Abstract B036: Platelets promote in vitro osteosarcoma cell migration and in vivo metastatic lung seeding

Abstract

Abstract Osteosarcoma (OS) is the most common primary bone cancer worldwide where it mainly affects adolescents. Metastatic OS has a 20% survival rate and treatment options remain poor, thus understanding mechanisms that contribute to OS metastasis is critical for improved patient outcomes. Platelets, commonly recognized for their role in hemostasis, have been shown to promote metastasis in several cancer models. However, most previous studies investigating platelet contributions to metastasis have focused on carcinomas while their role in osteosarcoma remains poorly understood. Previous studies by our lab have shown that platelet interactions with breast cancer cells resulted in increased migration in vitro and increased lung metastases in vivo. Thus, we hypothesized that OS cell migration will be enhanced in the presence of platelets, and that platelet depletion in vivo will inhibit OS lung seeding. Cell suspensions of SaOs2 and MG63.3 cells in serum free media were fluorescently stained and seeded, in triplicates, in the top chamber of an 8um pore Fluoroblock migration assay wells. In the bottom chamber, media and either Tyrode’s buffer or 1.5x106 isolated human platelets were added. The presence of platelets resulted in a 3.5- and 4.6-fold increase of migrated MG63.3 and SaOs2 cells respectively. For in vivo analysis, cohorts of eight-week-old, male Nod Scid-Gamma (NSG), Nod Scid, and Nude mice were randomly assigned to either the control or platelet depletion group. Mice were depleted of platelets by administration of Anti-Platelet glycoprotein Ib alpha (GPIbα) antibody and, 5x105 GFP transduced MG63.3 cells were injected via tail vein. 24 hrs later, the mice were euthanized, and lungs were collected and microscopically imaged. Platelet depletion in NSG mice resulted in a 48% reduction in MG63.3 lung seeding compared to the isotype control recipient mice. This result was more dramatic in the less immunocompromised Nude and Nod Scid mice with a 61% and 87% reduction of MG63.3 lung seeding in the platelet depleted mice respectively. Finally, RNAseq and was performed on seven OS cell lines co-cultured with isolated mouse platelets, and alterations in gene expression were compared to control cells. Gene Enrichment Set Analysis (GSEA) of the data indicated that platelet co-incubation significantly upregulated pathways related to inflammatory responses, Tumor Necrosis Factor (TNF)-a signaling, and Interleukin (IL)-6 signaling. These results suggest that platelet secreted factors facilitate OS cell migration in vitro while an immune dependent mechanism is involved in the role of platelets in the in vivo lung seeding of OS cells. Citation Format: Olivia K. Travis, Ruishan Wang, Myriam Labelle. Platelets promote in vitro osteosarcoma cell migration and in vivo metastatic lung seeding [abstract]. In: Proceedings of the AACR Special Conference: Cancer Metastasis; 2022 Nov 14-17; Portland, OR. Philadelphia (PA): AACR; Cancer Res 2022;83(2 Suppl_2):Abstract nr B036.