Abstract 2321: Dual-function of CD27-CD70 costimulatory signal in CAR T cell therapy

Abstract Purpose: To investigate the function of co-stimulatory molecule CD27 on the antitumor efficacy of chimeric antigen receptor (CAR) T cells. Experimental Design: IL13 receptor α2 (IL13Rα2)-targeted CAR T cell products from glioblastoma (GBM) patients were characterized for CD27 surface expression and proliferation potential against recursive tumor challenge. Isolated CD27+ and CD27- fractions of CAR T cells were then evaluated for their antitumor potency against orthotopic GBM xenografts, as well as their molecular characteristics before and after tumor stimulation. Further, T cells were engineered to express a CAR construct together with a constitutively-expressed CD27 to investigate the role of continuous CD27 costimulation on CAR T cell maintenance and effector potency. Results: CD27 expression on CAR T cell products was correlated with their proliferation capacity against recursive tumor stimulation. Isolated CD27+ CAR T cells outperformed CD27- and unsorted cells in mediating long-term tumor eradication. CD27+ CAR T cells exhibited memory-associated genetic signatures, and were less exhausted following in vitro and in vivo tumor stimulation, both phenotypically and functionally. Moreover, CD27 also sensitized CAR T cells to target low antigen-expressing tumors. When targeting tumor cells expressing CD70 (CD27 ligand), CD27 on CAR T cells became rapidly down-regulated, suggesting a transient CD27-CD70 interaction. The advantageous effector function of CD27+ CAR T cells was diminished by blocking the CD27-CD70 interaction with CD70-targeting antibodies. CD70 is also expressed on CAR T cells, displaying a mutually exclusive expression pattern with CD27. Extended in vitro culture resulted in a phenotypic switch of CAR T cells from CD27+CD70- to CD27-CD70+. Surprisingly, constitutive expression of CD27 (CD27-cons) greatly impaired the effector activity of CAR T cells. These cells were able to eliminate tumor cells in vitro, but failed to mediate potent antitumor effect in vivo. Further characterization showed an early exhaustion phenotype for CD27-cons CAR T cells during in vitro expansion. Additionally, CD27-cons CAR T cells were also prone to activation-induced T cell apoptosis especially against high antigen-expressing targets. Conclusion: These studies demonstrate a dual-function of CD27 costimulation for CAR T cell antitumor activity. In the enriched CD27+ CAR T cell subset, the transient CD27-CD70 interaction with tumor cells resulted in enhanced antitumor potency. By contrast, constitutively expressed CD27 drove CAR T cell exhaustion and induced apoptosis. These results illustrate the critical role of CD27 in mediating potent CAR T cell therapeutic efficacy, and suggest the potential of CD27 as a marker for the functional quality of CAR T cell products. Citation Format: Dongrui Wang, Alfonso Brito, Darya Alizadeh, Renate Starr, Brenda Aguilar, Behnam Badie, Stephen J. Forman, Christine E. Brown. Dual-function of CD27-CD70 costimulatory signal in CAR T cell therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2321.