The gene Ras is mutated to remain in the active oncogenic state in one third of human cancers and as high as 90% in pancreatic cancer. We found that Ras oncogenesis is mediated differently between identical matched murine and human cells, with the latter depending upon the RalGEF pathway, suggesting a previously unappreciated key role for RalGEFs in Ras-mediated human cancer. RalGEFs activate two nearly identical G-proteins, RalA and RalB, although only the former fosters transformed and tumorigenic growth, due in part to the hypervariable C-terminus. We now propose two aims to further our understanding of the role of RalA in human cancer:
Aim 1 : Determine the function of the oncogenic C-terminus of RalA. Determining why RalA, but not RalB, can transform and promote tumor growth of human cells is important to understand Ras oncogenesis. Based on the fact that the C-terminus of RalA imparts oncogenic activity to RalB we will map the region responsible for this effect, identify the function of this region, and test if this function is required for tumorigenic growth of human pancreatic cancer cell lines.
Aim 2 : Elucidate the mechanism for RalA oncogenic signaling. How RalA is able to transform cells is unclear. We propose to attack this question at the two extreme ends of oncogenic Ral signaling. First, using loss-of-function analysis we will identify the effectors of RalA required for transformation, and test if the same effectors are also needed for tumorigenic growth of human pancreatic cancer cell lines. Second, as activation of RalA will eventually alter transcription of specific genes, we will identify a RalA oncogenic signature by comparing the transcription profile of identical cells in which RalA versus RalB is activated or repressed. The RalA oncogenic signature will be used to identify downstream pathways and specific genes associated with RalA oncogenesis. This signature will also be used to identify activation of RalA in cancers, ultimately for diagnosing cancer subtypes that rely on RalA. Completion of these aims at the basic cancer research level will better our understanding of RalA activation and signaling in cancer, and at the clinical level, provide a basis for targeting this in human cancers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA094184-09
Application #
7841769
Study Section
Cancer Molecular Pathobiology Study Section (CAMP)
Program Officer
Watson, Joanna M
Project Start
2002-02-01
Project End
2012-05-31
Budget Start
2010-06-01
Budget End
2011-05-31
Support Year
9
Fiscal Year
2010
Total Cost
$258,320
Indirect Cost
Name
Duke University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Kashatus, Jennifer A; Nascimento, Aldo; Myers, Lindsey J et al. (2015) Erk2 phosphorylation of Drp1 promotes mitochondrial fission and MAPK-driven tumor growth. Mol Cell 57:537-51
Brady, Donita C; Crowe, Matthew S; Turski, Michelle L et al. (2014) Copper is required for oncogenic BRAF signalling and tumorigenesis. Nature 509:492-6
Yang, Shuqun; Counter, Christopher M (2013) Cell cycle regulated phosphorylation of the telomere-associated protein TIN2. PLoS One 8:e71697
Kashatus, David F; Lim, Kian-Huat; Brady, Donita C et al. (2011) RALA and RALBP1 regulate mitochondrial fission at mitosis. Nat Cell Biol 13:1108-15
Issaq, Sameer H; Lim, Kian-Huat; Counter, Christopher M (2010) Sec5 and Exo84 foster oncogenic ras-mediated tumorigenesis. Mol Cancer Res 8:223-31
Lim, Kian-Huat; Brady, Donita C; Kashatus, David F et al. (2010) Aurora-A phosphorylates, activates, and relocalizes the small GTPase RalA. Mol Cell Biol 30:508-23
Zipfel, P A; Brady, D C; Kashatus, D F et al. (2010) Ral activation promotes melanomagenesis. Oncogene 29:4859-64
Naini, Sarasija; Etheridge, Katherine T; Adam, Stacey J et al. (2008) Defining the cooperative genetic changes that temporally drive alveolar rhabdomyosarcoma. Cancer Res 68:9583-8
Adam, Stacey J; Counter, Christopher M (2008) A method to generate genetically defined tumors in pigs. Methods Enzymol 439:39-51
Lim, Kian-Huat; Ancrile, Brooke B; Kashatus, David F et al. (2008) Tumour maintenance is mediated by eNOS. Nature 452:646-9

Showing the most recent 10 out of 24 publications