Abstract PR010: Development of a pre-clinical metastatic model of human sarcoma to identify therapeutic targets

Abstract

Abstract Targeted therapies have led to significant advances in the treatment of multiple tumor types resulting in more effective and often, less toxic therapeutic options. In sarcomas, the development of targeted therapies has been met limited success. There are more than 70 sarcoma subtypes that vary in histology, clinical course and patient demographics. Despite these clear differences, clinically, sarcomas are treated similarly with variable efficacy. Patients with localized disease are treated with surgery, radiation, and often chemotherapy. Even with this aggressive multimodality treatment, 35% of patients will develop incurable metastatic disease. This highlights the need for additional therapies targeting micrometastatic disease or cells within the primary tumor with a high propensity for metastasis. Understanding the pathways driving the formation of sarcoma metastasis would allow for the development of new therapies. Targeted treatments are limited due to the heterogeneity of the disease and the paucity of pre-clinical models that accurately reflect the human disease. We have focused on creating in vivo models of sarcoma development and metastasis that can be used as the basis of further studies and to test potential therapeutic targets. Mesenchymal stem cells (MSCs) are the presumed cell of origin for sarcomas and therefore, the starting cell for our investigations. We hypothesized that by recreating key genetic events in human MSCs, we could generate sarcomas in vivo that are reflective of the human disease. RB1 and P53, tumor suppressors that are often mutated or functionally inactive in sarcomas were first targeted using CRISPR-Cas9 technology in MSCs that have been immortalized by human telomerase (hTERT). Genes that are overexpressed or amplified in The Cancer Genome Atlas (TCGA) data were identified and a library of potential oncogenes was generated. This library was then added to RB1-/-P53+/- cells through lentiviral transduction. Targeting of key tumor suppressors and adding oncogenic drivers resulted in the formation of high-grade human sarcomas subcutaneously. We then sought to investigate the ability of these cells to metastasize. Injection of cells intramuscularly (into the thigh) in immunocompromised mice resulted in the formation of spontaneous lung metastasis without clear evidence of disease in other organs. This pattern clinically reflects that of human disease. Comparing oncogenic genes from metastatic outgrowths to primary tumors identified KLF4, DDIT3, JUN, and KRAS as being enriched in metastatic cells. This system allows for reproducible and robust genetic manipulation of tumor cells to characterize key drivers of metastatic growth. This will result in the identification and validation of new therapeutic targets to treat or prevent metastatic disease in our patients. Citation Format: Janai R. Carr-Ascher. Development of a pre-clinical metastatic model of human sarcoma to identify therapeutic targets [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr PR010.