NY-ESO-1-specific redirected T cells with endogenous TCR knockdown mediate tumor response and cytokine release syndrome

Abstract

BackgroundBecause of the shortage of ideal cell surface antigens, the development of T-cell receptor (TCR)-engineered T cells (TCR-T) that target intracellular antigens such as NY-ESO-1 is a promising approach for treating patients with solid tumors. However, endogenous TCRs in vector-transduced T cells have been suggested to impair cell-surface expression of transduced TCR while generating mispaired TCRs that can become self-reactive.MethodsWe conducted a first-in-human phase I clinical trial with the TCR-transduced T-cell product (TBI-1301) in patients with NY-ESO-1-expressing solid tumors. In manufacturing TCR-T cells, we used a novel affinity-enhanced NY-ESO-1-specific TCR that was transduced by a retroviral vector that enables siRNA (small interfering RNA)-mediated silencing of endogenous TCR. The patients were divided into two cohorts. Cohort 1 was given a dose of 5×108 cells (whole cells including TCR-T cells) preconditioned with 1500 mg/m2 cyclophosphamide. Cohort 2 was given 5× 109 cells preconditioned with 1500 mg/m2 cyclophosphamide.ResultsIn vitro study showed that both the CD8+ and CD4+ T fractions of TCR-T cells exhibited cytotoxic effects against NY-ESO-1-expressing tumor cells. Three patients and six patients were allocated to cohort 1 and cohort 2, respectively. Three of the six patients who received 5×109 cells showed tumor response, while three patients developed early-onset cytokine release syndrome (CRS). One of the patients developed a grade 3 lung injury associated with the infiltration of the TCR-T cells. No siRNA-related adverse events other than CRS were observed. Cytokines including interleukin 6 I and monocyte chemotactic protein-1/chemokine (C-C motif) ligand (CCL2)increased in the sera of patients with CRS. In vitro analysis showed these cytokines were not secreted from the T cells infused. A significant fraction of the manufactured T cells in patients with CRS was found to express either CD244, CD39, or both at high levels.ConclusionsThe trial showed that endogenous TCR-silenced and affinity-enhanced NY-ESO-1 TCR-T cells were safely administered except for grade 3 lung injury. The TCR-T cell infusion exhibited significant tumor response and early-onset CRS in patients with tumors that express NY-ESO-1 at high levels. The differentiation properties of the manufactured T cells may be prognostic for TCR-T-related CRS.Trial registration numberNCT02366546.

Coengineering specificity, safety, and function into T cells for cancer immunotherapy

“…Notably, one of our dual β-chain amino acid replacement TCR designs (DMβ; along with endogenous TCR knockdown) has been translated to the clinic by others and mediated tumor response. 175 Taken together, it is evident that binding-enhanced TCRs can augment T-cell function but beyond an affinity-threshold activity levels are abrogated and specificity is lost (Figure 4A). Presumably, the optimal affinity range will vary from TCR to TCR.…”

“…NY‐ESO‐1 is expressed by a broad range of cancers including synovial sarcoma (as mentioned above), melanoma, and epithelial ovarian cancer 169,170 but in healthy adult tissues it is restricted to male germ cells 171 . A2/NY TCRs have demonstrated safety, persistence, and anti‐tumor activity in the clinic (e.g., against melanoma and sarcoma), including TCRs that have been affinity‐enhanced 140,172–175 . The BC1 TCR is near identical in sequence to the well‐studied 1G4 TCR, and crystal structures of 1G4 and 1G4 variants, in complex with A2/NY have been solved 176,177 .…”

“…171 A2/ NY TCRs have demonstrated safety, persistence, and anti-tumor activity in the clinic (e.g., against melanoma and sarcoma), including TCRs that have been affinity-enhanced. 140,[172][173][174][175] The BC1 TCR is near identical in sequence to the well-studied 1G4 TCR, and crystal structures of 1G4 and 1G4 variants, in complex with A2/NY have been solved. 176,177 Previously, by structure-based computational design (using the crystal coordinates for 1G4:A2/NY, 2BNR in the Protein Databank) and free-energy calculations 178 we (with Profs.…”

“…We have since demonstrated significantly improved tumor control (against A2 + /NY + melanoma Me275 and A375) by T cells expressing A2/NY TCRs in the upper limit of natural affinity (e.g., TCRs DMβ and A97L) 181 as compared to wild‐type, and no benefit upon ACT of T cells bearing the supraphysiologic affinity TCR wtc51m (Stefanidis et al, in revision, Semilietof et al, in preparation). Notably, one of our dual β‐chain amino acid replacement TCR designs (DMβ; along with endogenous TCR knockdown) has been translated to the clinic by others and mediated tumor response 175 …”

T cell receptor gene‐modified allogeneic T cells with siRNA for endogenous T cell receptor induce efficient tumor regression without graft‐versus‐host disease

“…18 Human lymphocytes transduced with MS3-NY-ESO-1-siTCR showed NY-ESO-1-specific cytotoxicity in human tumor cells. 22 To target β-2 microglobulin, which is required for the expression of HLA class I, we created a guide RNA (GAGTA GCG CGA GCA CAG CTAAGG) that was ligated into the pLVSIN-EF1α (Takara Bio) lentiviral vector.…”