Angiogenesis, the production of new blood vessels from existing vasculature, is required for normal development, wound repair, and vascular homeostasis. Neuropilin is an essential mammalian cell surface receptor for one of the most potent pro-angiogenic molecules, vascular endothelial growth factor (VEGF), and functions in synchrony with VEGFR receptor tyrosine kinases. Neuropilin is of particular medical importance because it is critical to the pathology of VEGF dependent hyper-vascularization in tumor angiogenesis. We propose a series of parallel and complementary structural and functional studies to understand the physical interactions and receptor coupling at the heart of the VEGF signaling complex. Unlike many other kinase systems, VEGF binding to VEGFR in the absence of neuropilin is insufficient to induce angiogenesis. We seek to first understand the molecular mechanism(s) underlying neuropilin's unique role in VEGF dependant angiogenesis.
In Aim 1, we will determine the basis for competitive binding of ligands to neuropilin, and the role that this competition plays in regulating angiogenesis.
In Aim 2, we will explore the structural and conformational changes that neuropilin undergoes in its transition from inactive monomer to activated dimer. More globally, we seek to define the physical basis for the coordinated action of VEGF, neuropilin, and VEGFR.
Aim 3 is focused on utilizing multiple structural and biochemical tools coupled with endothelial cell signaling assays to determine the physical interactions and conformational changes necessary to couple extracellular ligand binding to receptor activation. Taken together, these studies will provide a molecular understanding of the physical mechanisms underlying the critical first step connecting VEGF ligand binding to receptor activation.

Public Health Relevance

Neuropilin is an essential receptor needed for the formation of new blood vessels. New blood vessels are critical during normal development and wound repair but also provide a tumor with critical nutrients for its growth. Understanding how neuropilin works in the formation of new blood vessels will help us understand how this pathway normally functions and how we can block its activity in tumors and related diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM094155-04
Application #
8502185
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Dunsmore, Sarah
Project Start
2010-09-06
Project End
2015-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
4
Fiscal Year
2013
Total Cost
$248,982
Indirect Cost
$81,318
Name
University of Kentucky
Department
Biochemistry
Type
Schools of Medicine
DUNS #
939017877
City
Lexington
State
KY
Country
United States
Zip Code
40506
Goel, Hira Lal; Pursell, Bryan; Shultz, Leonard D et al. (2016) P-Rex1 Promotes Resistance to VEGF/VEGFR-Targeted Therapy in Prostate Cancer. Cell Rep 14:2193-208
Thacker, Bryan E; Seamen, Emylie; Lawrence, Roger et al. (2016) Expanding the 3-O-Sulfate Proteome--Enhanced Binding of Neuropilin-1 to 3-O-Sulfated Heparan Sulfate Modulates Its Activity. ACS Chem Biol 11:971-80
Guo, Hou-Fu; Vander Kooi, Craig W (2015) Neuropilin Functions as an Essential Cell Surface Receptor. J Biol Chem 290:29120-6
Parker, Matthew W; Linkugel, Andrew D; Goel, Hira Lal et al. (2015) Structural basis for VEGF-C binding to neuropilin-2 and sequestration by a soluble splice form. Structure 23:677-87
Li, Xiaobo; Parker, Matthew W; Vander Kooi, Craig W (2014) Control of cellular motility by neuropilin-mediated physical interactions. Biomol Concepts 5:157-66
Wu, Tao; Kooi, Craig Vander; Shah, Pritom et al. (2014) Integrin-mediated cell surface recruitment of autotaxin promotes persistent directional cell migration. FASEB J 28:861-70
Parker, Matthew W; Vander Kooi, Craig W (2014) Microplate-based screening for small molecule inhibitors of neuropilin-2/vascular endothelial growth factor-C interactions. Anal Biochem 453:4-6
Parker, Matthew W; Linkugel, Andrew D; Vander Kooi, Craig W (2013) Effect of C-terminal sequence on competitive semaphorin binding to neuropilin-1. J Mol Biol 425:4405-14
Guo, Hou-Fu; Li, Xiaobo; Parker, Matthew W et al. (2013) Mechanistic basis for the potent anti-angiogenic activity of semaphorin 3F. Biochemistry 52:7551-8
Goel, Hira Lal; Pursell, Bryan; Chang, Cheng et al. (2013) GLI1 regulates a novel neuropilin-2/α6β1 integrin based autocrine pathway that contributes to breast cancer initiation. EMBO Mol Med 5:488-508

Showing the most recent 10 out of 15 publications