TO WILMS TUMOR PROTEIN ( WT 1 ) IN MYELODYSPLASIA RESPONSIVE TO IMMUNOSUPPRESSIVE THERAPY ( IST )

Clinical observations and laboratory evidence link bone marrow failure in myelodysplastic syndrome (MDS) to a T cell-mediated immune process that is responsive to immunosuppressive treatment (IST) in some patients. We previously reported that trisomy 8 MDS was more likely to respond to IST and was associated with skewed T cell receptor V profiles with clonally expanded CD8+ T cells capable of suppressing the growth of aneuploid progenitor cells in vitro. Furthermore, microarray analyses showed that Wilms tumor protein (WT1) was over-expressed by trisomy 8 hematopoietic progenitor (CD34+) cells as compared to CD34+ cells from healthy donors. Here we show that WT1 mRNA expression is upregulated by as much as 1,000-fold in the bone marrow mononuclear cells (BMMNC) of MDS patients with trisomy 8 (p = 0.001); WT1 protein levels were also significantly elevated. In addition, using a combination of physical and functional assays to detect the presence and biological reactivity of specific T cells respectively, we demonstrate that IST-responsive MDS patients exhibit significant CD4+ and CD8+ T cell responses directed against WT1. Finally WT1-specific CD8+ T cells are present within expanded V subfamilies and can inhibit hematopoiesis when added to autologous patient bone marrow cells in culture. Thus, our results strongly implicate WT1 as one of the antigens that triggers T cell-mediated myelosuppression in MDS. 2 N at ur e P re ce di ng s : h dl :1 01 01 /n pr e. 20 10 .4 15 4. 1 : P os te d 12 J an 2 01 0

[1]  E. Thiel,et al.  A clinical and immunologic phase 2 trial of Wilms tumor gene product 1 (WT1) peptide vaccination in patients with AML and MDS. , 2009, Blood.

[2]  J. Karbach,et al.  WT1 peptide-specific T cells generated from peripheral blood of healthy donors: possible implications for adoptive immunotherapy after allogeneic stem cell transplantation , 2009, Leukemia.

[3]  D. Price,et al.  Tricks with tetramers: how to get the most from multimeric peptide–MHC , 2009, Immunology.

[4]  John Sidney,et al.  Cross-presentation of caspase-cleaved apoptotic self antigens in HIV infection , 2007, Nature Medicine.

[5]  K. Rezvani,et al.  Leukemia-associated antigen-specific T-cell responses following combined PR1 and WT1 peptide vaccination in patients with myeloid malignancies. , 2007, Blood.

[6]  D. Douek,et al.  Graft-versus-leukemia effects associated with detectable Wilms tumor-1 specific T lymphocytes after allogeneic stem-cell transplantation for acute lymphoblastic leukemia. , 2007, Blood.

[7]  G. Mufti,et al.  CD4+CD25high Foxp3+ regulatory T cells in myelodysplastic syndrome (MDS). , 2007, Blood.

[8]  D. Scheinberg,et al.  Peptide Epitopes from the Wilms' Tumor 1 Oncoprotein Stimulate CD4+ and CD8+ T Cells That Recognize and Kill Human Malignant Mesothelioma Tumor Cells , 2007, Clinical Cancer Research.

[9]  K. Kok,et al.  High-resolution mapping identifies a commonly amplified 11q13.3 region containing multiple genes flanked by segmental duplications , 2007, Human Genetics.

[10]  E. Thiel,et al.  Identification of a Highly Immunogenic HLA-A*01-Binding T Cell Epitope of WT1 , 2006, Clinical Cancer Research.

[11]  N. Young,et al.  CD34 cells from patients with trisomy 8 myelodysplastic syndrome (MDS) express early apoptotic markers but avoid programmed cell death by up-regulation of antiapoptotic proteins. , 2006, Blood.

[12]  Allen R. Chen,et al.  The antiapoptotic gene A1/BFL1 is a WT1 target gene that mediates granulocytic differentiation and resistance to chemotherapy. , 2006, Blood.

[13]  K. Keyvanfar,et al.  T-bet, a Th1 transcription factor, is up-regulated in T cells from patients with aplastic anemia. , 2006, Blood.

[14]  D. Douek,et al.  T-Cell Responses Directed against Multiple HLA-A*0201-Restricted Epitopes Derived from Wilms' Tumor 1 Protein in Patients with Leukemia and Healthy Donors: Identification, Quantification, and Characterization , 2005, Clinical Cancer Research.

[15]  S. Kajigaya,et al.  Distinctive gene expression profiles of CD34 cells from patients with myelodysplastic syndrome characterized by specific chromosomal abnormalities. , 2004, Blood.

[16]  T. Kyo,et al.  Induction of WT1 (Wilms' tumor gene)-specific cytotoxic T lymphocytes by WT1 peptide vaccine and the resultant cancer regression. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[17]  F. Watzinger,et al.  Evaluation of candidate control genes for diagnosis and residual disease detection in leukemic patients using ‘real-time’ quantitative reverse-transcriptase polymerase chain reaction (RQ-PCR) – a Europe against cancer program , 2003, Leukemia.

[18]  D. Douek,et al.  Functional leukemia-associated antigen-specific memory CD8+ T cells exist in healthy individuals and in patients with chronic myelogenous leukemia before and after stem cell transplantation. , 2003, Blood.

[19]  K. Keyvanfar,et al.  Cytogenetic abnormalities in paroxysmal nocturnal haemoglobinuria usually occur in haematopoietic cells that are glycosylphosphatidylinositol‐anchored protein (GPI‐AP) positive , 2003, British journal of haematology.

[20]  N. Young,et al.  Preferential suppression of trisomy 8 compared with normal hematopoietic cell growth by autologous lymphocytes in patients with trisomy 8 myelodysplastic syndrome. , 2003, Blood.

[21]  I. Kawase,et al.  Wilms tumor gene peptide-based immunotherapy for patients with overt Leukemia from myelodysplastic syndrome (MDS) or MDS with myelofibrosis , 2003, International journal of hematology.

[22]  A. Levis,et al.  Significant correlation between the degree of WT1 expression and the International Prognostic Scoring System Score in patients with myelodysplastic syndromes. , 2003, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[23]  J. Goldman,et al.  Human cytotoxic T lymphocytes specific for Wilms’ tumor antigen-1 inhibit engraftment of leukemia-initiating stem cells in non-obese diabetic-severe combined immunodeficient recipients , 2003, Transplantation.

[24]  J. Crolla,et al.  Frequent chromosome aberrations revealed by molecular cytogenetic studies in patients with aniridia. , 2002, American journal of human genetics.

[25]  M. Radic,et al.  Blebs and Apoptotic Bodies Are B Cell Autoantigens1 , 2002, The Journal of Immunology.

[26]  N. Young,et al.  Distinct clinical outcomes for cytogenetic abnormalities evolving from aplastic anemia. , 2002, Blood.

[27]  N. Hosen,et al.  patients with hematopoietic malignancies product in WT 1 Humoral immune responses against Wilms tumor gene , 2002 .

[28]  N. Geller,et al.  A pilot study of the recombinant soluble human tumour necrosis factor receptor (p75)‐Fc fusion protein in patients with myelodysplastic syndrome , 2002, British journal of haematology.

[29]  A. Tsuboi,et al.  WT1 as a novel target antigen for cancer immunotherapy. , 2002, Current cancer drug targets.

[30]  L. Carter,et al.  WT1-specific serum antibodies in patients with leukemia. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[31]  J. Goldman,et al.  Selective elimination of leukemic CD34(+) progenitor cells by cytotoxic T lymphocytes specific for WT1. , 2000, Blood.

[32]  H. Sugiyama,et al.  Human cytotoxic T-lymphocyte responses specific for peptides of the wild-type Wilms' tumor gene (WT1 ) product , 2000, Immunogenetics.

[33]  Y. Oji,et al.  Constitutive expression of the Wilms' tumor gene WT1 inhibits the differentiation of myeloid progenitor cells but promotes their proliferation in response to granulocyte-colony stimulating factor (G-CSF). , 1999, Leukemia research.

[34]  Y. Oji,et al.  Wilms' tumor gene (WT1) competes with differentiation-inducing signal in hematopoietic progenitor cells. , 1998, Blood.

[35]  M A Nowak,et al.  Quantitation of HIV-1-specific cytotoxic T lymphocytes and plasma load of viral RNA. , 1998, Science.

[36]  J. York,et al.  Development of a candidate HLA A*0201 restricted peptide-based vaccine against human cytomegalovirus infection. , 1997, Blood.

[37]  N. Young,et al.  Expression and modulation of cellular receptors for interferon‐γ, tumour necrosis factor, and Fas on human bone marrow CD34+ cells , 1997, British journal of haematology.

[38]  N. Young,et al.  Fas antigen expression on CD34+ human marrow cells is induced by interferon gamma and tumor necrosis factor alpha and potentiates cytokine-mediated hematopoietic suppression in vitro. , 1995, Blood.

[39]  H. Gralnick,et al.  Proposals for the classification of the myelodysplastic syndromes , 1982, British journal of haematology.

[40]  D. Douek,et al.  preferentially reside in the bone marrow , 2008 .

[41]  K. Rezvani,et al.  The role of the immune system in myelodysplasia: implications for therapy. , 2008, Seminars in hematology.

[42]  J. Melo,et al.  Molecular profiling of CD34+ cells identifies low expression of CD7, along with high expression of proteinase 3 or elastase, as predictors of longer survival in patients with CML. , 2006, Blood.

[43]  E. Thiel,et al.  Complete remission in a patient with recurrent acute myeloid leukemia induced by vaccination with WT1 peptide in the absence of hematological or renal toxicity , 2004, Leukemia.

[44]  S. Fujita,et al.  HLA class I-restricted lysis of leukemia cells by a CD8(+) cytotoxic T-lymphocyte clone specific for WT1 peptide. , 2000, Blood.