Stable, Nonviral Expression of Mutated Tumor Neoantigen-specific T-cell Receptors Using the Sleeping Beauty Transposon/Transposase System

• Published in: Molecular therapy 2016-06, Vol.24 (6), p.1078-1089
• Main Authors: Deniger, Drew C, Pasetto, Anna, Tran, Eric, Parkhurst, Maria R, Cohen, Cyrille J, Robbins, Paul F, Cooper, Laurence JN, Rosenberg, Steven A
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antigens; Cancer therapies; Cholangiocarcinoma; Clinical trials; Cloning; Cytokines; DNA Transposable Elements; Gene therapy; Genetic Engineering; HLA-A2 Antigen - metabolism; HLA-DQ beta-Chains - metabolism; Humans; Lymphocytes; Medicin och hälsovetenskap; Melanoma; Membrane Proteins - immunology; Metastasis; Mice; Mutation; Neoplasms - immunology; Original; Patients; Peptides; Plasmids; Pneumoviridae; Receptor, ErbB-2 - immunology; Receptors, Antigen, T-Cell - metabolism; T-Lymphocytes - immunology; T-Lymphocytes - pathology; Transposases - metabolism; Tumors
• ISSN: 1525-0016; 1525-0024; 1525-0024
• DOI: 10.1038/mt.2016.51

Neoantigens unique to each patient's tumor can be recognized by autologous T cells through their T-cell receptor (TCR) but the low frequency and/or terminal differentiation of mutation-specific T cells in tumors can limit their utility as adoptive T-cell therapies. Transfer of TCR genes into younger T cells from peripheral blood with a high proliferative potential could obviate this problem. We generated a rapid, cost-effective strategy to genetically engineer cancer patient T cells with TCRs using the clinical Sleeping Beauty transposon/transposase system. Patient-specific TCRs reactive against HLA-A*0201-restriced neoantigens AHNAKS2580F or ERBB2H473Y or the HLA-DQB*0601-restricted neoantigen ERBB2IPE805G were assembled with murine constant chains and cloned into Sleeping Beauty transposons. Patient peripheral blood lymphocytes were coelectroporated with SB11 transposase and Sleeping Beauty transposon, and transposed T cells were enriched by sorting on murine TCRβ (mTCRβ) expression. Rapid expansion of mTCRβ+ T cells with irradiated allogeneic peripheral blood lymphocytes feeders, OKT3, interleukin-2 (IL-2), IL-15, and IL-21 resulted in a preponderance of effector (CD27−CD45RA−) and less-differentiated (CD27+CD45RA+) T cells. Transposed T cells specifically mounted a polyfunctional response against cognate mutated neoantigens and tumor cell lines. Thus, Sleeping Beauty transposition of mutation-specific TCRs can facilitate the use of personalized T-cell therapy targeting unique neoantigens.