CoChat
Sign Up
Cited by 448CAR-T cell therapy researchCRISPR and Genetic EngineeringImmunotherapy and Immune Responses

Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer

Open access
Provasi, Elena, Genovese, Pietro, Lombardo, Angelo, Magnani, Zulma, Liu, Pei-Qi, Reik, Andreas, Chu, Victoria, Paschon, David E +13 more·2012·Nature Medicine
DOI 10.1038/nm.2700PMC PMC5019824
Open this paper in CoChat — free account

Abstract

The transfer of high-avidity T cell receptor (TCR) genes isolated from rare tumor-specific lymphocytes into polyclonal T cells is an attractive cancer immunotherapy strategy. However, TCR gene transfer results in competition for surface expression and inappropriate pairing between the exogenous and endogenous TCR chains, resulting in suboptimal activity and potentially harmful unpredicted antigen specificities of the resultant TCRs. We designed zinc-finger nucleases (ZFNs) that promoted the disruption of endogenous TCR β- and α-chain genes. Lymphocytes treated with ZFNs lacked surface expression of CD3-TCR and expanded with the addition of interleukin-7 (IL-7) and IL-15. After lentiviral transfer of a TCR specific for the Wilms tumor 1 (WT1) antigen, these TCR-edited cells expressed the new TCR at high levels, were easily expanded to near purity and were superior at specific antigen recognition compared to donor-matched, unedited TCR-transferred cells. In contrast to unedited TCR-transferred cells, the TCR-edited lymphocytes did not mediate off-target reactivity while maintaining their anti-tumor activity in vivo, thus showing that complete editing of T cell specificity generates tumor-specific lymphocytes with improved biosafety profiles.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/nm.2700

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • In a pre-clinical study, zinc finger nucleases were used in T cells to replace the endogenous TCR with a tumor-specific TCR introduced by an NILV.

    “In a pre-clinical study, zinc finger nucleases were used in T cells to replace the endogenous TCR with a tumor-specific TCR introduced by an NILV [90].”

    From Clinical use of lentiviral vectors (2018) · cites this paper as [90]

    The Introduction describes ZFN knockout of the endogenous TCR followed by transfer of a tumor-specific TCR. But the Results section on TCR β-gene editing says the TCR was delivered by integrating lentiviral vectors, and Online Methods distinguishes these from the integrase-defective vectors used to deliver the ZFNs. The claim's NILV delivery of the tumor-specific TCR is therefore incorrect. (LLM, conf=0.91)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 37

  1. [1]Appelbaum FR Haematopoietic cell transplantation as immunotherapy. Nature 2001 411 385 389 11357147
  2. [2]Dudley ME Adoptive cell therapy for patients with metastatic melanoma: evaluation of intensive myeloablative chemoradiation preparative regimens. J Clin Oncol 2008 26 5233 5239 18809613 PMC2652090
  3. [3]Hunder NN Treatment of metastatic melanoma with autologous CD4+ T cells against NY-ESO-1. N Engl J Med 2008 358 2698 2703 18565862 PMC3277288
  4. [4]Kessels HW Wolkers MC van den Boom MD van der Valk MA Schumacher TN Immunotherapy through TCR gene transfer. Nature immunology 2001 2 957 961 11577349
  5. [5]Morgan RA Cancer regression in patients after transfer of genetically engineered lymphocytes. Science 2006 314 126 129 16946036 PMC2267026
  6. [6]Johnson LA Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood 2009 114 535 546 19451549 PMC2929689
  7. [7]Robbins PF Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1. J Clin Oncol 29 917 924 21282551 PMC3068063
  8. [8]Heemskerk MH Efficiency of T-cell receptor expression in dual-specific T cells is controlled by the intrinsic qualities of the TCR chains within the TCR-CD3 complex. Blood 2007 109 235 243 16968899
  9. [9]van Loenen MM Mixed T cell receptor dimers harbor potentially harmful neoreactivity. Proceedings of the National Academy of Sciences of the United States of America 107 10972 10977 20534461 PMC2890759
  10. [10]Bendle GM Lethal graft-versus-host disease in mouse models of T cell receptor gene therapy. Nature medicine 16 565 570 561p following 570 20400962
  11. [11]Cohen CJ Zhao Y Zheng Z Rosenberg SA Morgan RA Enhanced antitumor activity of murine-human hybrid T-cell receptor (TCR) in human lymphocytes is associated with improved pairing and TCR/CD3 stability. Cancer research 2006 66 8878 8886 16951205 PMC2147082
  12. [12]Okamoto S Improved expression and reactivity of transduced tumor-specific TCRs in human lymphocytes by specific silencing of endogenous TCR. Cancer research 2009 69 9003 9011 19903853
  13. [13]Davis JL Development of human anti-murine T-cell receptor antibodies in both responding and nonresponding patients enrolled in TCR gene therapy trials. Clin Cancer Res 16 5852 5861 21138872 PMC3058233
  14. [14]Scholten KB Codon modification of T cell receptors allows enhanced functional expression in transgenic human T cells. Clinical immunology (Orlando, Fla 2006 119 135 145 16458072
  15. [15]Thomas S Targeting the Wilms tumor antigen 1 by TCR gene transfer: TCR variants improve tetramer binding but not the function of gene modified human T cells. J Immunol 2007 179 5803 5810 17947653
  16. [16]Urnov FD Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 2005 435 646 651 15806097
  17. [17]Urnov FD Rebar EJ Holmes MC Zhang HS Gregory PD Genome editing with engineered zinc finger nucleases. Nat Rev Genet 11 636 646 20717154
  18. [18]Lombardo A Gene editing in human stem cells using zinc finger nucleases and integrase-defective lentiviral vector delivery. Nature biotechnology 2007 25 1298 1306 17965707
  19. [19]Lombardo A Site-specific integration and tailoring of cassette design for sustainable gene transfer. Nature methods 8 861 869 21857672
  20. [20]Perez EE Establishment of HIV-1 resistance in CD4+ T cells by genome editing using zinc-finger nucleases. Nature biotechnology 2008 26 808 816 PMC3422503 18587387
  21. [21]Klebanoff CA Central memory self/tumor-reactive CD8+ T cells confer superior antitumor immunity compared with effector memory T cells. Proceedings of the National Academy of Sciences of the United States of America 2005 102 9571 9576 15980149 PMC1172264
  22. [22]Berger C Adoptive transfer of effector CD8+ T cells derived from central memory cells establishes persistent T cell memory in primates. J Clin Invest 2008 118 294 305 18060041 PMC2104476
  23. [23]Kaneko S IL-7 and IL-15 allow the generation of suicide gene-modified alloreactive self-renewing central memory human T lymphocytes. Blood 2009 113 1006 1015 18978209
  24. [24]Bondanza A IL-7 receptor expression identifies suicide gene-modified allospecific CD8+ T cells capable of self-renewal and differentiation into antileukemia effectors. Blood 117 6469 6478 21531977
  25. [25]Liu K Catalfamo M Li Y Henkart PA Weng NP IL-15 mimics T cell receptor crosslinking in the induction of cellular proliferation, gene expression, and cytotoxicity in CD8+ memory T cells. Proceedings of the National Academy of Sciences of the United States of America 2002 99 6192 6197 11972069 PMC122925
  26. [26]Kuball J Facilitating matched pairing and expression of TCR chains introduced into human T cells. Blood 2007 109 2331 2338 17082316 PMC1852191
  27. [27]Amendola M Venneri MA Biffi A Vigna E Naldini L Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters. Nature biotechnology 2005 23 108 116 15619618
  28. [28]Cohen CJ Enhanced antitumor activity of T cells engineered to express T-cell receptors with a second disulfide bond. Cancer research 2007 67 3898 3903 17440104 PMC2147081
  29. [29]Sollu C Autonomous zinc-finger nuclease pairs for targeted chromosomal deletion. Nucleic acids research 38 8269 8276 20716517 PMC3001086
  30. [30]Heslop HE Long-term outcome of EBV-specific T-cell infusions to prevent or treat EBV-related lymphoproliferative disease in transplant recipients. Blood 115 925 935 19880495 PMC2817637
  31. [31]Follenzi A Naldini L Generation of HIV-1 derived lentiviral vectors. Methods in enzymology 2002 346 454 465 11883085
  32. [32]Nilsson M Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells. The journal of gene medicine 2004 6 631 641 15170734
  33. [33]Doyon Y Transient cold shock enhances zinc-finger nuclease-mediated gene disruption. Nature methods 7 459 460 20436476
  34. [34]Riddell SR Greenberg PD The use of anti-CD3 and anti-CD28 monoclonal antibodies to clone and expand human antigen-specific T cells. Journal of immunological methods 1990 128 189 201 1691237
  35. [35]Sacre K Repertoire, diversity, and differentiation of specific CD8 T cells are associated with immune protection against human cytomegalovirus disease. J Exp Med 2005 201 1999 2010 15967826 PMC2212029
  36. [36]Cooke KR LPS antagonism reduces graft-versus-host disease and preserves graft-versus-leukemia activity after experimental bone marrow transplantation. J Clin Invest 2001 107 1581 1589 11413166 PMC200193
  37. [37]Bondanza A Suicide gene therapy of graft-versus-host disease induced by central memory human T lymphocytes. Blood 2006 107 1828 1836 16293601

Cited by

Sorted by citations

Authors
Provasi, E., Genovese, P., Lombardo, A., Magnani, Z., Liu, P.-Q., Reik, A., Chu, V., Paschon, D. E., Zhang, L., Kuball, J., Camisa, B., Bondanza, A., Casorati, G., Ponzoni, M., Ciceri, F., Bordignon, C., Greenberg, P. D., Holmes, M. C., Gregory, P. D., Naldini, L., & Bonini, C.
Affiliations
  • Vita-Salute San Raffaele University (IT)
  • The San Raffaele Telethon Institute for Gene Therapy (IT)
  • IRCCS Ospedale San Raffaele (IT)
  • Sangamo BioSciences (United States) (US)
  • University of Washington (US)
Corresponding author
  • Naldini, Luigi
  • Bonini, Chiara
ORCID
Venue
Nature Medicine
Publisher
Springer Science and Business Media LLC
Published
2012-04-01
Volume
18
Issue
5
Pages
807-815
ISSN
  • 1078-8956 (ISSN-L)
  • 1546-170X
DOI
10.1038/nm.2700
PMC
PMC5019824
OpenAlex
W2121950548
Cited by
448
References
0
Citations by year
2026: 17 · 2025: 18 · 2024: 14 · 2023: 22 · 2022: 34 · 2021: 27 · 2020: 40 · 2019: 40 · 2018: 39 · 2017: 44 · 2016: 30 · 2015: 45 · 2014: 27 · 2013: 37 · 2012: 13
FWCI
19.31
Type
journal article
Language
en
Open access
Licence
www.springer.com/tdm (tdm)
Locations
  • Nature Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Archivio Istituzionale della Ricerca - Università Vita-Salute San Raffaele (submittedVersion)
Funders
National Cancer Institute (P01CA018029)
Topics
  • Health Sciences › Medicine › Oncology › CAR-T cell therapy research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
Keywords
zinc finger nucleases, lentiviral transduction, leukemia immunotherapy, TCR gene transfer, WT1 antigen, T cell specificity, off-target toxicity, biosafety
MeSH terms
Base Sequence, Humans, Leukemia, Molecular Sequence Data, Receptors, Antigen, T-Cell, T-Lymphocytes, Deoxyribonucleases, Type II Site-Specific, Lentivirus, Zinc Fingers, Gene Transfer Techniques, Jurkat Cells, WT1 Proteins, T-Cell Antigen Receptor Specificity

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review