Abstract 1394: The CXCL10-CXCR3 axis as Jekyll and Hyde: A regulator of metastasis and immune response in osteosarcoma
Authors: Gyau Benjamin B. , Chen Xiang , Wang Junyan , Clement Margaret A. , Major Angela M. , Xu Jun , Hicks M. John , Man Tsz-Kwong 
Published: March 2024

Abstract
The CXCL10-CXCR3 axis is known to promote tumor growth and metastasis via autocrine signaling, while it can also elicit anti-tumor responses by paracrine signaling. However, its roles are still elusive in osteosarcoma (OS), the most common malignant bone tumor in children. In this study, we utilized in vitro assays, mouse models, and gene expression analysis to characterize the roles of the CXCL10-CXCR3 axis in OS. To understand the autocrine signaling, we performed in vitro phenotypic assays on three OS cell lines with and without CXCL10. The results showed that the chemokine increased AKT phosphorylation and tumor cell migration. Using a CRISPR-based CXCR3 deletion mutant, we demonstrated that the lack of the receptor inhibited OS tumor growth and pulmonary metastasis in an orthotopic xenograft mouse model. The results indicate that the CXCL10-CXCR3 axis is sufficient and necessary to promote aggressive phenotypes in OS via autocrine signaling. Next, using gene expression datasets from two cohorts of OS patients, we showed that high expression of CXCL10 or its cognate receptor CXCR3 was associated with a better prognosis. Since CXCL10 is known to recruit CXCR3+ immune cells to fight against cancer, we further found that the expression of the T cell marker CD3D was associated with a better prognosis. These results suggest that the chemokine may also play a protective role in osteosarcoma by recruiting anti-tumor immune cells via paracrine signaling. Lastly, we have previously reported that a high circulating level of CXCL10 correlates with a poor prognosis in osteosarcoma patients. In this study, we demonstrated that increasing the circulating CXCL10 level in immunodeficient orthotopic xenograft mouse models of OS did not significantly promote the development of pulmonary metastases, suggesting circulating CXCL10 may exert its chemotactic effect mainly on immune cells, not on tumor cells. Taken together, we propose a model that tumor expression of CXCL10 promotes osteosarcoma growth and metastasis development via autocrine signaling. The increase of the local level of CXCL10 attracts CXCR3+ immune cells to the tumor site and promotes anti-tumor function via paracrine signaling. Metastatic cells colonize the lungs and cause tissue damage and inflammation, which increases the production of CXCL10 in the circulation. The higher circulating CXCL10 recruits T cells away from the primary tumor site and indicates the occurrence of metastasis and, hence, correlates with a worse outcome in patients. Our research highlights the importance of understanding the opposite effects of CXCL10 on tumor and immune cells as well as in the tumor microenvironment and circulation. Identification of the mechanistic differences in the CXCL10-CXCR3 signaling between tumor and immune cells will provide a novel therapeutic approach against tumor cells while promoting anti-tumor immune response in OS.
Citation Format: Benjamin B. Gyau, Xiang Chen, Junyan Wang, Margaret A. Clement, Angela M. Major, Jun Xu, M. John Hicks, Tsz-Kwong Man. The CXCL10-CXCR3 axis as Jekyll and Hyde: A regulator of metastasis and immune response in osteosarcoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1394.