Lack of specific gamma-retroviral vector long terminal repeat promoter silencing in patients receiving genetically engineered lymphocytes and activation upon lymphocyte restimulation.

Retroviral transduction of tumor antigen-specific T-cell receptor (TCR) genes into lymphocytes redirects T cells to lyse tumors. Furthermore, adoptive transfer of these lymphocytes has mediated objective responses in patients with metastatic cancer. From 2004 to 2006, more than 40 patients were treated with autologous gene-modified lymphocytes expressing a melanoma antigen-specific TCR at the National Cancer Institute. Eighteen such patients were analyzed for persistence and gene expression in vivo. In addition, the impact of epigenetic silencing and of lymphocyte restimulation was studied. Although gene-modified lymphocytes persisted in vivo, the shutdown of TCR transgene expression was observed. Bisulfite sequencing analysis and ex vivo DNA methyltransferase inhibition demonstrated that the decrease in gene expression did not result from DNA methylation. Surprisingly, down-regulation of vector-driven transgene transcriptional activity was not vector specific but mimicked that of endogenous genes. The decrease in TCR transgene expression, however, was reversed upon lymphocyte stimulation. These data demonstrate a lack of gamma-retroviral promoter-specific gene silencing in adoptively transferred human lymphocytes and support that transgene expression is largely affected by global cellular mechanisms. The use of immunomodulatory adjuvants, eg, vaccination or cytokine therapy, for in vivo T-cell activation may help overcome this metabolic quiescence and thus augment cellular immunotherapy-based cancer therapy.

[1]  Hao Liu,et al.  Virus-specific T cells engineered to coexpress tumor-specific receptors: persistence and antitumor activity in individuals with neuroblastoma , 2008, Nature Medicine.

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

[3]  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.

[4]  C. Bordignon,et al.  Antitumor effects of HSV-TK-engineered donor lymphocytes after allogeneic stem-cell transplantation. , 2007, Blood.

[5]  Masato Kubo,et al.  SOCS proteins, cytokine signalling and immune regulation , 2007, Nature Reviews Immunology.

[6]  C. June,et al.  Adoptive T cell therapy for cancer in the clinic. , 2007, The Journal of clinical investigation.

[7]  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.

[8]  Gang Wang,et al.  A Phase I Study on Adoptive Immunotherapy Using Gene-Modified T Cells for Ovarian Cancer , 2006, Clinical Cancer Research.

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

[10]  S. Sleijfer,et al.  Treatment of metastatic renal cell carcinoma with autologous T-lymphocytes genetically retargeted against carbonic anhydrase IX: first clinical experience. , 2006, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[11]  C. Lamers,et al.  Parallel detection of transduced T lymphocytes after immunogene therapy of renal cell cancer by flow cytometry and real-time polymerase chain reaction: implications for loss of transgene expression. , 2005, Human gene therapy.

[12]  H Clifford Lane,et al.  Preferential survival of CD4+ T lymphocytes engineered with anti-human immunodeficiency virus (HIV) genes in HIV-infected individuals. , 2005, Human gene therapy.

[13]  J. Ellis,et al.  Retrovirus silencing and vector design: relevance to normal and cancer stem cells? , 2005, Current gene therapy.

[14]  S. Rosenberg,et al.  Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions. , 2005, Human gene therapy.

[15]  S. Rosenberg,et al.  Adoptive cell transfer therapy following non-myeloablative but lymphodepleting chemotherapy for the treatment of patients with refractory metastatic melanoma. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  I. Plavec,et al.  Enhanced transgene expression in quiescent and activated human CD8+ T cells. , 2004, Human gene therapy.

[17]  P. Pasceri,et al.  Retrovirus silencing, variegation, extinction, and memory are controlled by a dynamic interplay of multiple epigenetic modifications. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  M. Brenner,et al.  Genetic modification of T lymphocytes for adoptive immunotherapy. , 2004, Molecular therapy : the journal of the American Society of Gene Therapy.

[19]  C. S. Swindle,et al.  Mutation of CpGs in the Murine Stem Cell Virus Retroviral Vector Long Terminal Repeat Represses Silencing in Embryonic Stem Cells* , 2004, Journal of Biological Chemistry.

[20]  Masato Kubo,et al.  Suppressors of cytokine signaling and immunity , 2003, Nature Immunology.

[21]  S. Rosenberg,et al.  High Efficiency TCR Gene Transfer into Primary Human Lymphocytes Affords Avid Recognition of Melanoma Tumor Antigen Glycoprotein 100 and Does Not Alter the Recognition of Autologous Melanoma Antigens , 2003, The Journal of Immunology.

[22]  S. Rosenberg,et al.  Recombinant fowlpox viruses encoding the anchor-modified gp100 melanoma antigen can generate antitumor immune responses in patients with metastatic melanoma. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[23]  S. Rosenberg,et al.  Immunization of Patients with Metastatic Melanoma Using Both Class I- and Class II-Restricted Peptides from Melanoma-Associated Antigens , 2003, Journal of immunotherapy.

[24]  Michel Sadelain,et al.  Targeting tumours with genetically enhanced T lymphocytes , 2003, Nature Reviews Cancer.

[25]  M. Raffeld,et al.  Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes , 2002, Science.

[26]  T. Schumacher,et al.  T-cell-receptor gene therapy , 2002, Nature Reviews Immunology.

[27]  C. S. Swindle,et al.  Mechanisms that regulate silencing of gene expression from retroviral vectors. , 2002, Journal of hematotherapy & stem cell research.

[28]  L. Notarangelo,et al.  Immune reconstitution in ADA-SCID after PBL gene therapy and discontinuation of enzyme replacement , 2002, Nature Medicine.

[29]  M. Groudine,et al.  Methylation-Mediated Proviral Silencing Is Associated with MeCP2 Recruitment and Localized Histone H3 Deacetylation , 2001, Molecular and Cellular Biology.

[30]  James R. Smith,et al.  DNA Methyltransferase Inhibition in Normal Human Fibroblasts Induces a p21-dependent Cell Cycle Withdrawal* , 2001, The Journal of Biological Chemistry.

[31]  Cameron S. Osborne,et al.  Retrovirus vector silencing is de novo methylase independent and marked by a repressive histone code , 2000, The EMBO journal.

[32]  M. Szyf,et al.  DNA Methyltransferase Inhibition Induces the Transcription of the Tumor Suppressor p21 WAF1/CIP1/sdi1 * , 2000, The Journal of Biological Chemistry.

[33]  M. van Glabbeke,et al.  New guidelines to evaluate the response to treatment in solid tumors , 2000, Journal of the National Cancer Institute.

[34]  M Van Glabbeke,et al.  New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. , 2000, Journal of the National Cancer Institute.

[35]  D. Williams,et al.  Costimulation of transduced T lymphocytes via T cell receptor-CD3 complex and CD28 leads to increased transcription of integrated retrovirus. , 1999, Human gene therapy.

[36]  D. Kohn,et al.  High-resolution analysis of cytosine methylation in the 5ĺong terminal repeat of retroviral vectors. , 1998, Human gene therapy.

[37]  L. Lum,et al.  T cell activation modulates retrovirus-mediated gene expression. , 1998, Human gene therapy.

[38]  David A. Williams,et al.  High-Efficiency Gene Transfer into Normal and Adenosine Deaminase-Deficient T Lymphocytes Is Mediated by Transduction on Recombinant Fibronectin Fragments , 1998, Journal of Virology.

[39]  F. Marincola,et al.  Immunologic and therapeutic evaluation of a synthetic peptide vaccine for the treatment of patients with metastatic melanoma , 1998, Nature Medicine.

[40]  P. Romero,et al.  Enhanced generation of specific tumor-reactive CTL in vitro by selected Melan-A/MART-1 immunodominant peptide analogues. , 1998, Journal of immunology.

[41]  G. Nabel,et al.  Enhanced T cell engraftment after retroviral delivery of an antiviral gene in HIV-infected individuals. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[42]  R. Morgan,et al.  Efficient in vivo marking of primary CD4+ T lymphocytes in nonhuman primates using a gibbon ape leukemia virus-derived retroviral vector. , 1997, Blood.

[43]  W. Anderson,et al.  Molecular analysis of T lymphocyte-directed gene therapy for adenosine deaminase deficiency: long-term expression in vivo of genes introduced with a retroviral vector. , 1996, Human Gene Therapy.

[44]  D. Kohn,et al.  Expression levels by retroviral vectors based upon the N2 and the MFG backbones. , 1996, Gene therapy.

[45]  R. Mulligan,et al.  Increased levels of spliced RNA account for augmented expression from the MFG retroviral vector in hematopoietic cells. , 1996, Gene therapy.

[46]  S. Rosenberg,et al.  T Lymphocyte-Directed Gene Therapy for ADA− SCID: Initial Trial Results After 4 Years , 1995, Science.

[47]  Evelina Mazzolari,et al.  Gene Therapy in Peripheral Blood Lymphocytes and Bone Marrow for ADA− Immunodeficient Patients , 1995, Science.

[48]  T. Hawley,et al.  Versatile retroviral vectors for potential use in gene therapy. , 1994, Gene therapy.

[49]  S. Riddell,et al.  The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells. , 1990, Journal of immunological methods.

[50]  Rudolf Jaenisch,et al.  De novo methylation and expression of retroviral genomes during mouse embryogenesis , 1982, Nature.

[51]  R. Jaenisch,et al.  De novo methylation, expression, and infectivity of retroviral genomes introduced into embryonal carcinoma cells. , 1982, Proceedings of the National Academy of Sciences of the United States of America.