Home LiteratureArticle Details
PMID: 19494842 Published · ppublish English Journal Article Research Support, N.I.H., Intramural

Efficient nonviral Sleeping Beauty transposon-based TCR gene transfer to peripheral blood lymphocytes confers antigen-specific antitumor reactivity.

Gene therapy ·Vol. 16 ·No. 8 ·2009-08-00 ·Pages 1042-9

Peng PD, Cohen CJ, Yang S, Hsu C, Jones S, Zhao Y, Zheng Z, Rosenberg SA, Morgan RA

Abstract

Genetically engineered lymphocytes hold promise for the treatment of genetic disease, viral infections and cancer. However, current methods for genetic transduction of peripheral blood lymphocytes rely on viral vectors, which are hindered by production and safety-related problems. In this study, we demonstrated an efficient novel nonviral platform for gene transfer to lymphocytes. The Sleeping Beauty transposon-mediated approach allowed for long-term stable expression of transgenes at approximately 50% efficiency. Utilizing transposon constructs expressing tumor antigen-specific T-cell receptor genes targeting p53 and MART-1, we demonstrated sustained expression and functional reactivity of transposon-engineered lymphocytes on encountering target antigen presented on tumor cells. We found that transposon- and retroviral-modified lymphocytes had comparable transgene expression and phenotypic function. These results demonstrate the promise of nonviral ex vivo genetic modification of autologous lymphocytes for the treatment of cancer and immunologic disease.

MeSH Terms
Antigens, Neoplasm DNA Transposable Elements Gene Transfer Techniques Genes, T-Cell Receptor Humans Immunotherapy, Adoptive/methods T-Lymphocytes/immunology Transduction, Genetic Transposases/genetics
Chemicals
Antigens, Neoplasm DNA Transposable Elements Transposases sleeping beauty transposase, human
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Peng P D
Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Cohen C J
Yang S
Hsu C
Jones S
Zhao Y
Zheng Z
Rosenberg S A
Morgan R A
References (44)
44 references, click to expand
  1. The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells.
    J Immunol Methods. 1990 Apr 17;128(2):189-201 PMID: 1691237
  2. Targeted Sleeping Beauty transposition in human cells.
    Mol Ther. 2007 Jun;15(6):1137-44 PMID: 17426709
  3. Immunological and clinical responses in metastatic renal cancer patients vaccinated with tumor RNA-transfected dendritic cells.
    Cancer Res. 2003 May 1;63(9):2127-33 PMID: 12727829
  4. Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system.
    Nat Genet. 2000 May;25(1):35-41 PMID: 10802653
  5. Primary human lymphocytes transduced with NY-ESO-1 antigen-specific TCR genes recognize and kill diverse human tumor cell lines.
    J Immunol. 2005 Apr 1;174(7):4415-23 PMID: 15778407
  6. Sustainable correction of junctional epidermolysis bullosa via transposon-mediated nonviral gene transfer.
    Gene Ther. 2003 Jul;10(13):1099-104 PMID: 12808440
  7. Mechanisms that regulate silencing of gene expression from retroviral vectors.
    J Hematother Stem Cell Res. 2002 Jun;11(3):449-56 PMID: 12183830
  8. Stable gene transfer and expression in human primary T cells by the Sleeping Beauty transposon system.
    Blood. 2006 Jan 15;107(2):483-91 PMID: 16189271
  9. Development of hyperactive sleeping beauty transposon vectors by mutational analysis.
    Mol Ther. 2004 Feb;9(2):292-304 PMID: 14759813
  10. Redirecting specificity of T-cell populations for CD19 using the Sleeping Beauty system.
    Cancer Res. 2008 Apr 15;68(8):2961-71 PMID: 18413766
  11. Transfer of a TCR gene derived from a patient with a marked antitumor response conveys highly active T-cell effector functions.
    Hum Gene Ther. 2005 Apr;16(4):457-72 PMID: 15871677
  12. Lentivirus vector-mediated expression of tumor-associated epitopes by human antigen presenting cells.
    Hum Gene Ther. 2004 Apr;15(4):393-404 PMID: 15053864
  13. Cancer regression in patients after transfer of genetically engineered lymphocytes.
    Science. 2006 Oct 6;314(5796):126-9 PMID: 16946036
  14. A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer.
    Clin Cancer Res. 2006 Oct 15;12(20 Pt 1):6106-15 PMID: 17062687
  15. Human T lymphocytes transduced by lentiviral vectors in the absence of TCR activation maintain an intact immune competence.
    Blood. 2003 Jul 15;102(2):497-505 PMID: 12649146
  16. Acquisition of full effector function in vitro paradoxically impairs the in vivo antitumor efficacy of adoptively transferred CD8+ T cells.
    J Clin Invest. 2005 Jun;115(6):1616-26 PMID: 15931392
  17. Correction of multi-gene deficiency in vivo using a single 'self-cleaving' 2A peptide-based retroviral vector.
    Nat Biotechnol. 2004 May;22(5):589-94 PMID: 15064769
  18. Lentivirus-mediated gene transfer and expression in established human tumor antigen-specific cytotoxic T cells and primary unstimulated T cells.
    Hum Gene Ther. 2003 Jul 20;14(11):1089-105 PMID: 12885348
  19. Adoptive cell transfer therapy following non-myeloablative but lymphodepleting chemotherapy for the treatment of patients with refractory metastatic melanoma.
    J Clin Oncol. 2005 Apr 1;23(10):2346-57 PMID: 15800326
  20. Site-directed transposon integration in human cells.
    Nucleic Acids Res. 2007;35(7):e50 PMID: 17344320
  21. Adoptive cell transfer: a clinical path to effective cancer immunotherapy.
    Nat Rev Cancer. 2008 Apr;8(4):299-308 PMID: 18354418
  22. Adoptive immunotherapy for indolent non-Hodgkin lymphoma and mantle cell lymphoma using genetically modified autologous CD20-specific T cells.
    Blood. 2008 Sep 15;112(6):2261-71 PMID: 18509084
  23. 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.
    J Immunol. 2003 Sep 15;171(6):3287-95 PMID: 12960359
  24. Target-site preferences of Sleeping Beauty transposons.
    J Mol Biol. 2005 Feb 11;346(1):161-73 PMID: 15663935
  25. RNA as a source of transposase for Sleeping Beauty-mediated gene insertion and expression in somatic cells and tissues.
    Mol Ther. 2006 Mar;13(3):625-30 PMID: 16368272
  26. Adoptive cell transfer therapy.
    Semin Oncol. 2007 Dec;34(6):524-31 PMID: 18083376
  27. Supernatural T cells: genetic modification of T cells for cancer therapy.
    Nat Rev Immunol. 2005 Dec;5(12):928-40 PMID: 16322746
  28. Recognition of fresh human tumor by human peripheral blood lymphocytes transduced with a bicistronic retroviral vector encoding a murine anti-p53 TCR.
    J Immunol. 2005 Nov 1;175(9):5799-808 PMID: 16237072
  29. Enhanced antitumor activity of murine-human hybrid T-cell receptor (TCR) in human lymphocytes is associated with improved pairing and TCR/CD3 stability.
    Cancer Res. 2006 Sep 1;66(17):8878-86 PMID: 16951205
  30. A serious adverse event after successful gene therapy for X-linked severe combined immunodeficiency.
    N Engl J Med. 2003 Jan 16;348(3):255-6 PMID: 12529469
  31. High-resolution genome-wide mapping of transposon integration in mammals.
    Mol Cell Biol. 2005 Mar;25(6):2085-94 PMID: 15743807
  32. High-efficiency transfection of primary human and mouse T lymphocytes using RNA electroporation.
    Mol Ther. 2006 Jan;13(1):151-9 PMID: 16140584
  33. Silencing of gene expression: implications for design of retrovirus vectors.
    Rev Med Virol. 2001 Jul-Aug;11(4):205-17 PMID: 11479927
  34. Adoptive transfer of chimeric antigen receptor re-directed cytolytic T lymphocyte clones in patients with neuroblastoma.
    Mol Ther. 2007 Apr;15(4):825-33 PMID: 17299405
  35. Gene transfer of tumor-reactive TCR confers both high avidity and tumor reactivity to nonreactive peripheral blood mononuclear cells and tumor-infiltrating lymphocytes.
    J Immunol. 2006 Nov 1;177(9):6548-59 PMID: 17056587
  36. Sustained FVIII expression and phenotypic correction of hemophilia A in neonatal mice using an endothelial-targeted sleeping beauty transposon.
    Mol Ther. 2006 May;13(5):1006-15 PMID: 16464640
  37. Transcription start regions in the human genome are favored targets for MLV integration.
    Science. 2003 Jun 13;300(5626):1749-51 PMID: 12805549
  38. Treatment of metastatic renal cell carcinoma with autologous T-lymphocytes genetically retargeted against carbonic anhydrase IX: first clinical experience.
    J Clin Oncol. 2006 May 1;24(13):e20-2 PMID: 16648493
  39. Mutational analysis of the N-terminal DNA-binding domain of sleeping beauty transposase: critical residues for DNA binding and hyperactivity in mammalian cells.
    Mol Cell Biol. 2004 Oct;24(20):9239-47 PMID: 15456893
  40. Sleeping beauty transposon-mediated gene therapy for prolonged expression.
    Adv Genet. 2005;54:189-232 PMID: 16096013
  41. Sleeping Beauty transposon-mediated engineering of human primary T cells for therapy of CD19+ lymphoid malignancies.
    Mol Ther. 2008 Mar;16(3):580-9 PMID: 18227839
  42. Cooperation of human tumor-reactive CD4+ and CD8+ T cells after redirection of their specificity by a high-affinity p53A2.1-specific TCR.
    Immunity. 2005 Jan;22(1):117-29 PMID: 15664164
  43. Gene transfer into genomes of human cells by the sleeping beauty transposon system.
    Mol Ther. 2003 Jul;8(1):108-17 PMID: 12842434
  44. Transposons for gene therapy!
    Curr Gene Ther. 2006 Oct;6(5):593-607 PMID: 17073604
Article Info
Journal
Gene therapy
Abbr.
Gene Ther
ISSN
1476-5462
DOI
10.1038/gt.2009.54
Published
2009-08-00
Epub
2009-00-04
Pages
1042-9
Language
English
Region
England
NLM ID
9421525
PMCID
PMC3469249
Subset
IM
Grants
Intramural NIH HHS · Z01 SC003811-33 · United States
External Links
PubMed source DOI: 10.1038/gt.2009.54 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2