Abstract A41: GPRC5A acts as a potent oncogene in pancreatic cancer
Authors: Honglei Zhou, Aristeidis Telonis, Yi Jing, Masaya Jimbo, Fernando Blanco, Eric Londin, Jonathan Brody, Isidore Rigoutsos
Journal: Cancer Research (2016)

Abstract Background: GPRC5A is an orphan G-protein coupled receptor acting as an oncogene in some cancers and as a tumor suppressor in other. Its role in pancreatic cancer has not been studied systematically. ELAV-like protein 1, also known as “HuR,” is an RNA binding protein that is encoded by the ELAVL1 gene. HuR post-transcriptionally regulates multiple genes, plays key roles in human malignancies, and is known to mediate the efficacy of gemcitabine. Our preliminary data indicated that, in pancreatic cells, GPRC5A transcription responded to gemcitabine treatment and that its mRNA likely interacted with HuR. Methods: We immunohistochemically analyzed GPRC5A protein expression in primary and metastatic pancreatic cancer samples. We examined the effects of GPRC5A on pancreatic cells’ ability to proliferate, migrate, and invade, and how gemcitabine modulates this ability. We also studied the interaction between GPRC5A and HuR and examined how it is affected by gemcitabine. Lastly, we used microarray expression analyses to identify the genes and pathways linked to GPRC5A in pancreatic cancer cells. Results: We analyzed public mRNA expression data from 675 human cancer cell lines and 10,609 samples from The Cancer Genome Atlas (TCGA): GPRC5A mRNA abundance in pancreatic cancer was the highest (cell lines) or second highest (TCGA) among all tissues and cancer types. We analyzed an independent set of 252 pancreatic normal and cancer samples: we found GPRC5A mRNA to be up-regulated in primary tumor samples compared to normal pancreas, and even further up-regulated in metastatic pancreatic cancer. We immunostained 208 cores (103 primary and metastatic pancreatic cancer samples): we found GPRC5A protein to be barely expressed in normal pancreatic ductal cells and highly expressed in both primary and metastatic pancreatic ductal cancer cells. We carried out in vitro studies of multiple pancreatic cancer cell lines: we found that increasing GPRC5A protein levels promoted pancreatic cancer cell growth and migration whereas siRNA-mediated knockdown of GPRC5A impaired proliferation and invasion. We found that gemcitabine treatment enhanced GPRC5A expression in pancreatic cancer cells. Conversely, siRNA-mediated knockdown of GPRC5A sensitized the cells to gemcitabine. We showed that HuR/ELAVL1 up-regulated GPRC5A protein in the first 18 hours following gemcitabine treatment by stabilizing GPRC5A’s mRNA through binding to at least on site in GPRC5A’s 3´UTR. Microarray analysis showed that GPRC5A overexpression impacts several signaling networks. Conclusions: Our results indicate that GPRC5A acts as an oncogene in pancreatic cancer. Unexpectedly, gemcitabine was found to increase GPRC5A's mRNA and protein levels. We showed that this increase is mediated by HuR, a known enabler of gemcitabine efficacy, through a direct interaction between HuR and GPRC5A’s mRNA. It appears that the sensitivity of pancreatic cancer cells to gemcitabine can be augmented through down-regulation of GPRC5A. Citation Format: Honglei Zhou, Aristeidis Telonis, Yi Jing, Masaya Jimbo, Fernando Blanco, Eric Londin, Jonathan Brody, Isidore Rigoutsos.{Authors}. GPRC5A acts as a potent oncogene in pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A41.