The high frequency (95%) of KRAS mutations in pancreatic ductal adenocarcinomas (PDAC), together with strong evidence for the KRAS dependency of PDAC growth, makes the K-Ras oncoprotein an attractive therapeutic target. However, to date the development of direct inhibitors of K-Ras has not been successful, with most efforts focused on targeting components of K-Ras downstream effector signaling. While the pharmaceutical industry has centered on inhibitors of the Raf or PI3K effector signaling pathways, there is substantial evidence that K-Ras-dependent cancer growth must rely on the activities of other effector pathways. Of those pathways, the Ral guanine nucleotide exchange factor RalGEF) effector pathway, leading to activation of the Ras-like (Ral) small GTPases is well-validated to drive PDAC growth. Interestingly, while the two Ral isoforms share significant sequence identity (82%) as well as the same set of effectors, our studies found that they have strikingly different roles in PDAC growth. While RalA but not RalB is necessary for PDAC anchorage-independent growth in vitro and tumorigenic growth in vivo, RalB is the critical driver of PDAC invasion in vitro and metastasis in vivo. The mechanistic basis for their different biological roles in cancer remains to be elucidated. We hypothesize that their divergent C-terminal sequences and the posttranslational modifications that they signal for result in distinct subcellular localizations that then contribute to Ral isoform-selective effector interactions that promote distinct biological consequences in PDAC. To evaluate our hypothesis, we initiated studies to determine the effector(s) that mediates RalA-dependent anchorage-independent growth and tumorigenesis in vivo, the effector(s) that mediates RalB-dependent invasion and metastasis in vivo; and the role of the C-terminal hypervariable domain in causing distinct RalA- and RalB-dependent phenotypes and effector utilization. We found that both RalA and RalB subcellular localization is dependent on the C-terminal CAAX tetrapeptide signaled modifications by Rce1 and ICMT. We also determined that RalB preferentially activated mTORC1 to regulate pancreatic tumor cell Matrigel migration, through the Sec5 effector and regulation of exocyst function.
Citation Format: Leanna R. Gentry, Timothy D. Martin, David J. Reiner, Channing J. Der. Mechanistic dissection of Ral GTPase signaling in driving KRAS-dependent pancreatic cancer growth. [abstract]. In: Proceedings of the AACR Special Conference on RAS Oncogenes: From Biology to Therapy; Feb 24-27, 2014; Lake Buena Vista, FL. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(12 Suppl):Abstract nr A08. doi: 10.1158/1557-3125.RASONC14-A08