Angiogenesis, the process that new blood vessels form from existing vessels, is a critical process in development and response to environment and disease. Alterations in vascular development and angiogenesis play important roles in many diverse disease including diabetes, coronary artery disease, stroke, and cancer. More complete understanding of the mechanisms leading to appropriately controlled angiogenesis may lead to new approaches for prevention and/or treatment of these common debilitating diseases. Capillary tubules form from endothelial cells by a process called "cell-hollowing" tubulogenesis, which involves formation and fusion of intracellular vesicles to form a large intracellular vacuole that eventually becomes the extracellular lumen of the tubule. CLIC proteins are a family of chloride channel proteins which largely reside in intracellular membranes. Chloride permeability has long been recognized to play a role in intracellular membrane traffic but the specific proteins responsible for individual chloride conductance of intracellular membranes is still largely unknown. Recently, a CLIC protein has been shown to be essential for a specific cell-hollowing tubulogenic process in C. elegans and the family member CLIC4 has been implicated in tubulogenesis in vitro of mammalian endothelial cells. We have recently generated and characterized mice in which the gene for CLIC4 has been disrupted. Preliminary data from these mice provide startling and exciting new evidence that CLIC4 is indeed involved in angiogenesis. Our hypothesis is that CLIC4 plays a critical role in angiogenesis, perhaps by providing the chloride conductance necessary for the orderly membrane traffic and membrane fusion steps involved in cell-hollowing tubulogenesis of endothelial cells. We propose to: 1) Study CLIC4 (-/-) mice for abnormalities in vascular development and response to angiogenic stimuli;2) Identify sequences and motifs within CLIC4 that are necessary for its function in cell-hollowing tubulogenesis;and 3) Investigate transport properties of membranes derived from cells undergoing tubulogenesis for an activity attributable to CLIC4 and explore how that effects tubulogenesis. The proposal thus is a hypothesis-driven basis science project that uses novel approaches and reagents to address a recently-identified key step in angiogenesis about which little is currently known mechanistically. The results will have clear clinical relevance to the many diseases in which angiogenesis plays a part.

Public Health Relevance

Growth and regulation of new blood vessels is important in normal development and critical in several important diseases such as cancer, heart disease and stroke. This proposal focuses on newly discovered key role for an intracellular chloride channel in development of new blood vessels. The results of the proposed studies will provide new insight into these common, important disease processes and may open new avenues for prevention and treatment.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL092131-05
Application #
8282835
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Gao, Yunling
Project Start
2009-07-17
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2013-05-31
Support Year
5
Fiscal Year
2012
Total Cost
$382,332
Indirect Cost
$127,444
Name
Saint Louis University
Department
None
Type
Schools of Medicine
DUNS #
050220722
City
Saint Louis
State
MO
Country
United States
Zip Code
63103
Wojciak-Stothard, Beata; Abdul-Salam, Vahitha B; Lao, Ka Hou et al. (2014) Aberrant chloride intracellular channel 4 expression contributes to endothelial dysfunction in pulmonary arterial hypertension. Circulation 129:1770-80
Edwards, John C; Bruno, Jonathan; Key, Phillip et al. (2014) Absence of chloride intracellular channel 4 (CLIC4) predisposes to acute kidney injury but has minimal impact on recovery. BMC Nephrol 15:54
Edwards, John C; Kahl, Christina R (2010) Chloride channels of intracellular membranes. FEBS Lett 584:2102-11