Ewing Sarcoma (EwS) is the second most common malignant bone tumor predominantly affecting children, adolescents, and young adults. The hallmark genetic aberration in EwS is characterized by a unique t(11:22) (q24;q12) translocation resulting in the pathognomonic EWSR1::FLI1 fusion oncoprotein. This chimeric transcription factor plays a central role in EwS pathogenesis by disrupting the normal gene regulatory network. While EwS is often considered an immunologically "cold tumor" due to its low mutational burden, emerging evidence has shed light on the dysregulation of specific target genes of EWSR1::FLI1 as potential immunotherapeutic targets. The EWSR1::FLI1 onco-fusion drives a unique 32 gene signature which includes Lipoxygenase Homology Domains 1 (LOX¬HD1), an aberrantly expressed stereociliary protein required for EwS tumorigenesis. As standard of care therapies for EwS fail to provide favorable outcomes for patients with metastatic disease, there is a need for continued research in this field. In this thesis, we assessed the potential of LOXHD1 as an immunotherapeutic target. Utilizing computational prediction algorithms coupled with immunopeptidomic analysis, we report for the first time several high affinity HLA (human leukocyte antigen) class I restricted LOXHD1 epitopes in EwS. We demonstrate that LOXHD1 immunogenic epitopes restricted to HLA-A02:01 allowed for the isolation of high avidity TCRs. LOXHD1-specific TCR engineered CD8+ T cells confer cytotoxic activity against a panel of HLA-A02:01+ EwS tumor cell lines and adoptive transfer leads to tumor eradication in a mouse xenograft model of EwS. Our findings elucidate LOXHD1 as an onco-fusion regulated / non-mutated EwS tumor associated antigen (TAA) with expression limited to inner hair cells of the cochlea, adult testis, and EwS.