Development of a Wilms' tumor antigen-specific T-cell receptor for clinical trials: engineered patient's T cells can eliminate autologous leukemia blasts in NOD/SCID mice.
BACKGROUND
The Wilms' tumor antigen (WT1) is an attractive target for immunotherapy of leukemia. In the past, we isolated and characterized the specificity and function of a WT1-specific T-cell receptor. The goal of this translational study was to develop a safe and efficient WT1-T-cell receptor retroviral vector for an adoptive immunotherapy trial with engineered T cells.


METHODS
We generated a panel of retroviral constructs containing unmodified or codon-optimized WT1-T-cell receptor alpha and beta genes, linked via internal ribosome entry sites or 2A sequences, with or without an additional inter-chain disulfide bond in the T-cell receptor constant domains. These constructs were functionally analyzed in vitro, and the best one was tested in an autologous primary leukemia model in vivo.


RESULTS
We identified a WT1-T-cell receptor construct that showed optimal tetramer staining, antigen-specific cytokine production and killing activity when introduced into primary human T cells. Fresh CD34(+) cells purified from a patient with leukemia were engrafted into NOD/SCID mice, followed by adoptive immunotherapy with patient's autologous T cells transduced with the WT1-T-cell receptor. This therapeutic treatment evidently decreased leukemia engraftment in mice and resulted in a substantial improvement of leukemia-free survival.


CONCLUSIONS
This is the first report that patient's T cells, engineered to express the WT1-T-cell receptor, can eliminate autologous leukemia progenitor cells in an in vivo model. This study provides a firm basis for the planned WT1-T-cell receptor gene therapy trial in leukemia patients.
Xue
Shao-An
SA
1Department of Immunology, Division of Infection & Immunity, University College London, Royal Free Hospital, Rowland Hill Street,London, UK. s.xue@medsch.ucl.ac.uk
Gao
Liquan
L
Thomas
Sharyn
S
Hart
Daniel P
DP
Xue
John Zhao
JZ
Gillmore
Roopinder
R
Voss
Ralf-Holger
RH
Morris
Emma
E
Stauss
Hans J
HJ
D000328:Adult; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D001752:Blast Crisis / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D005818:Genetic Engineering / Q000379:methods*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000096:biosynthesis / Q000737:chemistry; D017919:Hepatitis B Virus, Woodchuck / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells; D007938:Leukemia / Q000235:genetics / Q000276:immunology* / Q000628:therapy; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D011948:Receptors, Antigen, T-Cell / Q000235:genetics* / Q000502:physiology / Q000627:therapeutic use*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D014182:Transplantation, Autologous / Q000276:immunology; D009396:Wilms Tumor / Q000276:immunology / Q000473:pathology* / Q000628:therapy
D000951:Antigens, Neoplasm; D011948:Receptors, Antigen, T-Cell
Nat Biotechnol. 2004 May;22(5):589-94
15064769
Eur J Haematol. 2009 Jan;82(1):61-8
18801058
Nat Rev Immunol. 2008 Apr;8(4):247-58
18323851
Leukemia. 1995 Jun;9(6):1060-7
7596170
Blood. 1996 Sep 15;88(6):2267-78
8822948
Blood. 2007 Mar 15;109(6):2331-8
17082316
Bone Marrow Transplant. 2004 Oct;34(7):653-4
15258561
Haematologica. 2005 Oct;90(10):1415-21
16219579
Blood. 1997 Feb 15;89(4):1405-12
9028964
Leukemia. 1999 Mar;13(3):393-9
10086730
Blood. 2008 Jan 1;111(1):236-42
17875804
Proc Natl Acad Sci U S A. 2004 Sep 21;101(38):13885-90
15365188
Eur J Haematol. 2008 Mar;80(3):201-7
18081724
Science. 2002 Oct 25;298(5594):850-4
12242449
Blood. 2000 Apr 1;95(7):2198-203
10733485
Blood. 2003 Mar 1;101(5):1698-704
12406915
Blood. 1994 Nov 1;84(9):3071-9
7949179
Blood. 1996 Dec 1;88(11):4396-8
8943879
Blood. 1997 Aug 1;90(3):1217-25
9242555
Blood. 2005 Nov 1;106(9):3062-7
16020516
Leukemia. 2004 Jan;18(1):165-6
14603333
Leukemia. 2008 Sep;22(9):1721-7
18548092
Clin Cancer Res. 2006 Apr 15;12(8):2434-41
16638849
Immunogenetics. 2000 Feb;51(2):99-107
10663572
Br J Haematol. 2004 Jun;125(5):590-600
15147374
Gene Ther. 2005 Jan;12(1):3-4
15510172
Clin Cancer Res. 2005 Dec 15;11(24 Pt 1):8799-807
16361568
Clin Cancer Res. 2006 Jan 1;12(1):34-42
16397021
Int J Cancer. 2006 Sep 15;119(6):1360-7
16596644
Acta Haematol. 2004;112(1-2):79-84
15179007
Clin Immunol. 2006 May;119(2):135-45
16458072
Proc Natl Acad Sci U S A. 2004 Oct 5;101 Suppl 2:14639-45
15381769
J Hepatol. 2001 Apr;34(4):562-9
11394656
ScientificWorldJournal. 2007 May 29;7:649-65
17619750
Science. 2006 Oct 6;314(5796):126-9
16946036
Blood. 2007 Sep 15;110(6):1924-32
17505014
Gene Ther. 2006 Apr;13(7):641-5
16355114
J Immunol. 2007 Nov 1;179(9):5803-10
17947653
Gene Ther. 2008 Apr;15(8):625-31
18305579