NOTCH3 expression is induced in mural cells through an autoregulatory loop that requires endothelial-expressed JAGGED1.
NOTCH3 expression is induced in mural cells through an autoregulatory loop that requires endothelial-expressed JAGGED1.
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DOI：
10.1161/circresaha.108.184846
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发表时间：
2009-02-27
影响因子：
20.1
通讯作者：
Lilly B
中科院分区：
文献类型：
--
作者：
Liu H;Kennard S;Lilly B
已知内皮细胞和壁细胞（平滑肌细胞、周细胞或成纤维细胞）彼此通信。它们的相互作用不仅用于支持功能齐全的血管，而且还可以调节血管组装和分化或成熟。为了更好地理解这种异型相互作用的分子组成部分，我们利用了血管生成的三维模型，并筛选了通过共培养这两种不同细胞类型来调节的基因。在这样做的过程中，我们发现NOTCH 3是一种通过与内皮细胞共培养在壁细胞中强烈诱导表达的基因。通过siRNA敲低表明，NOTCH 3是内皮依赖性壁细胞分化所必需的，而过表达NOTCH 3足以促进平滑肌基因表达。此外，NOTCH 3有助于与内皮细胞共培养的壁细胞的促血管生成能力。有趣的是，我们发现NOTCH 3的表达依赖于Notch信号传导，因为γ-分泌酶抑制剂DAPT阻断了其上调。此外，在壁细胞中，显性负性Mastermind样1构建体抑制NOTCH 3表达，并且其诱导需要内皮表达的JAGGED 1。此外，我们证明了NOTCH 3可以促进其自身的表达和JAGGED 1在壁细胞中的表达。总之，这些数据提供了一种机制，通过该机制，内皮细胞通过激活和诱导NOTCH 3诱导壁细胞分化。这些发现还表明，NOTCH 3具有通过包括自身调节和JAGGED 1表达的正反馈回路维持分化表型的能力。
Endothelial cells and mural cells (smooth muscle cells, pericytes, or fibroblasts) are known to communicate with one another. Their interactions not only serve to support fully functional blood vessels, but also can regulate vessel assembly and differentiation or maturation. In an effort to better understand the molecular components of this heterotypic interaction, we utilized a 3-dimensional model of angiogenesis and screened for genes, which were modulated by coculturing of these two different cell types. In doing so, we discovered that NOTCH3 is one gene whose expression is robustly induced in mural cells by coculturing with endothelial cells. Knockdown by siRNA revealed that NOTCH3 is necessary for endothelial-dependent mural cell differentiation, whereas overexpression of NOTCH3 is sufficient to promote smooth muscle gene expression. Moreover, NOTCH3 contributes to the proangiogenic abilities of mural cells cocultured with endothelial cells. Interestingly, we found that the expression of NOTCH3 is dependent upon Notch signaling, as the γ-secretase inhibitor DAPT blocked its upregulation. Furthermore, in mural cells, a dominant-negative Mastermind-like1 construct inhibited NOTCH3 expression, and endothelial-expressed JAGGED1 was required for its induction. Additionally, we demonstrated that NOTCH3 could promote its own expression and that of JAGGED1 in mural cells. Taken together, these data provide a mechanism by which endothelial cells induce the differentiation of mural cells through activation and induction of NOTCH3. These findings also suggest that NOTCH3 has the capacity to maintain a differentiated phenotype through a positive feedback loop that includes both autoregulation and JAGGED1 expression.