A novel TCR-like CAR with specificity for PR1/HLA-A2 effectively targets myeloid leukemia in vitro when expressed in human adult peripheral blood and cord blood T cells

作者：Ma Qing; Garber Haven R; Lu Sijie; He Hong; Tallis Eran; Ding Xiaoling; Sergeeva Anna; Wood Michael S; Dotti Gianpietro; Salvado Barbara; Ruisaard Kathryn; Clise Dwyer Karen; St John Lisa; Rezvani Katayoun; Alatrash Gheath; Shpall Elizabeth J; Molldrem Jeffrey J

来源：Cytotherapy, 2016, 18(8): 985-994.

DOI：10.1016/j.jcyt.2016.05.001

• acute myeloid leukemia (AML)
• CAR-T cell
• h8F4
• PR1
• UCB

Background aims. The PR1 peptide, derived from the leukemia-associated antigens proteinase 3 and neutrophil elastase, is overexpressed on HLA-A2 in acute myeloid leukemia (AML). We developed a T-cell receptor (TCR)-like monoclonal antibody (8F4) that binds the PR1/HLA-A2 complex on the surface of AML cells, efficiently killing them in vitro and eliminating them in preclinical models. Humanized 8F4 (h8F4) with high affinity for the PR1/HLA-A2 epitope was used to construct an h8F4-chimeric antigen receptor (CAR) that was transduced intoT cells to mediate anti-leukemia activity. Methods. Human T cells were transduced to express the PR1/HLA-A2-specific CAR (h8F4-CAR-T cells) containing the scFv of h8F4 fused to the intracellular signaling endo-domain of CD3 zeta chain through the transmembrane and intracellular costimulatory domain of CD28. Results. Adult human normal peripheral blood (PB) T cells were efficiently transduced with the h8F4CAR construct and predominantly displayed an effector memory phenotype with a minor population (12%) of central memory cells in vitro. Umbilical cord blood (UCB) T cells could also be efficiently transduced with the h8F4-CAR. The PB and UCB-derived h8F4-CAR-T cells specifically recognized the PR1/HLA-A2 complex and were capable of killing leukemia cell lines and primary AML blasts in an HLA-A2-dependent manner. Conclusions. Human adult PB and UCB-derived T cells expressing a CAR derived from the TCR-like 8F4 antibody rapidly and efficiently kill AML in vitro. Our data could lead to a new treatment paradigm for AML in which targeting leukemia stem cells could transfer long-term immunity to protect against relapse.

• 出版日期2016-8