Abstract
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 AHNAK S2580F or ERBB2 H473Y or the HLA-DQB∗0601-restricted neoantigen ERBB2IP E805G 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.
| Original language | English |
|---|---|
| Pages (from-to) | 1078-1089 |
| Number of pages | 12 |
| Journal | Molecular Therapy |
| Volume | 24 |
| Issue number | 6 |
| DOIs | |
| State | Published - 1 Jun 2016 |
Bibliographical note
Funding Information:We would like to thank Perry Hackett (University of Minnesota) and Simon Olivares (M.D. Anderson Cancer Center) for their assistance with the Sleeping Beauty system. This research was supported by the Intramural Research Program of the NIH at the National Cancer Institute. On 7 May 2015, L.J.N.C. was appointed as the Chief Executive Officer at ZIOPHARM Oncology and remains at MD Anderson Cancer Center as a Visiting Scientist. The authors declare no other competing financial interests.
