Vascular Dysfunction and Inflammation

血管功能障碍和炎症

• 批准号: 10923692
• 负责人: ROBERT L DANNER
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10923692
• 关键词: 2019-nCoV; 3-Dimensional; ACE2; Acute; Acute Respiratory Distress Syndrome; Affect; Agonist; Aldosterone; Apoptosis; Arginine; Atherosclerosis; Award; BMPR2 gene; Biological Models; Biomedical Engineering; Blood; Blood Vessels; COVID-19; COVID-19 complications; COVID-19 susceptibility; Cardiogenic Shock; Cardiovascular system; Cells; Cessation of life; Chronic; Circulation; Complex; Complication; Coronavirus; Cyclic AMP-Dependent Protein Kinases; Cyclic GMP; Cytoskeleton; DNA Sequence; Disease; Disease Progression; Drug Targeting; EP300 gene; ERCC3 gene; Endothelium; Event; Expression Profiling; Family member; Female; Festival; Fibroblasts; Fish Oils; Flow Cytometry; G-Protein-Coupled Receptors; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Predisposition to Disease; Genetic Transcription; Genomics; Goals; Growth; HIV; Heart; Hemostatic function; Human; IL8 gene; Immunochemistry; Impairment; In Vitro; Individual; Inflammation; Inflammatory; Inflammatory Response; Infusion procedures; Injury; Interferons; Interruption; Investigation; Iron; Laboratory Study; Left; Link; Lung; MAP Kinase Gene; MAPK3 gene; MAPK8 gene; Malignant Neoplasms; Mediating; Mediator; Mesenchymal; Messenger RNA; Meta-Analysis; Metabolism; Mitochondria; Modeling; Molecular Conformation; Monounsaturated Fatty Acids; Multiple Organ Failure; Mus; Mutation; Myocardial Infarction; NF-kappa B; NOS3 gene; NOTCH1 gene; Natural History; Nitric Oxide; Nitric Oxide Signaling Pathway; Nitric Oxide Synthase; Nitric Oxide Synthase Type I; Nuclear Receptors; Nucleic Acids; Outcome; Oxidants; Oxygen; PI3K/AKT; PPAR gamma; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Peroxisome Proliferator-Activated Receptors; Pharmaceutical Preparations; Phenotype; Phosphorylation; Pilot Projects; Play; Polyunsaturated Fatty Acids; Preparation; Procollagen-Proline Dioxygenase; Production; Proliferating; Proteins; Proto-Oncogene Proteins c-akt; Pulmonary Hypertension; Ras/Raf; Rattus; Reactive Oxygen Species; Receptor Signaling; Refractory; Relaxation; Repression; Research; Research Personnel; Resistance; Risk Factors; Role; STAT1 gene; Safety; Scleroderma; Sepsis; Septic Shock; Sickle Cell Anemia; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Spironolactone; Stress; Structure of parenchyma of lung; System; TMPRSS2 gene; Tertiary Protein Structure; Therapeutic; Therapeutic Intervention; Transcript; Transcription Factor AP-1; Translations; United States National Institutes of Health; Variant; Vascular Diseases; Vascular remodeling; Ventricular End-Diastolic Volumes; Viral Proteins; Virus; Vision; Work; alpha Tubulin; antagonist; apoAI regulatory protein-1; arteriole; bench to bedside; cardiometabolic risk; cardiovascular risk factor; caveolin 1; cohort; constriction; dietary; endothelial dysfunction; heart preservation; hemodynamics; indexing; inhibitor; knock-down; loss of function; loss of function mutation; lung injury; lung vascular injury; mortality; mouse model; organ injury; overexpression; p38 Mitogen Activated Protein Kinase; phase 3 testing; prevent; primary pulmonary hypertension; prototype; pulmonary arterial hypertension; repaired; right ventricular failure; rosiglitazone; senescence; severe COVID-19; small molecule inhibitor; stroke risk; symposium; therapeutic target; thrombotic; transcription factor TFIIH; vascular bed; vascular inflammation

Nitric oxide (NO): NO upregulates TNFa production (J Immunol 1994; Blood 1997) through a cGMP-independent pathway (J Biol Chem 1997; J Biol Chem 1999; J Biol Chem 2003), while ROS from eNOS uncoupling upregulates TNFa (J Biol Chem 2000) through ERK1/2 (Am J Physiol 2001). NO activation of p38 MAPK stabilizes IL-8 mRNA (J Infect Dis 1998; J Leuk Biol 2004). NO has diverse effects on transcript stability and translation (Nucleic Acids Research 2006; J Leuk Biol 2008). Sickle cell disease causes oxidant and inflammatory stress in the vasculature (Blood, 2004), altering gene expression and arginine metabolism (Circulation, 2007). NO activation of p38 MAPK stabilized p21 mRNA and was antiproliferative (BMC Genomics 2005; J Biol Chem 2006). NO activated PPARg through p38 MAPK (FASEB J 2007) protecting the endothelium. Unlike NO, CO blocked NF-kB signaling, broadly suppressing inflammation (PLoS One 2009). Nuclear receptors (NRs): G-protein coupled receptor 40 (GPR40)/p38 MAPK/PGC1a/EP300 activation by rosiglitazone (RGZ) was shown to augment RGZ/PPARg genomic signaling (J Biol Chem 2015). Cognate GPR and nuclear receptor signaling networks may explain differences in the safety and efficacy of NR targeted drugs (Pharm Research 2016). Long-chain monounsaturated fatty acids (LCMUFA; i.e., C20:1 and C22:1) benefits were associated with PPAR activation, via GPR40 activation (Atherosclerosis 2017). MR agonists repressed NF-kB mediated gene transcription, but trans-activated AP-1 signaling in a DNA sequence, MR conformation, and AP-1 family member dependent fashion (J Biol Chem 2016). Aldosterone/MR activation of AP-1 contribute to harm in CHF and PAH. Spironolactone (SPL) suppresses both NF-kB and AP-1 inflammatory signaling independent of MR through proteasomal degradation of XPB, a subunit of the TFIIH transcription complex (Cardiovasc Res 2018). CFH infusions resulted in pulmonary hypertension, cardiogenic shock, and multiorgan failure, likely through NO scavenging. During sepsis, CFH infusions worsened oxygen exchange and lung injury, presumably by supplying iron that promoted bacterial growth. CFH elevation in septic shock adversely impacts sepsis outcomes through more than one mechanism that could be therapeutically targeted (Am J Physiol Heart Circ Physiol 2021). Pulmonary arterial hypertension (PAH): A pilot study of SPL therapy (Trials 2013) and a natural history study investigating vascular inflammation support ongoing laboratory studies. Circulating ECs were identified and validated using flow cytometry and ultramicro analytical immunochemistry (Thromb Haemostasis 2014). Loss-of-function mutations in bone morphogenetic protein type II receptor (BMPR2) are the most common genetic cause of PAH. BMPR2 knockdown (KD) in human PAECs activated Ras/Raf/ERK signaling leading to proliferation, invasiveness, and cytoskeletal abnormalities (Am J Physiol Lung Cell Mol Physiol 2016). A meta-analysis of PBMC expression profiling in PAH patients identified IFN-driven inflammation as a fundamental component of PAH pathobiology that was unrecognized in individual blood profiling studies (Am J Physiol Lung Cell Mol Physiol 2020). Caveolin-1 (CAV1) loss-of-function (LOF), similar to BMPR2, produced a proliferative, hyper-migratory and inflammatory PAEC phenotype associated with JAK/STAT/interferon and AKT activation. This inflammatory signature was also found in fibroblasts from PAH patients with CAV1 mutations and in CAV1-/- mice. CAV1 loss and STAT1 activation was also seen in the pulmonary arterioles of patients with idiopathic PAH. Blocking JAK/STAT or AKT rescued aspects of CAV1 loss, only AKT inhibitors suppressed activation of both signaling pathways. Silencing endothelial nitric oxide synthase (NOS3) prevented STAT1 and AKT activation induced by CAV1 loss, implicating CAV1/NOS3 uncoupling in the inflammatory phenotype associated with CAV1 loss (Proc Natl Acad Sci USA 2021). MR antagonist treatment in the SuHx rat model of PAH preserved cardiac index and increased left ventricular (LV) end-diastolic volume index. EPL treatment blunted the induction of MR target and inflammatory response genes in the RV (Am J Physiol Lung Cell Mol Physiol 2022). In our CAV1 loss model of PAH, NOS3 co-silencing not only blocked STAT1 and NOS3 phosphorylation, but also diminished ROS generation. Small molecule inhibitors of sAC and PKA blocked NOS3 and STAT1 phosphorylation suggesting a possible role for this pathway in CAV1 loss associated abnormalities (ATS abstract 2022). Neither pseudotyped nor live SARS-CoV-2 virus appear to readily infect or replicate in human pulmonary ECs in vitro, with the exception of the D614G variant. Low expression of ACE2 and TMPRSS2 in PAECs may explain their diminished susceptibility to SARS-CoV-2 (ATS abstract 2022). LOF mutations in COUPTF2 (NR2F2) have been associated with CHD, which can result in PAH. COUPTF2 silencing in ECs produced an IFN inflammatory response and a hyper-proliferative, apoptosis-resistant, and invasive phenotype. Dickkopf-1 (DKK1) was induced by COUPTF2 loss and DKK1 knockdown abrogated signaling and phenotypic abnormalities (Am J Physiol Lung Cell Mol Physiol 2023). An in vitro pseudohypoxia model of PAH was established by silencing PHD2 (prolyl hydroxylase domain protein 2; EGLN1) in LMVECs. PHD2-silencing stabilized HIF2alpha, decreased ASK-interacting protein 1 (AIP; DAB2IP), and activated AKT and ERK (Aspen Lung Conference 2019; MS submission pending 2023). Marked resistance to apoptosis has been a consistent feature of our EC models of PAH. Using the BMPR2 LOF model as a prototype, apoptosis resistance was linked to DLL4/NOTCH1 signaling loss with PI3K/AKT activation, and JNK suppression (Aspen Lung Conference 2019). Importantly, DLL4 loss was seen across several of our models of PAH and validated in lung tissue from iPAH patients. Blocking PI3K/AKT or PPARgamma overexpression restored apoptosis sensitivity in three model systems, BMPR2, CAV1 and PHD2 (ATS abstract 2023). Interactions among BMPR2, DLL4/NOTCH1/PPAR, and PI3K/AKT may lead to targeted approaches for treating vascular remodeling in PAH (MS pending submission 09/2023). Vasohibin-1 (VASH1) loss with increased alpha-tubulin tyrosination was implicated in BMPR2 loss-associated cytoskeletal abnormalities and endothelial dysfunction (MS in preparation 2024). SMAD9 LOF in human PAECs also produced an abnormal cellular phenotype characterized by proliferation, hypermigration, cytoskeletal and mitochondrial alterations and endothelial to mesenchymal transition, as well as non-canonical activation of AKT, ERK and p38 (ATS 2018; MS in preparation). COVID-19 has acute and chronic manifestations. Endothelial senescence may underlie the thrombotic microvasculopathy associated with severe COVID-19 as well as the increased risk of cardiovascular events in patients who have otherwise recovered. We have launched a multi-institute project to investigate this hypothesis using deeply phenotyped patient cohorts, a bioengineered three-dimensional disease-on-chip in vitro system and a coronavirus mouse model. Preliminary results have demonstrated that SARS-CoV-2 viral proteins are readily taken up by human endothelium and trigger a senescent cellular phenotype (NIH Research Festival 2023; Fellows Award for Research Excellence (FARE) 2024). Endothelial senescence in this model system is reversable using a drug undergoing phase III testing in acute COVID-19. Associate Investigator on a 2023 Bench to Bedside application: Effect of dietary fish oil enriched in very-long-chain polyunsaturated fatty acids (VLCPUFA) on cardiometabolic risk factors and visual function. Principle Investigator on a 2023 Bench to Bedside application: Endothelial senescence in acute and late covid-19 vasculopathy.

一氧化氮 (NO)：NO 通过 cGMP 独立途径（J Biol Chem 1997；J Biol Chem 1999；J Biol Chem 2003）上调 TNFa 的产生（J Nutrition 1994；Blood 1997），而 eNOS 解偶联产生的 ROS 上调 TNFa（J Biol Chem 1997；J Biol Chem 1999；J Biol Chem 2003）。 Biol Chem 2000) 至 ERK1/2 (Am J生理学 2001）。 p38 MAPK 的 NO 激活可稳定 IL-8 mRNA（J Infect Dis 1998；J Leuk Biol 2004）。 NO 对转录稳定性和翻译有多种影响（Nucleic Acids Research 2006；J Leuk Biol 2008）。 镰状细胞病会导致脉管系统中的氧化和炎症应激（Blood，2004），改变基因表达和精氨酸代谢（Circulation，2007）。 p38 MAPK 的 NO 激活可稳定 p21 mRNA，并具有抗增殖作用（BMC Genomics 2005；J Biol Chem 2006）。 NO 通过 p38 MAPK (FASEB J 2007) 激活 PPARg 保护内皮。与 NO 不同，CO 阻断 NF-kB 信号传导，广泛抑制炎症 (PLoS One 2009)。 核受体 (NR)：罗格列酮 (RGZ) 激活 G 蛋白偶联受体 40 (GPR40)/p38 MAPK/PGC1a/EP300 可增强 RGZ/PPARg 基因组信号传导 (J Biol Chem 2015)。同源 GPR 和核受体信号网络可能解释 NR 靶向药物的安全性和有效性差异（Pharm Research 2016）。长链单不饱和脂肪酸（LCMUFA；即 C20:1 和 C22:1）的益处与 PPAR 通过 GPR40 激活相关（Atherosclerosis 2017）。 MR 激动剂抑制 NF-kB 介导的基因转录，但以 DNA 序列、MR 构象和 AP-1 家族成员依赖性方式反式激活 AP-1 信号传导 (J Biol Chem 2016)。 AP-1 的醛固酮/MR 激活会导致 CHF 和 PAH 的危害。螺内酯 (SPL) 通过 XPB（TFIIH 转录复合物的一个亚基）的蛋白酶体降解，抑制独立于 MR 的 NF-kB 和 AP-1 炎症信号传导 (Cardiovasc Res 2018)。 CFH 输注可能通过 NO 清除导致肺动脉高压、心源性休克和多器官衰竭。在脓毒症期间，CFH 输注会加剧氧交换和肺损伤，可能是通过提供促进细菌生长的铁来实现的。脓毒性休克中的 CFH 升高通过不止一种可作为治疗目标的机制对脓毒症结局产生不利影响 (Am J Physiol Heart Circ Physiol 2021)。 肺动脉高压 (PAH)：SPL 疗法的初步研究（2013 年试验）和调查血管炎症的自然史研究支持正在进行的实验室研究。使用流式细胞术和超微分析免疫化学对循环 EC 进行鉴定和验证（Thromb Haemostasis 2014）。 II 型骨形态发生蛋白受体 (BMPR2) 的功能丧失突变是 PAH 最常见的遗传原因。人 PAEC 中的 BMPR2 敲低 (KD) 激活 Ras/Raf/ERK 信号传导，导致增殖、侵袭和细胞骨架异常 (Am J Physiol Lung Cell Mol Physiol 2016)。 对 PAH 患者 PBMC 表达谱的荟萃分析发现，IFN 驱动的炎症是 PAH 病理学的基本组成部分，而个体血液谱研究并未认识到这一点 (Am J Physiol Lung Cell Mol Physiol 2020)。 Caveolin-1 (CAV1) 功能丧失 (LOF) 与 BMPR2 类似，产生与 JAK/STAT/干扰素和 AKT 激活相关的增殖、过度迁移和炎症 PAEC 表型。在携带 CAV1 突变的 PAH 患者和 CAV1-/- 小鼠的成纤维细胞中也发现了这种炎症特征。在特发性 PAH 患者的肺小动脉中也发现了 CAV1 缺失和 STAT1 激活。阻断 JAK/STAT 或 AKT 可以挽救 CAV1 损失的各个方面，只有 AKT 抑制剂才能抑制两条信号通路的激活。沉默内皮一氧化氮合酶 (NOS3) 可阻止 CAV1 缺失引起的 STAT1 和 AKT 激活，这表明与 CAV1 缺失相关的炎症表型中 CAV1/NOS3 解偶联 (Proc Natl Acad Sci USA 2021)。 SuHx 大鼠 PAH 模型中的 MR 拮抗剂治疗保留了心脏指数并增加了左心室 (LV) 舒张末期容积指数。 EPL 治疗削弱了 RV 中 MR 靶点和炎症反应基因的诱导 (Am J Physiol Lung Cell Mol Physiol 2022)。 在我们的 PAH 的 CAV1 缺失模型中，NOS3 共沉默不仅阻断了 STAT1 和 NOS3 磷酸化，而且还减少了 ROS 的产生。 sAC 和 PKA 的小分子抑制剂可阻断 NOS3 和 STAT1 磷酸化，表明该通路在 CAV1 丢失相关异常中可能发挥作用（ATS 摘要 2022）。 除 D614G 变种外，假型和活 SARS-CoV-2 病毒似乎都不容易在体外感染人肺 EC 或在 EC 中复制。 PAEC 中 ACE2 和 TMPRSS2 的低表达可能解释了它们对 SARS-CoV-2 敏感性降低的原因（ATS 摘要 2022）。 COUPTF2 (NR2F2) 中的 LOF 突变与 CHD 相关，而 CHD 可能导致 PAH。 ECs 中的 COUPTF2 沉默会产生 IFN 炎症反应以及过度增殖、抗凋亡和侵袭表型。 Dickkopf-1 (DKK1) 是由 COUPTF2 缺失和 DKK1 敲低消除信号传导和表型异常诱导的 (Am J Physiol Lung Cell Mol Physiol 2023)。 通过沉默 LMVEC 中的 PHD2（脯氨酰羟化酶结构域蛋白 2；EGLN1）建立 PAH 体外假性缺氧模型。 PHD2 沉默可稳定 HIF2α，减少 ASK 相互作用蛋白 1（AIP；DAB2IP），并激活 AKT 和 ERK（2019 年阿斯彭肺会议；MS 提交待 2023 年）。 显着的细胞凋亡抗性是我们的 PAH EC 模型的一致特征。使用 BMPR2 LOF 模型作为原型，细胞凋亡抵抗与 DLL4/NOTCH1 信号丢失、PI3K/AKT 激活和 JNK 抑制相关（Aspen Lung Conference 2019）。重要的是，我们的几个 PAH 模型中都发现了 DLL4 丢失，并在 iPAH 患者的肺组织中得到了验证。阻断 PI3K/AKT 或 PPARgamma 过表达可恢复 BMPR2、CAV1 和 PHD2 三个模型系统中的细胞凋亡敏感性（ATS 摘要 2023）。 BMPR2、DLL4/NOTCH1/PPAR 和 PI3K/AKT 之间的相互作用可能会导致治疗 PAH 血管重塑的靶向方法（MS 待提交 09/2023）。 Vasohibin-1 (VASH1) 损失伴随 α-微管蛋白酪氨酸化增加，与 BMPR2 损失相关的细胞骨架异常和内皮功能障碍有关（MS 准备中 2024）。人 PAEC 中的 SMAD9 LOF 还产生异常细胞表型，其特征是增殖、过度迁移、细胞骨架和线粒体改变、内皮向间质转化，以及 AKT、ERK 和 p38 的非典型激活（ATS 2018；MS 准备中）。 COVID-19 有急性和慢性表现。内皮衰老可能是与严重 COVID-19 相关的血栓性微血管病变的基础，以及已康复患者心血管事件风险增加的原因。我们启动了一个多机构项目，利用深度表型患者队列、生物工程三维芯片疾病体外系统和冠状病毒小鼠模型来研究这一假设。初步结果表明，SARS-CoV-2 病毒蛋白很容易被人类内皮细胞吸收并引发衰老细胞表型（NIH 研究节 2023；研究员卓越研究奖 (FARE) 2024）。使用在急性 COVID-19 中进行 III 期测试的药物，该模型系统中的内皮衰老是可逆转的。 2023 年实验室应用的副研究员：富含极长链多不饱和脂肪酸 (VLCPUFA) 的膳食鱼油对心脏代谢危险因素和视觉功能的影响。 2023 年实验室应用的原理研究员：急性和晚期 covid-19 血管病变中的内皮衰老。

项目成果

Beneficial effects of stress-dose corticosteroid therapy in canines depend on the severity of staphylococcal pneumonia.

应激剂量皮质类固醇治疗对犬科动物的有益效果取决于葡萄球菌肺炎的严重程度。

• 发表时间: 2012-12
• 影响因子: 38.9
• 作者: Hicks, Caitlin W;Sweeney, Daniel A;Danner, Robert L;Eichacker, Peter Q;Suffredini, Anthony F;Feng, Jing;Sun, Junfeng;Moriyama, Brad;Wesley, Robert;Behrend, Ellen N;Solomon, Steven B;Natanson, Charles
• 通讯作者: Natanson, Charles

Nitric oxide-p38 MAPK signaling stabilizes mRNA through AU-rich element-dependent and -independent mechanisms.

一氧化氮-p38 MAPK 信号通过富含 AU 元素的依赖和独立机制稳定 mRNA。

• 发表时间: 2008-04
• 影响因子: 5.5
• 作者: Wang, Shuibang;Zhang, Jianhua;Zhang, Yi;Kern, Steven;Danner, Robert L
• 通讯作者: Danner, Robert L

Meta-analysis of blood genome-wide expression profiling studies in pulmonary arterial hypertension.

肺动脉高压血液全基因组表达谱研究的荟萃分析。

• 发表时间: 2020-01-01
• 作者: Elinoff, Jason M;Mazer, Adrien J;Cai, Rongman;Lu, Mengyun;Graninger, Grace;Harper, Bonnie;Ferreyra, Gabriela A;Sun, Junfeng;Solomon, Michael A;Danner, Robert L
• 通讯作者: Danner, Robert L

In critically ill patients with COVID-19, antiplatelet therapy did not increase organ support-free days at 21 d.

在患有 COVID-19 的危重患者中，抗血小板治疗并没有增加 21 天时的器官无支持天数。

• 发表时间: 2022
• 影响因子: 39.2
• 作者: Athale, Janhavi;Danner, Robert L
• 通讯作者: Danner, Robert L

Isolation of a circulating CD45-, CD34dim cell population and validation of their endothelial phenotype.

分离循环 CD45-、CD34dim 细胞群并验证其内皮表型。

• 发表时间: 2014-10
• 影响因子: 6.7
• 作者: Tropea, Margaret M;Harper, Bonnie J A;Graninger, Grace M;Phillips, Terry M;Ferreyra, Gabriela;Mostowski, Howard S;Danner, Robert L;Suffredini, Anthony F;Solomon, Michael A
• 通讯作者: Solomon, Michael A