RT Journal Article
SR Electronic
T1 Targeting the extra domain A of fibronectin for cancer therapy with CAR-T cells
JF Journal for ImmunoTherapy of Cancer
JO J Immunother Cancer
FD BMJ Publishing Group Ltd
SP e004479
DO 10.1136/jitc-2021-004479
VO 10
IS 8
A1 Celia Martín-Otal
A1 Aritz Lasarte-Cia
A1 Diego Serrano
A1 Noelia Casares
A1 Enrique Conde
A1 Flor Navarro
A1 Inés Sánchez-Moreno
A1 Marta Gorraiz
A1 Patricia Sarrión
A1 Alfonso Calvo
A1 Carlos E De Andrea
A1 José Echeveste
A1 Amaia Vilas
A1 Juan Roberto Rodriguez-Madoz
A1 Jesús San Miguel
A1 Felipe Prosper
A1 Sandra Hervas-Stubbs
A1 Juan Jose Lasarte
A1 Teresa Lozano
YR 2022
UL http://jitc.bmj.com/content/10/8/e004479.abstract
AB Background One of the main difficulties of adoptive cell therapies with chimeric antigen receptor (CAR)-T cells in solid tumors is the identification of specific target antigens. The tumor microenvironment can present suitable antigens for CAR design, even though they are not expressed by the tumor cells. We have generated a CAR specific for the splice variant extra domain A (EDA) of fibronectin, which is highly expressed in the tumor stroma of many types of tumors but not in healthy tissues.Methods EDA expression was explored in RNA-seq data from different human tumor types and by immunohistochemistry in paraffin-embedded tumor biopsies. Murine and human anti-EDA CAR-T cells were prepared using recombinant retro/lentiviruses, respectively. The functionality of EDA CAR-T cells was measured in vitro in response to antigen stimulation. The antitumor activity of EDA CAR-T cells was measured in vivo in C57BL/6 mice challenged with PM299L-EDA hepatocarcinoma cell line, in 129Sv mice-bearing F9 teratocarcinoma and in NSG mice injected with the human hepatocarcinoma cell line PLC.Results EDA CAR-T cells recognized and killed EDA-expressing tumor cell lines in vitro and rejected EDA-expressing tumors in immunocompetent mice. Notably, EDA CAR-T cells showed an antitumor effect in mice injected with EDA-negative tumor cells lines when the tumor stroma or the basement membrane of tumor endothelial cells express EDA. Thus, EDA CAR-T administration delayed tumor growth in immunocompetent 129Sv mice challenged with teratocarcinoma cell line F9. EDA CAR-T treatment exerted an antiangiogenic effect and significantly reduced gene signatures associated with epithelial-mesenchymal transition, collagen synthesis, extracellular matrix organization as well as IL-6-STAT5 and KRAS pathways. Importantly, the human version of EDA CAR, that includes the human 41BB and CD3ζ endodomains, exerted strong antitumor activity in NSG mice challenged with the human hepatocarcinoma cell line PLC, which expresses EDA in the tumor stroma and the endothelial vasculature. EDA CAR-T cells exhibited a tropism for EDA-expressing tumor tissue and no toxicity was observed in tumor bearing or in healthy mice.Conclusions These results suggest that targeting the tumor-specific fibronectin splice variant EDA with CAR-T cells is feasible and offers a therapeutic option that is applicable to different types of cancer.Data are available in a public, open access repository. Data are available on reasonable request. The expression profile data analyzed in this study are available from Gene Expression Omnibus (GSE204887).