Azacitidine induces demethylation of the Epstein-Barr virus genome in tumors.

PURPOSE To determine whether therapy with a DNA methyltransferase inhibitor is effective in achieving demethylation and gene re-expression in tumor DNA in patients. METHODS Biopsy specimens were obtained from patients with Epstein-Barr virus-associated tumors, enrolled on a clinical trial of 5-azacitidine, within 72 hours of the conclusion of the last infusion of the first cycle of therapy, and compared to pretreatment specimens. Methylation-specific polymerase chain reaction, bisulfite genomic sequencing, and immunohistochemistry were used to assess demethylation and gene re-expression. RESULTS Substantial degrees of demethylation were detected in all latent and lytic Epstein-Barr virus promoters examined. Immunohistochemistry suggested activation of a previously silent viral antigen expression in one instance. CONCLUSION Pharmacologic reversal of dense CpG methylation in tumor tissue can be achieved in patients.

[1]  J. Holland,et al.  Randomized controlled trial of azacitidine in patients with the myelodysplastic syndrome: a study of the cancer and leukemia group B. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[2]  Bert Vogelstein,et al.  DNMT1 and DNMT3b cooperate to silence genes in human cancer cells , 2002, Nature.

[3]  C. Plass,et al.  HLTF gene silencing in human colon cancer , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Peter A. Jones,et al.  Analysis of gene induction in human fibroblasts and bladder cancer cells exposed to the methylation inhibitor 5-aza-2'-deoxycytidine. , 2002, Cancer research.

[5]  Matty P. Weijenberg,et al.  A genomic screen for genes upregulated by demethylation and histone deacetylase inhibition in human colorectal cancer , 2002, Nature Genetics.

[6]  R. Momparler,et al.  Potential of 5-aza-2'-deoxycytidine (Decitabine) a potent inhibitor of DNA methylation for therapy of advanced non-small cell lung cancer. , 2001, Lung cancer.

[7]  R. Ambinder,et al.  Induction of Epstein-Barr Virus Kinases To Sensitize Tumor Cells to Nucleoside Analogues , 2001, Antimicrobial Agents and Chemotherapy.

[8]  A. Feinberg,et al.  Loss of imprinting of insulin-like growth factor-II in Wilms' tumor commonly involves altered methylation but not mutations of CTCF or its binding site. , 2001, Cancer research.

[9]  D. Baarle,et al.  Dysfunctional Epstein-Barr virus (EBV)-specific CD8(+) T lymphocytes and increased EBV load in HIV-1 infected individuals progressing to AIDS-related non-Hodgkin lymphoma. , 2001, Blood.

[10]  J. Herman,et al.  Promoter hypermethylation of the DNA repair gene O(6)-methylguanine-DNA methyltransferase is associated with the presence of G:C to A:T transition mutations in p53 in human colorectal tumorigenesis. , 2001, Cancer research.

[11]  J. Minárovits,et al.  Protein-DNA Binding and CpG Methylation at Nucleotide Resolution of Latency-Associated Promoters Qp, Cp, and LMP1p of Epstein-Barr Virus , 2001, Journal of Virology.

[12]  A. Feinberg,et al.  Cancer epigenetics takes center stage. , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[13]  A. Rickinson,et al.  Methylation of Transcription Factor Binding Sites in the Epstein-Barr Virus Latent Cycle Promoter Wp Coincides with Promoter Down-Regulation during Virus-Induced B-Cell Transformation , 2000, Journal of Virology.

[14]  J. Herman,et al.  Promoter-region hypermethylation and gene silencing in human cancer. , 2000, Current topics in microbiology and immunology.

[15]  I. Flinn,et al.  Application of the ELISPOT assay to the characterization of CD8(+) responses to Epstein-Barr virus antigens. , 2000, Blood.

[16]  S. Speck,et al.  Differential Methylation of Epstein-Barr Virus Latency Promoters Facilitates Viral Persistence in Healthy Seropositive Individuals , 1999, Journal of Virology.

[17]  Thierry Boon,et al.  DNA Methylation Is the Primary Silencing Mechanism for a Set of Germ Line- and Tumor-Specific Genes with a CpG-Rich Promoter , 1999, Molecular and Cellular Biology.

[18]  Q. Tao,et al.  DNA methylation and the Epstein-Barr virus. , 1999, Seminars in cancer biology.

[19]  Q. Tao,et al.  Methylation status of the Epstein-Barr virus major latent promoter C in iatrogenic B cell lymphoproliferative disease. Application of PCR-based analysis. , 1999, The American journal of pathology.

[20]  S. Rowland-Jones,et al.  A re-evaluation of the frequency of CD8+ T cells specific for EBV in healthy virus carriers. , 1999, Journal of immunology.

[21]  A. Hildesheim,et al.  The Epstein-Barr Virus Major Latent Promoter Qp Is Constitutively Active, Hypomethylated, and Methylation Sensitive , 1998, Journal of Virology.

[22]  J. Strouboulis,et al.  Methylated DNA and MeCP2 recruit histone deacetylase to repress transcription , 1998, Nature Genetics.

[23]  A. Hildesheim,et al.  Epstein-Barr virus (EBV) in endemic Burkitt's lymphoma: molecular analysis of primary tumor tissue. , 1998, Blood.

[24]  D. Srivastava,et al.  Infusion of cytotoxic T cells for the prevention and treatment of Epstein-Barr virus-induced lymphoma in allogeneic transplant recipients. , 1998, Blood.

[25]  K. Robertson,et al.  Methylation of the Epstein-Barr virus genome in normal lymphocytes. , 1997, Blood.

[26]  K. Robertson,et al.  Mapping promoter regions that are hypersensitive to methylation-mediated inhibition of transcription: application of the methylation cassette assay to the Epstein-Barr virus major latency promoter , 1997, Journal of virology.

[27]  M. Kersten,et al.  Epstein-Barr virus-specific cytotoxic T cell responses in HIV-1 infection: different kinetics in patients progressing to opportunistic infection or non-Hodgkin's lymphoma. , 1997, The Journal of clinical investigation.

[28]  M. Gulley,et al.  CpG methylation of the major Epstein-Barr virus latency promoter in Burkitt's lymphoma and Hodgkin's disease. , 1996, Blood.

[29]  Jie Yang,et al.  Epstein-Barr virus as a therapeutic target in Hodgkin's disease and nasopharyngeal carcinoma. , 1996, Seminars in cancer biology.

[30]  O. de Backer,et al.  The activation of human gene MAGE-1 in tumor cells is correlated with genome-wide demethylation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[31]  D. Samid,et al.  Transcriptional activation of the Epstein-Barr virus latency C promoter after 5-azacytidine treatment: evidence that demethylation at a single CpG site is crucial , 1995, Molecular and cellular biology.

[32]  S. Baylin,et al.  Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression. , 1995, Cancer research.

[33]  G. Srivastava,et al.  Epstein‐barr virus is localized in the tumour cells of nasal lymphomas of NK, T or B cell type , 1995, International journal of cancer.

[34]  M. Ladanyi,et al.  Infusions of donor leukocytes to treat Epstein-Barr virus-associated lymphoproliferative disorders after allogeneic bone marrow transplantation. , 1994, The New England journal of medicine.

[35]  M. van Glabbeke,et al.  The EORTC Early Clinical Trials Cooperative Group experience with 5-aza-2'-deoxycytidine (NSC 127716) in patients with colo-rectal, head and neck, renal carcinomas and malignant melanomas. , 1987, European journal of cancer & clinical oncology.

[36]  B. Leyland-Jones,et al.  Biochemistry of azacitidine: a review. , 1987, Cancer treatment reports.

[37]  H. Pinedo,et al.  Phase I and pharmacokinetic study of 5-aza-2'-deoxycytidine (NSC 127716) in cancer patients. , 1986, Cancer research.

[38]  J. Pirkle,et al.  The disposition and pharmacokinetics in humans of 5-azacytidine administered intravenously as a bolus or by continuous infusion. , 1976, Cancer research.