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Tumor-derived osteopontin drives the resident fibroblast to myofibroblast differentiation through Twist1 to promote breast cancer progression

Abstract

Tumor-stroma interactions are important determinants for the disease course in cancer. While stromal influence has been known to often play a tumor-promoting role, incomplete mechanistic insight into this phenomenon has prevented its therapeutic targeting. Stromal fibroblasts can be activated by tumor cells to differentiate into cancer-associated fibroblasts (CAFs), that exhibit the traits of myofibroblasts, and in turn, they increase cancer aggressiveness. Here, we report the crosstalk between the cancer cells and stromal fibroblasts that leads to tumor progression. The process is initiated by secretion of a chemokine like protein, osteopontin (OPN) from the cancer cells that differentiates the fibroblasts to myofibroblasts. Tumor-derived OPN achieves this transition by engaging CD44 and αvβ3 integrins on the fibroblast surface, which mediates signaling via Akt and ERK to induce Twist1-dependent gene expression. The OPN-driven CAFs then secrete CXCL12, which in turn triggers epithelial to mesenchymal transition (EMT) in the tumor cells. OPN, produced by the cancer cells, and CXCL12, secreted by activated fibroblasts, are necessary and sufficient to perpetuate the crosstalk. Knocking out OPN in carcinogen-induced mammary tumors or knocking down OPN in cancer cells and fibroblast co-implanted xenografts abrogates myofibroblast differentiation, Twist1, and CXCL12 expression. OPN expression is correlated with CAF-specific gene signature as shown by breast tumor tissue microarray consisting of 100 patient specimens. Bioinformatics analyses have confirmed that the expression of OPN is significantly correlated with the expression of myofibroblast-specific markers as demonstrated in human breast carcinoma dataset of 2509 patients. Our findings describe OPN and CXCL12 act as compelling targets to curb the tumor-promoting features of the stromal components and further suggested that OPN-regulated CXCL12 network might act as potential therapeutic target for the management of CAF-mediated breast cancer progression.

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Fig. 1: Breast cancer cell-derived OPN increases migratory ability and upregulates myofibroblast-specific marker expressions in fibroblasts.
Fig. 2: OPN is sufficient to reprogram normal fibroblasts into myofibroblasts.
Fig. 3: Tumor-derived OPN induces myofibroblast differentiation in a co-implanted xenograft model.
Fig. 4: OPN-educated fibroblasts induce cancer progression via CXCL12 (SDF-1) expression.
Fig. 5: Correlation of OPN with expression of myofibroblast-specific markers in human breast cancer tissues.
Fig. 6: Molecular mechanisms underlying OPN-induced CAF trans-differentiation.

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Acknowledgements

This work was primarily supported by National Centre for Cell Science, Pune, India (to GCK). RB, RN, and DT were supported by Council of Scientific and Industrial Research (CSIR), Govt. of India. VPG is supported by Science and Engineering Research Board (SERB), Department of Science and Technology (DST), Govt. of India. This research was supported in part by the Intramural Research Program of the NIH, National Institute of Ageing.

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RB designed and performed most of the experiments. RN, TVS, AB, VPG, DT, and DK performed some of the experiments. GK analyzed patient datasets. AM, SG, and TP helped in acquiring patient samples and clinical data analysis. GFW was involved in writing and critically reading the manuscript. GCK supervised and designed the whole study and critically assessed the manuscript.

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Correspondence to Gopal C. Kundu.

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Butti, R., Nimma, R., Kundu, G. et al. Tumor-derived osteopontin drives the resident fibroblast to myofibroblast differentiation through Twist1 to promote breast cancer progression. Oncogene 40, 2002–2017 (2021). https://doi.org/10.1038/s41388-021-01663-2

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