MHC-restricted phosphopeptides from insulin receptor substrate-2 and CDC25b offer broad-based immunotherapeutic agents for cancer.

Cancer cells display novel phosphopeptides in association with MHC class I and II molecules. In this study, we evaluated two HLA-A2-restricted phosphopeptides derived from the insulin receptor substrate (IRS)-2 and the cell-cycle regulator CDC25b. These proteins are both broadly expressed in multiple malignancies and linked to cancer cell survival. Two phosphopeptides, termed pIRS-21097-1105 and pCDC25b38-46, served as targets of strong and specific CD8 T-cell memory responses in normal human donors. We cloned T-cell receptor (TCR) cDNAs from murine CD8 T-cell lines specific for either pIRS-21097-1105 or pCDC25b38-46. Expression of these TCRs in human CD8 T cells imparted high-avidity phosphopeptide-specific recognition and cytotoxic and cytokine-secreting effector activities. Using these cells, we found that endogenously processed pIRS-21097-1105 was presented on HLA-A2(+) melanomas and breast, ovarian, and colorectal carcinomas. Presentation was correlated with the level of the Ser(1100)-phosphorylated IRS-2 protein in metastatic melanoma tissues. The highest expression of this protein was evident on dividing malignant cells. Presentation of endogenously processed pCDC25b38-46 was narrower, but still evident on HLA-A2(+) melanoma, breast carcinoma, and lymphoblastoid cells. Notably, pIRS-21097-1105-specific and pCDC25b38-46-specific TCR-expressing human CD8 T cells markedly slowed tumor outgrowth in vivo. Our results define two new antigens that may be developed as immunotherapeutic agents for a broad range of HLA-A2(+) cancers.

[1]  Jennifer G. Abelin,et al.  MHC Class I–Associated Phosphopeptides Are the Targets of Memory-like Immunity in Leukemia , 2013, Science Translational Medicine.

[2]  S. Rosenberg,et al.  T-cell receptor affinity and avidity defines antitumor response and autoimmunity in T-cell immunotherapy , 2013, Proceedings of the National Academy of Sciences.

[3]  S. Rosenberg,et al.  Treating cancer with genetically engineered T cells. , 2011, Trends in biotechnology.

[4]  M. Ross,et al.  Effect of Granulocyte/Macrophage Colony-Stimulating Factor on Circulating CD8+ and CD4+ T-Cell Responses to a Multipeptide Melanoma Vaccine: Outcome of a Multicenter Randomized Trial , 2009, Clinical Cancer Research.

[5]  P. Schellhammer,et al.  Integrated data from 2 randomized, double‐blind, placebo‐controlled, phase 3 trials of active cellular immunotherapy with sipuleucel‐T in advanced prostate cancer , 2009, Cancer.

[6]  Jie Qian,et al.  Identification of tumor-associated, MHC class II-restricted phosphopeptides as targets for immunotherapy , 2009, Proceedings of the National Academy of Sciences.

[7]  S. Rosenberg,et al.  Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. , 2009, Blood.

[8]  N. Sato,et al.  The functioning antigens: beyond just as the immunological targets , 2009, Cancer science.

[9]  D. Ray,et al.  In vivo roles of CDC25 phosphatases: biological insight into the anti-cancer therapeutic targets. , 2008, Anti-cancer agents in medicinal chemistry.

[10]  Adrian V. Lee,et al.  Insulin Receptor Substrates (IRSs) and Breast Tumorigenesis , 2008, Journal of Mammary Gland Biology and Neoplasia.

[11]  S. Elledge,et al.  A quantitative atlas of mitotic phosphorylation , 2008, Proceedings of the National Academy of Sciences.

[12]  Marie-Paule Lefranc,et al.  IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis , 2008, Nucleic Acids Res..

[13]  S. Rosenberg,et al.  Adoptive cell transfer: a clinical path to effective cancer immunotherapy , 2008, Nature Reviews Cancer.

[14]  D. Kranz,et al.  Equilibrium between host and cancer caused by effector T cells killing tumor stroma. , 2008, Cancer research.

[15]  R. Darnell,et al.  A T cell receptor associated with naturally occurring human tumor immunity , 2007, Proceedings of the National Academy of Sciences.

[16]  T. Schumacher,et al.  Selecting highly affine and well-expressed TCRs for gene therapy of melanoma. , 2007, Blood.

[17]  A. Little,et al.  Monoclonal T-cell receptors: new reagents for cancer therapy. , 2007, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  Adrian V. Lee,et al.  Oncogenic Transformation by the Signaling Adaptor Proteins Insulin Receptor Substrate (IRS)-1 and IRS-2 , 2007, Cell cycle.

[19]  R. Weichselbaum,et al.  Induced sensitization of tumor stroma leads to eradication of established cancer by T cells , 2007, The Journal of experimental medicine.

[20]  R. Effros.  Role of T lymphocyte replicative senescence in vaccine efficacy. , 2007, Vaccine.

[21]  S. Rosenberg,et al.  Gene Transfer of Tumor-Reactive TCR Confers Both High Avidity and Tumor Reactivity to Nonreactive Peripheral Blood Mononuclear Cells and Tumor-Infiltrating Lymphocytes1 , 2006, The Journal of Immunology.

[22]  G. Parmigiani,et al.  The Consensus Coding Sequences of Human Breast and Colorectal Cancers , 2006, Science.

[23]  Adrian V. Lee,et al.  Mammary Tumorigenesis and Metastasis Caused by Overexpression of Insulin Receptor Substrate 1 (IRS-1) or IRS-2 , 2006, Molecular and Cellular Biology.

[24]  S. Rosenberg,et al.  Cancer Regression in Patients After Transfer of Genetically Engineered Lymphocytes , 2006, Science.

[25]  Anne M Evans,et al.  Identification of class I MHC-associated phosphopeptides as targets for cancer immunotherapy , 2006, Proceedings of the National Academy of Sciences.

[26]  J. Rossjohn,et al.  Tumors reveal their secrets to cytotoxic T cells , 2006, Proceedings of the National Academy of Sciences.

[27]  Yangbing Zhao,et al.  Enhanced antitumor activity of murine-human hybrid T-cell receptor (TCR) in human lymphocytes is associated with improved pairing and TCR/CD3 stability. , 2006, Cancer research.

[28]  C. Desbois-Mouthon,et al.  Overexpression of insulin receptor substrate-2 in human and murine hepatocellular carcinoma. , 2005, The American journal of pathology.

[29]  J. Ptak,et al.  Colorectal cancer: Mutations in a signalling pathway , 2005, Nature.

[30]  Lloyd J. Old,et al.  Cancer/testis antigens, gametogenesis and cancer , 2005, Nature Reviews Cancer.

[31]  E. Bissonette,et al.  Immunologic and clinical outcomes of vaccination with a multiepitope melanoma peptide vaccine plus low-dose interleukin-2 administered either concurrently or on a delayed schedule. , 2004, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[32]  L. Shaw,et al.  Involvement of Insulin Receptor Substrate 2 in Mammary Tumor Metastasis , 2004, Molecular and Cellular Biology.

[33]  A. Lindqvist,et al.  Characterisation of Cdc25B localisation and nuclear export during the cell cycle and in response to stress , 2004, Journal of Cell Science.

[34]  S. Rosenberg,et al.  Cancer immunotherapy: moving beyond current vaccines , 2004, Nature Medicine.

[35]  R. Schreiber,et al.  The three Es of cancer immunoediting. , 2004, Annual review of immunology.

[36]  E. Feldman,et al.  Insulin-like growth factor-I signaling in human neuroblastoma cells , 2004, Oncogene.

[37]  G. Reifenberger,et al.  Genetic Alterations and Aberrant Expression of Genes Related to the Phosphatidyl‐lnositol‐3′‐Kinase/Protein Kinase B (Akt) Signal Transduction Pathway in Glioblastomas , 2003, Brain pathology.

[38]  Yao-Tseng Chen,et al.  Cancer/testis antigens: an expanding family of targets for cancer immunotherapy , 2002, Immunological reviews.

[39]  D. Yee,et al.  Regulation of breast cancer cell motility by insulin receptor substrate-2 (IRS-2) in metastatic variants of human breast cancer cell lines , 2001, Oncogene.

[40]  Forest M. White,et al.  Phosphorylated Peptides Are Naturally Processed and Presented by Major Histocompatibility Complex Class I Molecules in Vivo , 2000, The Journal of experimental medicine.

[41]  T. Hunter,et al.  Signaling—2000 and Beyond , 2000, Cell.

[42]  D. Speiser,et al.  High Frequencies of Naive Melan-a/Mart-1–Specific Cd8+ T Cells in a Large Proportion of Human Histocompatibility Leukocyte Antigen (Hla)-A2 Individuals , 1999, The Journal of experimental medicine.

[43]  J. Shabanowitz,et al.  The peptide recognized by HLA-A68.2-restricted, squamous cell carcinoma of the lung-specific cytotoxic T lymphocytes is derived from a mutated elongation factor 2 gene. , 1998, Cancer research.

[44]  M. Korc,et al.  Enhanced expression of the insulin receptor substrate-2 docking protein in human pancreatic cancer. , 1998, Cancer research.

[45]  T. Eberlein.  gp100 Peptide Vaccine and Interleukin-2 in Patients with Advanced Melanoma , 2012 .

[46]  S. Rosenberg,et al.  High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation. , 2006, Molecular therapy : the journal of the American Society of Gene Therapy.

[47]  A. Sette,et al.  In vitro immunization and expansion of antigen-specific cytotoxic T lymphocytes for adoptive immunotherapy using peptide-pulsed dendritic cells. , 1998, Critical reviews in immunology.