Two distinct mTORC2-dependent pathways converge on Rac1 to drive breast cancer metastasis

• Published in: Breast cancer research : BCR 2017-06, Vol.19 (1), p.74-74, Article 74
• Main Authors: Morrison Joly, Meghan, Williams, Michelle M, Hicks, Donna J, Jones, Bayley, Sanchez, Violeta, Young, Christian D, Sarbassov, Dos D, Muller, William J, Brantley-Sieders, Dana, Cook, Rebecca S
• Format: Article
• Language: English
• Subjects: Akt; AKT protein; Analysis; Animal models; Animals; Breast cancer; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Breast Neoplasms - mortality; Breast Neoplasms - pathology; Cancer metastasis; Cell adhesion & migration; Cell cycle; Cell Line, Tumor; Cell Movement - genetics; Cell survival; Development and progression; Disease Models, Animal; Epidermal growth factor; ErbB-2 protein; Female; Gene Amplification; Genetic aspects; Genetic engineering; Guanine; Guanine nucleotide exchange factor; Guanosine; Guanosine triphosphate; Heterografts; Humans; Kinases; Mechanistic Target of Rapamycin Complex 2 - metabolism; Metastases; Metastasis; Mice; Mice, Transgenic; Motility; mTOR; Neoplasm Metastasis; Neoplasm Staging; Physiological aspects; Prognosis; Protein kinase C; Protein kinases; Proteins; Proto-Oncogene Proteins c-akt - metabolism; Rac; rac1 GTP-Binding Protein - metabolism; Rac1 protein; Rapamycin-Insensitive Companion of mTOR Protein - genetics; Rapamycin-Insensitive Companion of mTOR Protein - metabolism; Receptor, ErbB-2 - genetics; Receptor, ErbB-2 - metabolism; rho Guanine Nucleotide Dissociation Inhibitor beta - genetics; rho Guanine Nucleotide Dissociation Inhibitor beta - metabolism; Rictor; Signal Transduction; Stem cells; Survival; Tiam1 protein; TOR protein; Tumor cell lines; Tumor cells; Tumors
• ISSN: 1465-542X; 1465-5411; 1465-542X
• DOI: 10.1186/s13058-017-0868-8

The importance of the mTOR complex 2 (mTORC2) signaling complex in tumor progression is becoming increasingly recognized. HER2-amplified breast cancers use Rictor/mTORC2 signaling to drive tumor formation, tumor cell survival and resistance to human epidermal growth factor receptor 2 (HER2)-targeted therapy. Cell motility, a key step in the metastatic process, can be activated by mTORC2 in luminal and triple negative breast cancer cell lines, but its role in promoting metastases from HER2-amplified breast cancers is not yet clear. Because Rictor is an obligate cofactor of mTORC2, we genetically engineered Rictor ablation or overexpression in mouse and human HER2-amplified breast cancer models for modulation of mTORC2 activity. Signaling through mTORC2-dependent pathways was also manipulated using pharmacological inhibitors of mTOR, Akt, and Rac. Signaling was assessed by western analysis and biochemical pull-down assays specific for Rac-GTP and for active Rac guanine nucleotide exchange factors (GEFs). Metastases were assessed from spontaneous tumors and from intravenously delivered tumor cells. Motility and invasion of cells was assessed using Matrigel-coated transwell assays. We found that Rictor ablation potently impaired, while Rictor overexpression increased, metastasis in spontaneous and intravenously seeded models of HER2-overexpressing breast cancers. Additionally, migration and invasion of HER2-amplified human breast cancer cells was diminished in the absence of Rictor, or upon pharmacological mTOR kinase inhibition. Active Rac1 was required for Rictor-dependent invasion and motility, which rescued invasion/motility in Rictor depleted cells. Rictor/mTORC2-dependent dampening of the endogenous Rac1 inhibitor RhoGDI2, a factor that correlated directly with increased overall survival in HER2-amplified breast cancer patients, promoted Rac1 activity and tumor cell invasion/migration. The mTORC2 substrate Akt did not affect RhoGDI2 dampening, but partially increased Rac1 activity through the Rac-GEF Tiam1, thus partially rescuing cell invasion/motility. The mTORC2 effector protein kinase C (PKC)α did rescue Rictor-mediated RhoGDI2 downregulation, partially rescuing Rac-guanosine triphosphate (GTP) and migration/motility. These findings suggest that mTORC2 uses two coordinated pathways to activate cell invasion/motility, both of which converge on Rac1. Akt signaling activates Rac1 through the Rac-GEF Tiam1, while PKC signaling dampens expression of the e