Abstract 4095: Preclinical characterization of switchable allogeneic chimeric antigen receptor T cells to support first in human clinical study in CD123-positive hematologic and lymphatic malignancies
Authors: Ehninger Armin , Spehr Johannes , Loff Simon , Langer Anika , Riewaldt Julia , Reinhardt Julia , Meyer Jan-Erik , Dietrich Josephine , Jurado Gabriel , Lescarbeau Reynald , Cartellieri Marc 
Published: April 2023

Abstract
Approximately 80% of AML patients express CD123 on their leukemic blasts and CD123 is frequently expressed on other hematologic and lymphatic malignancies. Chimeric antigen receptor (CAR) T-cell therapy has demonstrated significant efficacy in B cell malignancies. Breakthrough of conventional CAR-T technology in AML has been hampered by expression of suitable target antigens on normal hematopoietic progenitor cells posing a risk for continued aplasia. Thus, innovative approaches putting the power of CAR-T technology under the control of reliable and fast-acting on/off switches to avoid and/or abrogate acute and long-term side effects are required. We have recently reported clinical proof-of-concept for autologous switchable CAR-T cell therapy in AML (Wermke et al. Blood 2021). However, manufacturing of autologous products is very costly and rrAML patients are often in urgent need for a product and multiple prior lines of treatment may be detrimental for product quality. While current clinical allogeneic approaches successfully prevent graft-versus-host disease, observed persistence has been limited due to rejection of the grafted T cells by host immune cells. Here, we report preclinical development of a donor-derived allogeneic switchable CAR-T therapy for CD123-positive hematologic and lymphatic malignancies (AVC-201). The reverse universal chimeric antigen receptor platform (RevCAR) is a 2-component CAR-T platform. The first component is a universal CAR-T cell. Its binding domain is a short, non-immunogenic peptide that by itself does not recognize any human cell surface antigen but is specifically bound by an scFv included in the second component, a soluble adaptor called targeting module (TM). To target CD123, a TM with a short half-life was selected (R-TM123), enabling a rapid switch-off of the RevCAR system by TM withdrawal to avoid acute and long-term toxicity usually associated with continuous activation of CAR-T cells. The allogeneic cells are generated by three edits with CRISPR-Cas9, which is meant to fully overcome graft-versus-host disease as well as graft rejection by host T and NK cells. Therefore, we expect a persistent T cell product that can be re-expanded with additional TM cycles. A summary of preclinical characterization of AVC-201 will be presented at the meeting including in vitro and in vivo pharmacology and toxicology experiments. Activation of Allo-RevCAR-T cells is strictly dependent on the presence of CD123-positive target cells and R-TM123. Allo-RevCAR-T cells redirected by R-TM123 efficiently lyse CD123-positive AML cells in vitro and in vivo, and in vitro EC50 values are in the low picomolar range of R-TM123 concentrations. In conclusion, preclinical data support the clinical exploration of AVC-201 in a first in human study.
Citation Format: Armin Ehninger, Johannes Spehr, Simon Loff, Anika Langer, Julia Riewaldt, Julia Reinhardt, Jan-Erik Meyer, Josephine Dietrich, Gabriel Jurado, Reynald Lescarbeau, Marc Cartellieri. Preclinical characterization of switchable allogeneic chimeric antigen receptor T cells to support first in human clinical study in CD123-positive hematologic and lymphatic malignancies. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4095.