Abstract 1584: MHC-restricted phosphopeptides as broad-based immunotherapeutic targets for cancer
Authors: Zarling Angela L. , Obeng Rebecca C. , Cummings Kara L. , Desch Ashley N. , English Ann M. , Cottine Jennifer M. , Norris Andrew , Brill Louis B. , Deacon Donna H. , Conaway Mark , Shabanowitz Jeffrey , Hunt Donald F. , Slingluff Craig L. , Engelhard Victor H. 
Published: August 2012

Identification of the antigens (Ags) recognized by CD8+ T cells specific for melanoma and other types of cancer cells has been motivated by the expectation that they will serve as the basis for a therapeutic vaccine or will be useful for the stimulation of more effective CD8+ CTL for adoptive immunotherapy. Few of the currently identified Ags are derived from source proteins directly associated with transformation or metastasis, and many are self-Ags which may be subject to tolerance mechanisms. We have defined a new array of phosphorylated peptides presented by HLA-A1, -A2, -A3, -B7, -B44 and HLA-B27 that are presented on melanomas from multiple patients at &gt;1 copy per cell and derived from source proteins associated with tumorigenesis and metastasis. We evaluated the immunogenicity of several of the HLA-A2-restricted phosphopeptide Ags in an HLA-transgenic murine model system and in vitro human T cell cultures, evaluated the presence of the phosphorylated source protein or display of the phosphorylated epitope on cancers of different histological types, and investigated the ability of phosphopeptide-specific CD8+ T cells to prevent outgrowth of cancer cells in a xenogenic model. Evaluation of metastatic melanoma tissue microarrays and cancer cell lysates demonstrated that the phosphorylated source proteins for β-catenin (Ser33-phosphorylated), Breast cancer anti-estrogen resistance −3 (BCAR3; Thr130-phosphorylated), and Insulin receptor substrate-2 (IRS2; Ser1100-phosphorylated) were broadly expressed in melanomas and other cancers but not normal tissues. In addition, these epitopes can be utilized to elicit phosphopeptide-specific T cells in both murine HLA-A2 transgenic mice in vivo and human in vitro culture. The phospho-β-catenin, phospho-IRS2 and phospho-BCAR3-specific-T cells that were generated differentiate between the phosphorylated and non-modified forms of their respective peptides and recognize the endogenously processed and presented phosphopeptide on the surface of cancer cells. Finally, adoptive transfer of these phosphopeptide-specific T cells slowed outgrowth of melanoma tumors implanted in immunodeficient NOD.SCID.IL2Rα-/- mice. Thus, we have defined a new set of post-translationally modified epitopes derived from source proteins associated with tumorigenesis and metastasis that are immunogenic and capable of slowing tumor outgrowth in a xenograft model. Based on these results, we are currently planning a clinical trial utilizing these HLA-A2-restricted phosphopeptides in melanoma patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 1584. doi:1538-7445.AM2012-1584