HDAC9 nuclear/cytoplasmic shuttling in pulmonary vascular endothelial barrier regulation

HDAC9核/细胞质穿梭在肺血管内皮屏障调节中的作用

基本信息

批准号：
10597538
负责人：
ALEXANDER D VERIN
金额：
$ 38.5万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-02 至 2026-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10597538
关键词：
Acceleration Acute Lung Injury Acute Respiratory Distress Syndrome Adherens Junction Affect Agonist Antibiotics Bacterial Toxins Blood Vessels Cell Nucleus Clinical Complex Cytoplasm Cytoplasmic Protein Cytoskeletal Proteins Cytosol Data Deacetylation Development Endothelial Cells Endothelium Escherichia coli Functional disorder GAB1 gene Genetic Transcription Goals Gram-Negative Bacteria Gram-Negative Bacterial Infections HDAC9 gene HGF gene Histone Deacetylase Human In Vitro Inflammation Inflammatory Response Intensive Care Investigation Knockout Mice Link Lipopolysaccharides Lung Mediating Modeling Molecular Mus Nuclear Nuclear Export Nuclear Import Pathologic Patients Permeability Phosphorylation Phosphotransferases Property Protein Dephosphorylation Protein Inhibition Protein phosphatase Proteins Pulmonary Circulation Pulmonary Edema Regulation Respiratory distress Role Sepsis Signal Transduction Source Syndrome Testing Transgenic Mice Vascular Endothelial Cell Vascular Endothelium Vascular Permeabilities analog beta catenin genetic corepressor improved in vivo lung microvascular endothelial cells mortality novel novel therapeutic intervention overexpression pharmacologic plakoglobin protective effect protein activation pulmonary function receptor

Acceleration Acute Lung Injury Acute Respiratory Distress Syndrome Adherens Junction Affect Agonist Antibiotics Bacterial Toxins Blood Vessels Cell Nucleus Clinical Complex Cytoplasm Cytoplasmic Protein Cytoskeletal Proteins Cytosol Data Deacetylation Development Endothelial Cells Endothelium Escherichia coli Functional disorder GAB1 gene Genetic Transcription Goals Gram-Negative Bacteria Gram-Negative Bacterial Infections HDAC9 gene HGF gene Histone Deacetylase Human In Vitro Inflammation Inflammatory Response Intensive Care Investigation Knockout Mice Link Lipopolysaccharides Lung Mediating Modeling Molecular Mus Nuclear Nuclear Export Nuclear Import Pathologic Patients Permeability Phosphorylation Phosphotransferases Property Protein Dephosphorylation Protein Inhibition Protein phosphatase Proteins Pulmonary Circulation Pulmonary Edema Regulation Respiratory distress Role Sepsis Signal Transduction Source Syndrome Testing Transgenic Mice Vascular Endothelial Cell Vascular Endothelium Vascular Permeabilities analog beta catenin genetic corepressor improved in vivo lung microvascular endothelial cells mortality novel novel therapeutic intervention overexpression pharmacologic plakoglobin protective effect protein activation pulmonary function receptor

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项目摘要

PROJECT SUMMARY Acute lung injury (ALI) is characterized by lung vascular endothelial (EC) barrier compromise resulting in pulmonary edema. Gram negative (G-) bacteria are the source of bacterial toxins such as lipopolysaccharides (LPS), which are potent triggers of ALI. Despite the use of potent antibiotics and aggressive intensive care support, the mortality of LPS-induced ALI remains high, primarily because the complex molecular mechanisms involved in EC barrier regulation are ill-defined. The Scientific Premise of this Project derives from our novel preliminary data indicating that the class IIa histone deacetylase, HDAC9, is upregulated in the lungs of patients with sepsis in concert with the inflammatory responses. Accordingly, the overexpression of HDAC9 in transgenic mice increases basal lung vascular permeability and exacerbates vascular barrier compromise induced by LPS. Conversely, deletion of HDAC9 gene improves lung function in LPS-induced murine ALI model. These data suggest pro-edemagenic role of HDAC9 expression in ALI. HDAC9 is nuclear-cytoplasmic protein and its cellular localization is regulated by phosphorylation. Phosphorylation leads to HDAC9 nuclear export thus inhibiting its activity in the nucleus and promoting interactions with extra-nuclear targets. We demonstrated that LPS increases HDAC9 phosphorylation accompanied by its nuclear export in EC. In contrary, de-phosphorylation of HDAC9 leads to nuclear import thus inhibiting its association with cytoplasmic proteins. We showed that HDAC9 interacts with protein phosphatase 2A (PP2A), which has a barrier-protective role in EC. Further, our novel data indicated that PP2A is activated by Gi-mediated EC barrier-protective agonists, HGF and the stable ATP analog, ATPγS, suggesting the link between Gi-mediated signaling and PP2A in EC barrier enhancement. While the mechanisms of Gi-mediated PP2A activation have not been described, our data suggest that they may include the activation of GAB1/Shp2-mediated signaling. Further, both HDAC9 and PP2A interact with AJ protein plakoglobin suggesting functional complex. However, whether the HDAC9-dependent EC barrier regulation involved HDAC9/PP2A/plakoglobin interaction and EC barrier-protective effect of PP2A is mediated through decreased HDAC9 phosphorylation is not yet established. Therefore, a primary goal of this Proposal will be to define a novel role for HDAC9/PP2A crosstalk in AJ-mediated EC barrier regulation. We hypothesize that the phosphorylation status of HDAC9 and its interaction with PP2A alter intracellular signaling to regulate the EC barrier via effect on AJ assembly. The SAs are: 1: To evaluate the role of HDAC9 phosphorylation/nuclear export in the LPS-induced adherens junctions-mediated endothelial barrier compromise in vitro and in vivo. 2: To determine the importance of PP2A-mediated HDAC9 dephosphorylation/nuclear import in the assembly of AJs and endothelial barrier strengthening in vitro and in vivo.

项目摘要急性肺损伤（ALI）的特征是肺血管内皮（EC）屏障妥协，导致肺水肿。革兰氏阴性（G-）细菌是细菌毒素（如脂多糖）的来源（LPS），是Ali的潜在触发器。尽管使用了潜在的抗生素和侵略性重症监护支持，LPS诱导的ALI的死亡率仍然很高，主要是因为复杂的分子机制涉及EC屏障调节的定义不明。该项目的科学前提源自我们的小说初步数据表明IIA类组蛋白脱乙酰基酶HDAC9在患者的肺中进行了更新败血症与炎症反应一致。根据转基因中HDAC9的过表达小鼠增加了基本的肺血管通透性，并加剧了LPS损害的血管屏障。相反，HDAC9基因的缺失改善了LPS诱导的鼠ALI模型中的肺功能。这些数据建议HDAC9在ALI中表达的促氧化昆虫作用。 HDAC9是核质蛋白及其细胞定位受磷酸化调节。磷酸化导致HDAC9核输出抑制其细胞核的活性并促进与核外靶标相互作用。我们证明了LPS 增加HDAC9磷酸化，并伴随其在EC中的核出口。相反，去磷酸化 HDAC9导致核进口，从而抑制其与细胞质蛋白的关联。我们证明了HDAC9 与蛋白质磷酸酶2a（PP2A）相互作用，该蛋白在EC中具有屏障保护作用。此外，我们的新数据表明PP2A被GI介导的EC屏障保护激动剂HGF和稳定的ATP类似物激活 ATPγs，表明在EC屏障增强中，GI介导的信号传导与PP2A之间的联系。而 GI介导的PP2A激活的机制尚未描述，我们的数据表明它们可能包括 GAB1/SHP2介导的信号传导的激活。此外，HDAC9和PP2A都与AJ蛋白相互作用 Plakoglobin提示功能复合物。但是，是否依赖HDAC9依赖性EC屏障调节涉及hDAC9/pp2a/plakoglobin的相互作用和PP2A的EC屏障保护作用是通过尚未确定HDAC9磷酸化降低。因此，该提议的主要目标是定义了AJ介导的EC屏障调节中HDAC9/PP2A串扰的新作用。我们假设 HDAC9的磷酸化状态及其与PP2A的相互作用改变细胞内信号传导以调节EC 通过对AJ组件的效应障碍。 SAS是：1：评估HDAC9磷酸化/核输出的作用在LPS诱导的粘附剂连接介导的内皮屏障在体外和体内造成的损害。 2：到确定PP2A介导的HDAC9去磷酸化/核进口在AJS组装中的重要性和内皮屏障在体外和体内增强。