Endoglin is required in Pax3-derived cells for embryonic blood vessel formation

Abstract

Mutations in endoglin, a TGFβ/BMP coreceptor, are causal for hereditary hemorrhagic telangiectasia (HHT). Endoglin-null (Eng−/−) mouse embryos die at embryonic day (E)10.5-11.5 due to defects in angiogenesis. In part, this is due to an absence of vascular smooth muscle cell differentiation and vessel investment. Prior studies from our lab and others have shown the importance of endoglin expression in embryonic development in both endothelial cells and neural crest stem cells. These studies support the hypothesis that endoglin may play cell-autonomous roles in endothelial and vascular smooth muscle cell precursors. However, the requirement for endoglin in vascular cell precursors remains poorly defined. Our objective was to specifically delete endoglin in neural crest- and somite-derived Pax3-positive vascular precursors to understand the impact on somite progenitor cell contribution to embryonic vascular development. Pax3Cre mice were crossed with Eng+/− mice to obtain compound mutant Pax3Cre/+;Eng+/− mice. These mice were then crossed with homozygous endoglin LoxP-mutated (EngLoxP/LoxP) mice to conditionally delete the endoglin gene in specific lineages that contribute to endothelial and smooth muscle constituents of developing embryonic vessels. Pax3Cre/+;EngLoxP/− mice showed a variety of vascular defects at E10.5, and none of these mice survived past E12.5. Embryos analyzed at E10.5 showed malformations suggestive of misdirection of the intersomitic vessels. The dorsal aorta showed significant dilation with associated vascular smooth muscle cells exhibiting disorganization and enhanced expression of smooth muscle differentiation proteins, including smooth muscle actin. These results demonstrate a requirement for endoglin in descendants of Pax3-expressing vascular cell precursors, and thus provides new insight into the cellular basis underlying adult vascular diseases such as HHT.

FACS-Seq analysis of Pax3-derived cells identifies non-myogenic lineages in the embryonic forelimb

“…Expression of certain angiogenesis-related genes can increase the rate of muscle regeneration in adult skeletal muscle 30 , 31 . Additionally, colonization of vascular cells in the developing forelimb is required for migration of Pax3 + myoblasts into the limb bud 32 , implying communication between muscle and vascular systems during development.…”

Pax3 overexpression induces cell aggregation and perturbs commissural axon projection during embryonic spinal cord development

“…Pax3 protein is involved in regulating cell aggregation and mesenchymal condensation in vitro (Wiggan et al, ). Together with Endoglin, Pax3 plays a role in embryonic angiogenesis (Young et al, ). Pax3 mutations in the human are associated with Waardenburg syndrome (type I and type III; Borycki et al, ).…”

Location, Location, Location: Signals in Muscle Specification

“…TGFβ1 also stimulates vascular SMC differentiation by activating ACTA2 [ 142 ], SM22 [ 143 ], SMAD3 [ 144 ], and RhoA [ 145 ]. TGFβ/BMP co-receptor ENDOGLIN, a descendent of the PAX3 + vascular SMCs is required for angiogenesis [ 146 ]. More recent studies also indicated the PAX3 + DM might act as a stem cell niche for vascular SMCs [ 147 ].…”