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

• Published in: Haematologica (Roma) 2010-01, Vol.95 (1), p.126-134
• Main Authors: XUE, Shao-An, LIQUAN GAO, THOMAS, Sharyn, HART, Daniel P, ZHAO XUE, John, GILLMORE, Roopinder, VOSS, Ralf-Holger, MORRIS, Emma, STAUSS, Hans J
• Format: Article
• Language: English
• Subjects: Adult; Animals; Antigens, Neoplasm - genetics; Antigens, Neoplasm - immunology; Antigens, Neoplasm - metabolism; Biological and medical sciences; Blast Crisis - genetics; Blast Crisis - immunology; Blast Crisis - therapy; Genetic Engineering - methods; Genetic Therapy - methods; Genetic Vectors - biosynthesis; Genetic Vectors - chemistry; Hematologic and hematopoietic diseases; Hepatitis B Virus, Woodchuck - genetics; Humans; Jurkat Cells; Kidneys; Leukemia - genetics; Leukemia - immunology; Leukemia - therapy; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Medical sciences; Mice; Mice, Inbred NOD; Mice, SCID; Nephrology. Urinary tract diseases; Original; Receptors, Antigen, T-Cell - genetics; Receptors, Antigen, T-Cell - physiology; Receptors, Antigen, T-Cell - therapeutic use; T-Lymphocytes - immunology; T-Lymphocytes - metabolism; T-Lymphocytes - pathology; Transplantation, Autologous - immunology; Tumors of the urinary system; Wilms Tumor - immunology; Wilms Tumor - pathology; Wilms Tumor - therapy
• ISSN: 0390-6078; 1592-8721
• DOI: 10.3324/haematol.2009.006486

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. 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. 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. 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.