Essential Role of Endothelial Smad4 in Vascular Remodeling and Integrity

Abstract

New blood vessels are formed through the assembly or sprouting of endothelial cells (ECs) and become stabilized by the formation of perivascular matrix and the association with supporting mural cells. To investigate the role of endothelial Smad4 in vascular development, we deleted the Smad4 gene specifically in ECs using the Cre-LoxP system. EC-specific Smad4 mutant mice died at embryonic day 10.5 due to cardiovascular defects, including attenuated vessels sprouting and remodeling, collapsed dorsal aortas, enlarged hearts with reduced trabeculae, and failed endocardial cushion formation. Noticeably, Smad4-deficient ECs demonstrated an intrinsic defect in tube formation in vitro. Furthermore, the mutant vascular ECs dissociated away from the surrounding cells and suffered from impaired development of vascular smooth muscle cells. The disturbed vascular integrity and maturation was associated with aberrant expression of angiopoietins and a gap junction component, connexin43. Collectively, we have provided direct functional evidence that Smad4 activity in the developing ECs is essential for blood vessel remodeling, maturation, and integrity.

This study was supported by the National Key Basic Research Program of China (2005CB522506, 2005CB522705, and 2006CB943501), the National Natural Science Foundation of China (30671077), the National Hi-Tech R&D Program (2006AA02Z168), the National Science and Technology Supporting Program (2006BAI23B01-03), and Beijing Science Projects (10006303041231).

We thank Chuxia Deng at the National Institutes of Health (NIH) for the Smad4 conditional gene targeting mouse, Hua Gu at the NIH for Cre plasmid, and Xiang Gao at Nanjing University for the mouse strains. We also thank Jincai Luo at Peking University for instruction on selective transformation of ECs and related retrovirus and cell lines, and we thank Yeguang Chen at Tsinghua University for reporters.