Dietary administration of alpha-tocopheryloxyacetic acid induces CD4+ T cell activation and prolongs survival in murine rhabdomyosarcoma

Relapsed sarcomas remain essentially incurable for most children with cancer. Alpha-Tocopheryloxyacetic acid (αTEA) is a scalable semi-synthetic derivative of alpha-tocopherol that has shown promising activity preclinically and clinically in adult cancer, but has not yet been studied in pediatric sarcoma. We hypothesized that αTEA would mediate anti-tumor effects in rhabdomyosarcoma (RMS) by modulating the immunosuppressive milieu. Treatment of murine M3-9-M RMS in vitro with αTEA (2.5–100uM) resulted in increased apoptosis by flow cytometry (Annexin V+/7AAD+) and Incucyte analysis (Annexin V+) within 24 hours (p < 0.05). Using an established orthotopic luciferase+ M3-9-M in vivo RMS model, αTEA-supplemented chow administered for 21 days significantly reduced tumor volume (p < 0.001) and bioluminescence (p < 0.05) by 28 days, resulting in a significant prolongation of survival compared to matched control chow (p < 0.05), up to 4 weeks after the feed was stopped. Analysis of spleens from RMS mice fed αTEA-supplemented diets showed an increased percentage of Ki-67+ and IFNγ+CD4+ T cells (p < 0.01) at the end of treatment as compared to recipients of the control diet. In addition, a decrease in the percentage of CD11b+Arg-1+ (p < 0.01) and PD-L1+ (p < 0.05) cells within the spleen were observed in αTEA-treated RMS tumor-bearing mice. RNA-Seq analysis of RMS tumors is underway to explore changes in αTEA vs. control-treated mice. These results suggest that αTEA causes direct induction of apoptosis of RMS in vitro and in vivo, leading to prolonged survival through enhanced CD4+ T cell stimulation and reduced myeloid cell-mediated immunosuppression in RMS-bearing mice. Further investigation of αTEA as an immunomodulating agent is warranted.