Related Experiment Video
Updated: Jul 2, 2025
Generation and Expansion of Primary, Malignant Pleural Mesothelioma Tumor Lines
Published on: April 21, 2022
Mesothelin CAR T Cells Secreting Anti-FAP/Anti-CD3 Molecules Efficiently Target Pancreatic Adenocarcinoma and its
Marc Wehrli1,2, Samantha Guinn3,4, Filippo Birocchi1,2
1Cellular Immunotherapy Program, Cancer Center, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts.
Purpose:
Targeting solid tumors with chimeric antigen receptor (CAR) T cells remains challenging due to heterogenous target antigen expression, antigen escape, and the immunosuppressive tumor microenvironment (TME). Pancreatic cancer is characterized by a thick stroma generated by cancer-associated fibroblasts (CAF), which may contribute to the limited efficacy of mesothelin-directed CAR T cells in early-phase clinical trials. To provide a more favorable TME for CAR T cells to target pancreatic ductal adenocarcinoma (PDAC), we generated T cells with an antimesothelin CAR and a secreted T-cell-engaging molecule (TEAM) that targets CAF through fibroblast activation protein (FAP) and engages T cells through CD3 (termed mesoFAP CAR-TEAM cells).
Experimental Design:
Using a suite of in vitro, in vivo, and ex vivo patient-derived models containing cancer cells and CAF, we examined the ability of mesoFAP CAR-TEAM cells to target PDAC cells and CAF within the TME. We developed and used patient-derived ex vivo models, including patient-derived organoids with patient-matched CAF and patient-derived organotypic tumor spheroids.
Results:
We demonstrated specific and significant binding of the TEAM to its respective antigens (CD3 and FAP) when released from mesothelin-targeting CAR T cells, leading to T-cell activation and cytotoxicity of the target cell. MesoFAP CAR-TEAM cells were superior in eliminating PDAC and CAF compared with T cells engineered to target either antigen alone in our ex vivo patient-derived models and in mouse models of PDAC with primary or metastatic liver tumors.
Conclusions:
CAR-TEAM cells enable modification of tumor stroma, leading to increased elimination of PDAC tumors. This approach represents a promising treatment option for pancreatic cancer.
Insights
Engineered T cells targeting both pancreatic cancer cells and their stroma show improved tumor elimination. This novel CAR-TEAM cell therapy offers a promising new treatment strategy for pancreatic ductal adenocarcinoma.
Area of Science:
- Oncology
- Immunotherapy
- Cancer Biology
Background:
- Chimeric antigen receptor (CAR) T cell therapy faces challenges in solid tumors like pancreatic cancer due to tumor microenvironment (TME) and antigen heterogeneity.
- Pancreatic ductal adenocarcinoma (PDAC) features a dense stroma rich in cancer-associated fibroblasts (CAF), hindering CAR T cell efficacy.
- Existing mesothelin-directed CAR T cells show limited success in early clinical trials for pancreatic cancer.
Purpose of the Study:
- To develop a novel CAR T cell strategy to overcome the immunosuppressive TME in pancreatic cancer.
- To engineer T cells with a chimeric antigen receptor (CAR) targeting mesothelin and a secreted T-cell-engaging molecule (TEAM) targeting cancer-associated fibroblasts (CAF) via fibroblast activation protein (FAP).
- To evaluate the efficacy of these dual-targeting mesoFAP CAR-TEAM cells in preclinical models of pancreatic cancer.
Main Methods:
- Utilized in vitro, in vivo, and ex vivo patient-derived models, including organoids and organotypic tumor spheroids, co-cultured with cancer cells and CAF.
- Developed mesoFAP CAR-TEAM cells designed to target both PDAC cells (via mesothelin) and CAF (via FAP).
- Assessed T cell activation, binding affinity, and cytotoxicity against both cancer and stromal cells.
Main Results:
- Demonstrated specific binding and activation of mesoFAP CAR-TEAM cells upon engagement with CD3 and FAP antigens.
- MesoFAP CAR-TEAM cells exhibited superior elimination of both PDAC cells and CAF compared to single-targeting CAR T cells.
- Enhanced efficacy was observed in both ex vivo patient-derived models and in vivo mouse models of PDAC, including liver metastases.
Conclusions:
- Engineered CAR-TEAM cells effectively modify the tumor stroma, leading to improved pancreatic cancer elimination.
- This dual-targeting strategy represents a promising therapeutic approach for pancreatic cancer.
- The mesoFAP CAR-TEAM cell therapy holds potential for enhancing treatment outcomes in pancreatic cancer patients.
Related Concept Videos
02:30Metastasis
Epithelial-to-Mesenchymal Transition
The epithelial-to-mesenchymal transition or EMT is a developmental process commonly observed in wound healing, embryogenesis, and cancer metastasis. EMT is induced by transforming growth factor-beta (TGF-β) or receptor tyrosine kinase (RTK) ligands, which further...
02:17The Tumor Microenvironment