Abstract B19: FAP CAR T cell therapy for solid tumors with PET imaging
Authors: Noguera-Ortega Estela , Lee Iris K , Xiao Zebin , Todd Leslie , Scholler John , Song Decheng , Liousia Maria , Lohith Katheryn , Xu Kexiang , Eduards Kimberly J , Farwell Michael D , June Carl H , Albelda Steven M , Puré Ellen , Sellmyer Mark A 
Published: December 2022

Two of the major obstacles that adoptive cell transfer immunotherapy needs to overcome to be successful in solid tumors are 1) the immunosuppressive tumor microenvironment (TME) and 2) the lack of robust biomarkers that allow for the identification of patients that would benefit from the therapy and for monitoring of the treatment response. Here we paired a CAR T cell therapy with a companion Positron Emission Tomography (PET) imaging approach that allows for serial, non-invasive, whole-body visualization of a biological target of interest -in this specific case, Fibroblast Activating Protein (FAP). FAP is a cell surface serine protease that is highly expressed by cancer associated fibroblasts in the TME that participate in the generation of the immunosuppressive stromagenic response in solid tumors. We designed a novel FAP CAR construct based on the scFv of the 4G5 antibody developed, in house, against canine FAP that cross-reacts against mouse and human FAP. In this study, we utilized the 18F-radiolabeled FAP inhibitor (FAPI), [18F]AlF-FAPI-74, to image FAP in two different mouse tumor models. First, the probe specificity was evaluated in the I45 human mesothelioma tumor model (a line which does not induce FAP+ fibroblasts). I45 WT and I45 cells transduced with human FAP were injected s.c. into the opposite flanks of a mouse and imaged following 2 weeks of tumor growth. We observed a 7.5-fold higher uptake of [18F]AlF-FAPI-74 in the I45 huFAP tumor compared to the WT, demonstrating the high specificity of the probe for FAP. Next, we used a more clinically-relevant A549 model where the tumor cells do not express FAP but induce a stromagenic response and drive the recruitment of FAP+ stromal cells in the TME. [18F]AlF-FAPI-74 PET/CT following 3 weeks of tumor growth showed a 6.5-fold increased radiotracer uptake in the tumor relative to the muscle at the baseline scan. Using this model, we evaluated the potential of our new CAR T cells to reduce tumor burden, as well as evaluating the potential of the [18F]AlF-FAPI-74 tracer as a tool to monitor the clearance of FAP-expressing cells in response to FAP CAR T cell therapy. Immediately after the baseline scan, we injected 5x106 FAP CAR T cells iv. At day 14 post-T cell injection, mice treated with FAP CAR T cells had significantly smaller tumors relative to the control group, which were treated with T cells that do not express the CAR, highlighting the therapeutic efficacy of the FAP CAR T cell therapy. Moreover, [18F]AlF-FAPI-74 PET/CT imaging showed no detectable tracer uptake in the tumors treated with FAP targeted CAR T. These findings were confirmed by immunofluorescence, indicating successful clearance of the FAP+ stroma by the injected FAP CAR T cells. In conclusion, the new 4G5 FAP CAR shows specific targeting toward mouse stroma infiltrating lung adenocarcinoma xenografts. PET imaging of FAP could be a highly useful approach to stratify patients prior to FAP CAR T therapy, as well as to monitor the pharmacodynamic response for FAP-targeted therapies.

Citation Format: Estela Noguera-Ortega, Iris K Lee, Zebin Xiao, Leslie Todd, John Scholler, Decheng Song, Maria Liousia, Katheryn Lohith, Kexiang Xu, Kimberly J Eduards, Michael D Farwell, Carl H June, Steven M Albelda, Ellen Puré, Mark A Sellmyer. FAP CAR T cell therapy for solid tumors with PET imaging [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy; 2022 Oct 21-24; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2022;10(12 Suppl):Abstract nr B19.