Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma

Abstract This study investigates the therapeutic potential of c-Met-targeted CAR T cells co-expressing a truncated TGFβ receptor II (TGFBR2-N) to overcome TGFβ1-mediated immunosuppression in lung adenocarcinoma. Bioinformatics analysis using the GEPIA2 database and single-cell RNA sequencing (scRNA-...

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Main Authors: Chao Su, Wenbin Hu, Huafang Mao
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-15086-6
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author Chao Su
Wenbin Hu
Huafang Mao
author_facet Chao Su
Wenbin Hu
Huafang Mao
author_sort Chao Su
collection DOAJ
description Abstract This study investigates the therapeutic potential of c-Met-targeted CAR T cells co-expressing a truncated TGFβ receptor II (TGFBR2-N) to overcome TGFβ1-mediated immunosuppression in lung adenocarcinoma. Bioinformatics analysis using the GEPIA2 database and single-cell RNA sequencing (scRNA-seq) revealed high c-Met expression in lung adenocarcinoma and highlighted the role of TGFβ signaling in modulating tumor-infiltrating T cells. A CAR construct targeting c-Met was developed to co-express TGFBR2-N via lentiviral transduction, and CAR T cell functionality was assessed through IL-2 ELISA, flow cytometry for pSMAD2/3 signaling, CD69 and PD-1 expression, as well as proliferation and cytotoxicity assays. Immunohistochemistry and multiplex cytokine analysis demonstrated that TGFBR2-N co-expression reduced pSMAD2/3 signaling, neutralized TGFβ1’s suppressive effects, and enhanced CAR T cell proliferation and cytotoxicity. In vivo, TGFBR2-N co-expression promoted tumor suppression, increased CD3 + T cell infiltration, and elevated levels of IFN-γ, IL-1β, IL-6, and TNF-α in the tumor microenvironment. These findings suggest that co-expressing TGFBR2-N in c-Met CAR T cells counteracts TGFβ1-mediated immunosuppression, enhancing their therapeutic efficacy in lung adenocarcinoma and offering a promising strategy for improving CAR T cell therapy in solid tumors.
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spelling doaj-art-5fdf4264bcd54054b120e09852e13d542025-08-20T03:46:01ZengNature PortfolioScientific Reports2045-23222025-08-0115111010.1038/s41598-025-15086-6Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinomaChao Su0Wenbin Hu1Huafang Mao2Department of Cardiothoracic Surgery, Affiliated Hospital of Shaoxing University (The Shaoxing Municipal Hospital)Department of Cardiothoracic Surgery, Affiliated Hospital of Shaoxing University (The Shaoxing Municipal Hospital)Department of Cardiothoracic Surgery, Affiliated Hospital of Shaoxing University (The Shaoxing Municipal Hospital)Abstract This study investigates the therapeutic potential of c-Met-targeted CAR T cells co-expressing a truncated TGFβ receptor II (TGFBR2-N) to overcome TGFβ1-mediated immunosuppression in lung adenocarcinoma. Bioinformatics analysis using the GEPIA2 database and single-cell RNA sequencing (scRNA-seq) revealed high c-Met expression in lung adenocarcinoma and highlighted the role of TGFβ signaling in modulating tumor-infiltrating T cells. A CAR construct targeting c-Met was developed to co-express TGFBR2-N via lentiviral transduction, and CAR T cell functionality was assessed through IL-2 ELISA, flow cytometry for pSMAD2/3 signaling, CD69 and PD-1 expression, as well as proliferation and cytotoxicity assays. Immunohistochemistry and multiplex cytokine analysis demonstrated that TGFBR2-N co-expression reduced pSMAD2/3 signaling, neutralized TGFβ1’s suppressive effects, and enhanced CAR T cell proliferation and cytotoxicity. In vivo, TGFBR2-N co-expression promoted tumor suppression, increased CD3 + T cell infiltration, and elevated levels of IFN-γ, IL-1β, IL-6, and TNF-α in the tumor microenvironment. These findings suggest that co-expressing TGFBR2-N in c-Met CAR T cells counteracts TGFβ1-mediated immunosuppression, enhancing their therapeutic efficacy in lung adenocarcinoma and offering a promising strategy for improving CAR T cell therapy in solid tumors.https://doi.org/10.1038/s41598-025-15086-6Lung cancerChimeric antigen receptorT cellTGF-β
spellingShingle Chao Su
Wenbin Hu
Huafang Mao
Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
Scientific Reports
Lung cancer
Chimeric antigen receptor
T cell
TGF-β
title Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
title_full Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
title_fullStr Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
title_full_unstemmed Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
title_short Co-expression of a truncated TGFβ receptor II in c-Met CAR T cells enhances antitumor activity against lung adenocarcinoma
title_sort co expression of a truncated tgfβ receptor ii in c met car t cells enhances antitumor activity against lung adenocarcinoma
topic Lung cancer
Chimeric antigen receptor
T cell
TGF-β
url https://doi.org/10.1038/s41598-025-15086-6
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AT wenbinhu coexpressionofatruncatedtgfbreceptoriiincmetcartcellsenhancesantitumoractivityagainstlungadenocarcinoma
AT huafangmao coexpressionofatruncatedtgfbreceptoriiincmetcartcellsenhancesantitumoractivityagainstlungadenocarcinoma