction optimizations were performed in natural killer cell lines for the transfer of CAR vectors. Following CAR vector transfer expression analysis within the cell and cell surface was evaluated. Our findings demontrade that vector size, transfection/transduction methods, choice of cell line and the design itself has profound effects on CAR-NK cell generation.