2002 NY-ESO-1-Specific TCR-T Cells Effectively Targets Glioblastoma Cancer Stem Cells

INTRODUCTION: Glioblastoma (GBM) recurrence remains an unmet medical challenge, largely attributed to resilient glioma cancer stem cells (CSCs) that evade conventional therapies. TCR T cells bear virally infected T cell receptors for targeting antigen-bearing tumor cells. NY-ESO-1-specific T cell receptor (TCR) T cells can target GBM cells. Decitabine (DAC) increases NY-ESO-1 expression, specifically in CSCs, potentially enabling the eradication of this cell population. METHODS: NY-ESO-1 expression was analyzed by immunocytochemistry and RT-PCR. Cytotoxicity assays with T cells and DAC pre-treated tumor cells were assessed by flow cytometry. Luciferase-expressing U251 glioma cells, known to harbor a CSC niche, were intracranially implanted in immunocompromised mice. Mice were treated with DAC before the intracerebroventricular delivery of TCR-T or control T cells. Tumor growth was tracked by bioluminescent imaging (BLI), and organ tissues were harvested for analysis at the survival endpoint. RESULTS: Our findings indicate that DAC exposure significantly upregulates NY-ESO-1 messenger RNA and protein expression, alongside increases in MHC class, bolstering the immune response against patient-derived CSCs. DAC significantly sensitizes CSC 38b cells to lysis by TCR-T cells, exhibiting a marked increase in killing efficacy compared to control T cells (36% versus 20%, p=0.02) within 20 hours. In vivo, TCR-T cells delayed tumor progression and extended survival in glioma-bearing mice compared to the control cohorts (median 40 versus 27 days, p = 0.031). Furthermore, TCR-T cells successfully infiltrated tumors with preferential targeting of NY-ESO-1-expressing cancer cells and CD133-positive stem-like cells within the intricate tumor microenvironment. CONCLUSIONS: Our findings demonstrate the potential of a novel NY-ESO-1-specific TCR-T cell as a promising immunotherapeutic strategy against GBM and its difficult-to-treat CSC population. These findings underscore the need to develop TCR-based therapies further for more advanced, precision immunotherapies.