Abstract 4053: A mesothelin targeting chimeric antigen receptor macrophage (CAR-M) for solid tumor immunotherapy: pre-clinical development of CT-1119
Authors: Anderson Nicholas R. , Shah Brinda , Worth Alison , Gabbasov Rashid , Menchel Brett , Ciccaglione Kerri , Blumenthal Daniel , Pierini Stefano , DeLong Sabrina Ceeraz , Abramson Sascha , Condamine Thomas , Klichinsky Michael 
Published: April 2023

Abstract
While adoptive cell therapies have seen significant success in the treatment of hematological malignancies, solid tumors remain challenging for the field. A significant obstacle is the exclusion of T cells from the tumor microenvironment (TME). In contrast, monocytes/macrophages are naturally recruited to the TME. These cells then have the potential to phagocytose tumor cells, activate the TME, and prime a broad anti-tumor adaptive immune response via T cell recruitment and activation. We have previously developed CT-0508, a chimeric antigen receptor macrophage (CAR-M) targeting HER2 which showed efficacy in a variety of pre-clinical models and is currently in a Phase I clinical trial for patients with HER2+ solid tumors. Mesothelin is overexpressed in a variety of solid tumors, including mesothelioma, lung, pancreatic, and ovarian cancers. To leverage tumor biology with myeloid cells, we engineered primary human macrophages using the chimeric adenoviral vector Ad5f35 to express a CAR containing a human scFv against human mesothelin. We used both in vitro cell based assays and in vivo xenograft models to assess the activity of CT-1119. CAR-M engineered with an Ad5f35 vector demonstrated high CAR expression, high viability, upregulated M1 (anti-tumor) macrophage markers, and downregulated M2 (pro-tumor) macrophage markers. CT-1119 specifically phagocytosed multiple mesothelin expressing tumor cell lines in a CAR-dependent and antigen-dependent manner. CT-1119 demonstrated robust in vitro killing of the relevant tumor cell lines A549 and MES-OV expressing mesothelin. CAR engagement also induced the release of pro-inflammatory cytokines such as TNFα following stimulation with mesothelin in both cell-free and cell-based contexts in a dose-dependent manner. In vivo, CT-1119 significantly reduced tumor burden in a murine xenograft model of lung cancer. Similarly, human monocytes targeting mesothelin were successfully generated using the same Ad5f35 vector and demonstrated specific activity against mesothelin positive tumor cells. The presented results demonstrate that CT-1119, an autologous human anti-mesothelin CAR-M, can cause phagocytosis, tumor cell killing, and pro-inflammatory cytokine release in response to stimulation with mesothelin. These results show that CAR-M is a feasible approach for the treatment of mesothelin expressing sold tumors via the potential for induction of a systemic anti-tumor response.
Citation Format: Nicholas R. Anderson, Brinda Shah, Alison Worth, Rashid Gabbasov, Brett Menchel, Kerri Ciccaglione, Daniel Blumenthal, Stefano Pierini, Sabrina Ceeraz DeLong, Sascha Abramson, Thomas Condamine, Michael Klichinsky. A mesothelin targeting chimeric antigen receptor macrophage (CAR-M) for solid tumor immunotherapy: pre-clinical development of CT-1119. [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 4053.