Abstract 3750: Micromolar affinity CAR T cells to ICAM-1 achieves rapid tumor elimination while avoiding systemic toxicity
Authors: Enda Shevlin, Spencer Park, Yogindra Vedvyas, Marjan Zaman, Susan Park, Irene M. Min, Moonsoo M. Jin
Journal: Cancer Research (2017)

Abstract Introduction: Adoptive immune therapy has achieved great success in eradicating blood-borne cancers, prominently, the CD19 CAR T cells in B cell leukemia and lymphomas. However, CAR T cell therapy in solid tumors has been limited due to the scarcity of tumor antigens that are deemed safe for targeting. One strategy to overcome scarcity of tumor antigen is by tuning the affinity of CAR to limit T cell reaction with cells overexpressing target antigen while sparing cells with basal level expression. To rigorously test the idea of “affinity tuning”, we built variants of CARs possessing one million-fold difference in affinity spanning low nanomolar to high micromolar affinity to a target antigen, and examined the influence of CAR affinity on the rate of tumor killing and systemic toxicity. Methods: Antigen-binding domain of CAR was built from the inserted or I domain belonging to integrin LFA-1. Affinity of CAR expressed in T cells was confirmed by ICAM-1 binding by flow cytometry. For in vivo study, mice with systemic growth of ICAM-1 positive thyroid tumor were used, where tumors grew in lungs, liver, and bones. Tumor growth and killing were monitored by whole body luminescence imaging. Sera were collected for cytokine analysis. Body weight, cytokine profile, and overall behavior were used to assess the severity of systemic toxicity. Results: CAR T cells with a step-wise, one million-fold variation in affinity to ICAM-1 resulted in a rate of target killing in proportion to the increase in affinity and in ICAM-1 density. Owing to cross-reaction of human LFA-1 with murine ICAM-1, the influence of CAR affinity on efficacy and on-target, off-tumor toxicity was tested in mice bearing ICAM-1 positive human tumors. In vivo tumor elimination by CAR T cells was in contrast to in vitro affinity-dependent rate of target killing, demonstrating that micromolar affinity CAR T cells was superior to nanomolar affinity T cells in both tumor killing and safety aspects. Highest affinity CAR T cells led to uniform death of the host, caused by on-target, off-tumor toxicity, and high level cytokine release. Conclusion: Our study is the first comprehensive report examining the effect of CAR affinity on the rate of tumor killing, efficacy, and toxicity. In contrast to in vitro tumor killing effect, the increase in affinity of CAR beyond certain threshold was deleterious to T cell persistence and associated with more frequent tumor relapse. Our study highlights that CAR T cells approximating natural T cell receptor affinity is more efficacious in eliminating tumors with overexpressed antigens, and is safer by avoiding potential reaction with normal cells with basal expression of the same antigen. Citation Format: Enda Shevlin, Spencer Park, Yogindra Vedvyas, Marjan Zaman, Susan Park, Irene M. Min, Moonsoo M. Jin. Micromolar affinity CAR T cells to ICAM-1 achieves rapid tumor elimination while avoiding systemic toxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3750. doi:10.1158/1538-7445.AM2017-3750