Malignant gliomas are one of the most aggressive and deadly forms of cancer and can affect any age group. In glioblastoma multiforme (GBM), infiltration of primary tumor cells into the normal tissue and dispersal throughout the brain is a central challenge to successful treatment that remains unmet. Patients with malignant gliomas respond poorly to the standard therapeutic regimen of radiotherapy and chemotherapy that follow tumor resection and have only a 16-month median survival. It is therefore imperative to identify new approaches to specifically attack GBM cell survival, proliferation and invasion. The cellular mechanisms driving GBM-mediated migration and invasion are not fully understood. RSK2 (p90 ribosomal S6 kinase 2) is a kinase that regulates proliferation and adhesion and can promote metastasis. We find that RSK2 is significantly upregulated in vivo in human GBM patient tumors, and that high RSK2 expression significantly correlates with advanced tumor stage and poor patient survival. We demonstrate that active RSK2 regulates GBM adhesion and is essential for cell motility and invasion of patient-derived GBM neurospheres. Importantly, inhibition of RSK2 by either RSK inhibitors or shRNA silencing impairs invasion and combining RSK2 inhibitors with temozolomide improves efficacy in vitro. We further show that the effects of RSK2 on GBM invasion are mediated in part through activation of Rho GTPases. Rho family of GTPases are key regulators of cell polarity, migration and invasion. Here, we identify Rho A, B and C as downstream effectors of (RSK2) in GBM cell migration and invasion. Here, we identified a unique signaling pathway in which RSK2 confers invasiveness through leukemia-associated RhoGEF (LARG)-dependent Rho GTPase activation. RSK2 directly interacts with LARG and nucleotide-bound Rho isoforms, but not Rac1 and Cdc42. Our data support a fundamental importance of residue of Thr577 to the invasive signaling of RSK2. The invasive signaling of RSK2 appears to depend on RhoA and B, but not RhoC. RSK2 activates RhoA through activation of LARG by phosphorylation at Ser1288. Intriguingly, LARG activity is only required for RSK2-T577E, but not RSK2-Y707A-mediated RhoA activation and cell invasion. This further supports a pivotal role of Thr577 in invasive RSK2 signaling. These results establish a unique RSK2-dependent LARG-Rho signaling cascade as an uncompensated factor key to GBM cell migration and invasion. Together, our data provide strong evidence that RSK inhibitors could enhance the effectiveness of existing GBM treatment, and support RSK2 targeting as a promising approach for novel GBM therapy.

Citation Format: Geng-Xian Shi, Ling Jin, Michelle L. Matter, Santosh Kesari, Joe W. Ramos. RSK2 provokes invasive signaling in glioblastoma through LARG-dependent activation of Rho GTPases [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1363. doi:10.1158/1538-7445.AM2017-1363