CD45 足以启动人内皮细胞的内皮细胞向间充质转化-简短报告。
CD45 Is Sufficient to Initiate Endothelial-to-Mesenchymal Transition in Human Endothelial Cells-Brief Report.
机构信息
Vascular Biology Program and Department of Surgery (S.N., J.W.-S., Q.P., B.Z., H.C., J.B.), Boston Children's Hospital, MA.
Departments of Surgery (S.N., Q.P., B.Z., H.C., J.B.), Harvard Medical School, Boston, MA.
出版信息
Arterioscler Thromb Vasc Biol. 2023 May;43(5):e124-e131. doi: 10.1161/ATVBAHA.122.318172. Epub 2023 Mar 16.
BACKGROUND
Endothelial-to-mesenchymal transition (EndMT) is a dynamic process in which endothelial cells acquire mesenchymal properties and in turn contribute to tissue remodeling and growth. Previously, we found EndMT associated with mitral valve adaptation after myocardial infarction. Furthermore, mitral valve endothelial cells collected at 6 months post-myocardial infarction expressed the pan-leukocyte marker CD45 and EndMT markers. Additionally, mitral valve endothelial cells induced to undergo EndMT with TGF (transforming growth factor)-β1 strongly coexpressed CD45 but not CD11b or CD14. Pharmacologic inhibition of the CD45 PTPase (protein tyrosine phosphatase) domain in mitral valve endothelial cells blocked TGFβ-induced EndMT. This prompted us to speculate that, downstream of TGFβ, CD45 induces EndMT.
METHODS
We activated the endogenous CD45 promoter in human endothelial colony forming cells (ECFCs) using CRISPR (cluster regularly interspaced short palindromic repeats)/inactive Cas9 (CRISPR-associated protein 9) transcriptional activation. Bulk RNA sequencing was performed on control ECFCs and CD45-positive ECFCs to identify transcriptomic changes. Three functional assays-cellular migration, collagen gel contraction, and transendothelial electrical resistance-were conducted to assess mesenchymal properties in CD45-positive ECFCs.
RESULTS
Activation of the endogenous CD45 promoter in ECFC and 3 additional sources of endothelial cells induced expression of several genes implicated in EndMT. In addition, CD45-positive ECFCs showed increased migration, a hallmark of EndMT, increased collagen gel contraction, a hallmark of mesenchymal cells, and decreased cell-cell barrier integrity, indicating reduced endothelial function.
CONCLUSIONS
CD45 is sufficient to incite an EndMT phenotype and acquisition of mesenchymal cell properties in normal human ECFCs. We speculate that CD45, through its C-terminal PTPase domain, initiates signaling events that drive EndMT.
背景
内皮细胞向间充质转化（EndMT）是一个动态过程，在此过程中内皮细胞获得间充质特性，并转而促进组织重塑和生长。之前，我们发现 EndMT 与心肌梗死后二尖瓣的适应性改变有关。此外，在心肌梗死后 6 个月采集的二尖瓣内皮细胞表达泛白细胞标志物 CD45 和 EndMT 标志物。此外，用 TGF（转化生长因子）-β1 诱导二尖瓣内皮细胞发生 EndMT 时，强烈共表达 CD45，但不表达 CD11b 或 CD14。在二尖瓣内皮细胞中抑制 CD45 PTP 酶（蛋白酪氨酸磷酸酶）结构域可阻断 TGFβ 诱导的 EndMT。这促使我们推测，在 TGFβ 下游，CD45 诱导 EndMT。
方法
我们使用 CRISPR（簇状规律间隔短回文重复序列）/无活性 Cas9（CRISPR 相关蛋白 9）转录激活来激活人内皮集落形成细胞（ECFC）中的内源性 CD45 启动子。对对照 ECFC 和 CD45 阳性 ECFC 进行批量 RNA 测序，以鉴定转录组变化。进行了 3 种功能测定——细胞迁移、胶原凝胶收缩和跨内皮电阻——以评估 CD45 阳性 ECFC 中的间充质特性。
结果
在 ECFC 和另外 3 种内皮细胞来源中激活内源性 CD45 启动子可诱导几种参与 EndMT 的基因表达。此外，CD45 阳性 ECFC 显示出增加的迁移，这是 EndMT 的一个标志，增加的胶原凝胶收缩，这是间充质细胞的一个标志，以及降低的细胞-细胞屏障完整性，表明内皮功能降低。
结论
CD45 足以在正常人类 ECFC 中引发 EndMT 表型和获得间充质细胞特性。我们推测，CD45 通过其 C 端 PTP 酶结构域启动驱动 EndMT 的信号事件。
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