Abstract B088: Development of a silk-alginate-based 3-dimensional tumor model to screen anti-cancer drugs: Targeting S1P signaling for oral cancer therapy

Abstract Background: Despite advancements in the diagnostic and treatment modalities, the 5-year survival rate of oral squamous cell carcinoma (OSCC) patients has not demonstrated considerable improvement due to a high rate of recurrence and chemo-therapy resistance. Therefore, there is a compelling need to search for new molecular drug- targets for oral cancer. Towards this, sphingosine-1-phosphate (S1P) signaling plays a pivotal role in oral carcinogenesis and chemo-resistance. Further, owing to the disadvantages associated with conventional two-dimensional screening methods, there has been a shift towards the application of in vitro 3-dimensional (3-D) tumor models which can faithfully recapitulate in vivo tumor microenvironment. In this work, we demonstrate the potential of silk-alginate-based 3-D bioengineered OSCC model for S1P-targeted drug-screening towards management of OSCC. Methods: The 3-D bioengineered scaffolds based on silk fibroin from Bombyx mori cocoons and sodium alginate were fabricated using freeze-drying process. The 3-D scaffolds were characterized for various physico-chemical properties using scanning electron microscope, fourier transform infrared spectroscopy, and swelling kinetics. The cytocompatibility of the 3-D scaffolds was further assessed using MTT assay in the oral cell lines (SCC-9 and SCC-25)-seeded scaffolds. Thereafter, the gene expression of S1P metabolizing enzymes and ABCC-transporters was determined in the 3-D scaffold-based OSCC model. Finally, several compounds targeting S1P signaling along with conventional drugs 5-flurouracil and cisplatin were screened in the 3-D scaffold-based OSCC model. Results: The 3-D bioengineered silk-alginate scaffolds showed porous morphology, swelling behavior, and cytocompatibility. More specifically, cell (SCC-9 and SCC-25) seeded scaffolds demonstrated an increase in cell viability as a function of time. In addition, the 3-D scaffold-based OSCC model showed deregulated expression of the S1P metabolizing enzymes and increased expression of ABCC transporters, stemness markers and epithelial-mesenchymal transition markers. Exposure of 3-D tumor model to combination of FTY720, a functional S1P receptor antagonist and standard chemotherapeutic agents showed additive effect. Conclusions: The study demonstrated successful generation of 3-D bioengineered silk-alginate tumor model of OSCC which recapitulated properties of solid tumor and further showed potential of S1P receptor antagonist, FTY720 for management of OSCC. Citation Format: Ashok Kumar, Jahnavi Sharma, Jaya Thilakan, Pallavi Kulkarni, Vikas Gupta, Ganakalyan Behera, Neha Arya. Development of a silk-alginate-based 3-dimensional tumor model to screen anti-cancer drugs: Targeting S1P signaling for oral cancer therapy [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2025 Oct 22-26; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2025;24(10 Suppl):Abstract nr B088.