Brain arteriovenous (AV) malformations (BAVMs) can cause life-threatening strokes and have limited treatment options. The goal for this project is to elucidate the cellular and molecular mechanisms underlying BAVM pathogenesis to identify novel candidates as therapeutic targets to ameliorate this disease. AVMs are characterized by abnormal AV shunts that displace intervening capillaries. We propose a cross-disciplinary approach, fusing cutting-edge mouse genetics and imaging technologies, to test our hypothesis that Notch mutations can reprogram AV identity and alter AV differential nitric oxide (NO) signaling, and thus endothelial dysfunction, to elicit BAVMs. Notch receptors and ligands are expressed in arteries but not veins. Notch signaling promotes arterial at the expense of venous differentiation by enhancing arterial and suppressing venous molecular markers. We have reported that endothelial expression of a constitutively active Notch4 mutation (Notch4*) elicits BAVMs in mice. Notch4* reprograms veins to gain arterial and lose venous molecular identity, and correcting the causal Notch4* leads to normalization of established BAVMs. We have built a custom two-photon microscope, optimal for structural and hemodynamic imaging of cerebral vasculature in live mice. We can thus obtain 5D data (3D plus blood velocity over time) through a cranial window to reveal the process of BAVM formation in mice. Built on our strong background and preliminary data, we propose:
Aim 1 - Determine the effect of endothelial Notch4* on venous endothelial dysfunction and BAVM formation. We will test our hypothesis that Notch4* upregulates NO levels in the veins, alters venous endothelial response to blood flow, and thus permits AVM formation. We will examine the effect of Notch4* on venous NO signaling, endothelial response to blood flow, and flow mediated BAVM formation;
Aim 2 - Determine the effect of endothelial Notch deficiency on arterial reprogramming, arterial endothelial dysfunction, and BAVM formation. We will test our hypothesis that loss of endothelial Notch gene function reprograms arteries to lose arterial and gain venous molecular identity and reduces arterial NO signaling, leading to arterial dysfunction and thus AVMs. We will analyze mice with endothelial deletion of Notch for BAVM pathology, arterial NO signaling, and endothelial response to blood flow stimuli;
Aim 3 - Ascertain the interaction between Notch and HHT in mouse dorsal aorta and cardinal vein development. We will test our hypothesis that Notch functions downstream of Hereditary Hemorrhagic Telangectasia (HHT) to mediate HHT function in AV specification. We will compare the Notch and HHT mutant phenotypes using two-photon imaging and 3D rendering and perform genetic rescue. The findings from this study will conceptually advance our understanding of the cellular and molecular mechanisms of AVM pathogenesis, reveal novel functions for Notch in regulating the unique physiology of arteries and veins, and uncover interactions between the Notch and HHT pathways. The success of this work will inspire new areas of investigation in the fields of AVMs, Notch signaling, and vascular pathophysiology.

Public Health Relevance

Brain arteriovenous malformations (BAVMs) are abnormal connections between arteries and veins that can cause stroke and epilepsy. Current treatment options are limited to surgery and radiotherapy, which are highly risky and applicable only to certain BAVM patients. This proposal is designed to determine the molecular pathways underlying BAVM formation, with the hope of identifying novel therapeutic targets to treat this disease.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
High Priority, Short Term Project Award (R56)
Project #
2R56NS067420-04
Application #
8739990
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Koenig, James I
Project Start
2009-12-01
Project End
2014-08-31
Budget Start
2013-09-30
Budget End
2014-08-31
Support Year
4
Fiscal Year
2013
Total Cost
$403,715
Indirect Cost
$146,981
Name
University of California San Francisco
Department
Surgery
Type
Schools of Medicine
DUNS #
094878337
City
San Francisco
State
CA
Country
United States
Zip Code
94143