Hypoxia-inducible factor–dependent breast cancer–mesenchymal stem cell bidirectional signaling promotes metastasis
Pallavi Chaturvedi, … , Andre Levchenko, Gregg L. Semenza
Pallavi Chaturvedi, … , Andre Levchenko, Gregg L. Semenza
Published December 17, 2012
Citation Information: J Clin Invest. 2013;123(1):189-205. https://doi.org/10.1172/JCI64993.
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Research Article

Hypoxia-inducible factor–dependent breast cancer–mesenchymal stem cell bidirectional signaling promotes metastasis

Abstract

Metastasis involves critical interactions between cancer and stromal cells. Intratumoral hypoxia promotes metastasis through activation of hypoxia-inducible factors (HIFs). We demonstrate that HIFs mediate paracrine signaling between breast cancer cells (BCCs) and mesenchymal stem cells (MSCs) to promote metastasis. In a mouse orthotopic implantation model, MSCs were recruited to primary breast tumors and promoted BCC metastasis to LNs and lungs in a HIF-dependent manner. Coculture of MSCs with BCCs augmented HIF activity in BCCs. Additionally, coculture induced expression of the chemokine CXCL10 in MSCs and the cognate receptor CXCR3 in BCCs, which was augmented by hypoxia. CXCR3 expression was blocked in cocultures treated with neutralizing antibody against CXCL10. Conversely, CXCL10 expression was blocked in MSCs cocultured with BCCs that did not express CXCR3 or HIFs. MSC coculture did not enhance the metastasis of HIF-deficient BCCs. BCCs and MSCs expressed placental growth factor (PGF) and its cognate receptor VEGFR1, respectively, in a HIF-dependent manner, and CXCL10 expression by MSCs was dependent on PGF expression by BCCs. PGF promoted metastasis of BCCs and also facilitated homing of MSCs to tumors. Thus, HIFs mediate complex and bidirectional paracrine signaling between BCCs and MSCs that stimulates breast cancer metastasis.

Authors

Pallavi Chaturvedi, Daniele M. Gilkes, Carmen Chak Lui Wong, Kshitiz, Weibo Luo, Huafeng Zhang, Hong Wei, Naoharu Takano, Luana Schito, Andre Levchenko, Gregg L. Semenza

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Figure 2

HIFs mediate coculture- and hypoxia-induced CXCL10, CXCR3, CCL5, and CCR5 expression.

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HIFs mediate coculture- and hypoxia-induced CXCL10, CXCR3, CCL5, and CCR...
(A–D) BCCs, MSCs, or BCCs+MSCs were cultured at 20% or 1% O2 for 48 hours, and CXCL10 (A), CXCR3 (B), CCL5 (C), and CCR5 (D) mRNA levels were determined by RT-qPCR (mean ± SEM; n = 3). Levels were normalized to BCCs at 20% O2. *P < 0.05 vs. 20% BCCs or 20% MSCs; #P < 0.01, ##P < 0.001 vs. 20% BCCs+MSCs, 1-way ANOVA. (E and F) EV cells, DKD cells, EV+MSCs, or DKD+MSCs were cultured at 20% or 1% O2 for 48 hours. CXCL10 and CXCR3 mRNA levels were analyzed by RT-qPCR and normalized to those in EV cells at 20% O2 (mean ± SEM; n = 3). *P < 0.05 vs. 20% EV+MSCs; **P < 0.001 vs. 1% EV+MSCs, 1-way ANOVA. (G and H) GFP-expressing BCCs were cocultured with MSCs at 20% or 1% O2 for 48 hours, then subjected to FACS based on GFP fluorescence for BCCs and CD105 immunofluorescence for MSCs. CXCR3 and CXCL10 mRNA levels were determined in flow-sorted BCCs and MSCs. BCCs and MSCs cultured alone were used as controls. Levels were normalized to BCCs at 20% O2. *P < 0.01 vs. 20% MSCs or BCCs alone; #P < 0.001 vs. 1% MSCs or BCCs alone, 1-way ANOVA.