G protein-coupled receptors (GPCRs) are well known for their ability to convert extracellular information encoded in hormones, odorants, and peptides into rapid changes in cellular homeostasis by modulating the function of effector enzymes and channels in the cell. However, analysis of human cancer genomes indicates that some GPCR signaling pathways lead to sustained signals that in pathological settings are linked to tumor growth and metastasis. A common feature of such pathways is the activation of Rho GTPases by Dbl family Rho guanine nucleotide exchange factors (RhoGEFs). The TrioC subfamily (composed of Trio, Kalirin, and p63RhoGEF) is directly activated by heterotrimeric G?q subunits and is strongly implicated in ocular melanoma and leukemia. The P-Rex subfamily (composed of P-Rex1 and 2) is activated synergistically by direct interactions with G?? and the lipid PIP3, and is overexpressed in many breast and prostate tumors where it plays a metastatic role. Trio and P-Rex1/2 have thus emerged as important chemotherapeutic targets. The fact that the molecular and cellular mechanisms underlying regulation of these enzymes are as of yet poorly understood prevents a rational approach to the design of novel therapeutic approaches. By determining crystallographic and cryo-EM structures of members from these RhoGEF subfamilies, this proposal seeks to define molecular mechanisms of activation and to test these hypotheses through a battery of functional assays. In parallel, we extend our functional analysis into model cell systems to understand how constitutively active heterotrimeric G proteins and GPCRs promote cancer cell growth (as in ocular melanoma) and metastasis (as in breast cancer) via TrioC and P-Rex subfamily RhoGEFs. By understanding the basis for regulation of RhoGEF activity by heterotrimeric G proteins both in vitro and in relevant cellular contexts, we will accelerate discovery of new biological insights and novel therapeutic strategies that can be used to combat cancer.

Public Health Relevance

G protein-coupled receptors (GPCRs) are well known for their ability to rapidly alter cell physiology in response to extracellular stimuli, but a subset of their downstream pathways are increasingly recognized for their ability to promote sustained growth and chemotaxis and for their participation in tumor growth and metastasis. This proposal seeks to understand the molecular basis underlying the regulation of two subfamilies of GPCR- regulated Rho GTPase guanine nucleotide exchange factors that play roles in cancer-related pathways, with the long-term goal of developing novel approaches to selectively inhibit these enzymes.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA221289-02
Application #
9653982
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Knowlton, John R
Project Start
2018-03-01
Project End
2023-02-28
Budget Start
2019-03-01
Budget End
2020-02-29
Support Year
2
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Purdue University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907