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FAP is critical for ovarian cancer cell survival by sustaining NF-κB activation through recruitment of PRKDC in lipid rafts

Cancer Gene Therapy volume 30, pages 608–621 (2023)Cite this article

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Abstract

Fibroblast activation protein (FAP) is tumor-specific and plays an important role in tumorigenecity. However, agents against its enzymatic activity or extracellular presence were unsuccessful in the clinic for undefined reasons. Here we show that FAP expression is higher in advanced ovarian cancer and is only detected in invasive ovarian cancer cells. Silencing FAP induces apoptosis and FAP’s enzymatic activity is dispensable for cell survival. To elucidate the cause of apoptosis, we find that NF-κB activity is diminished when FAP is depleted and BIRC5 (survivin) acts downstream of FAP-NF-κB axis to promote cell survival. To uncover the link between FAP and NF-κB activation, we reveal that PRKDC (DNA-PK, DNA-dependent protein kinase) forms complex with FAP and is required for NF-κB activation and cell survival. Remarkably, FAP-PRKDC interaction occurs only in lipid rafts, and depleting FAP prevents lipid raft localization of PRKDC. Given the known ability of PRKDC to direct NF-κB activation, these results suggest that FAP recruits PRKDC in lipid rafts for NF-κB activation. FAP’s non-enzymatic role and functioning from lipid rafts for cell survival also offer an explanation on the failure of past FAP-targeted therapies. Finally, we demonstrate that EpCAM aptamer-delivered FAP siRNA impeded intraperitoneal xenograft development of ovary tumors.

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Fig. 1: Higher FAP expression is associated with advanced ovarian cancer.
Fig. 2: Knockdown of FAP leads to apoptosis of invasive ovarian cancer cells.
Fig. 3: NF-κB signaling is impaired in ovarian cancer cells with FAP-knockdown.
Fig. 4: BIRC5 is downstream of FAP-NF-κB axis regulating ovarian cancer cell survival.
Fig. 5: PRKDC is critically involved in NF-κB activation in ovarian cancer cells.
Fig. 6: FAP-PRKDC interaction occurs in lipid rafts of ovarian cancer cells.
Fig. 7: EpCAM aptamer-FAP siRNA chimera suppresses intraperitoneal xenograft development.

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All data generated or analysed during this study are included in this published article [and its supplementary information files].

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Acknowledgements

This is part of doctoral work of Bin Li at University of Florida College of Medicine. This work was supported by funding from NIH CA222467 (SH) and CA256482 (SH/LJ).

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  1. These authors contributed equally: Bin Li, Zuo Ding.

Authors and Affiliations

  1. Department of Anatomy and Cell Biology, University of Florida College of Medicine, Gainesville, FL, 32611, USA

    Bin Li, Zuo Ding, Ozlem Calbay, Yue Li, Tao Li & Shuang Huang

  2. Deparment of Molecular Medicine, The University of Texas Health Science Center at San Antonio, San Antonio, TX, 78245, USA

    Lingtao Jin

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BL, ZD, OC, and TL performed research and analyzed results; BL, ZD, and YL performed bioinformatics analysis; LJ provided experimental assistance and discussed results; SH designed research, supervised this study, and wrote the manuscript.

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Correspondence to Shuang Huang.

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Li, B., Ding, Z., Calbay, O. et al. FAP is critical for ovarian cancer cell survival by sustaining NF-κB activation through recruitment of PRKDC in lipid rafts. Cancer Gene Ther 30, 608–621 (2023). https://doi.org/10.1038/s41417-022-00575-x

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