Targeting Pathogenic Endothelial Dysfunction in Lupus Nephritis

针对狼疮性肾炎的致病性内皮功能障碍

• 批准号: 10487863
• 负责人: JAMES C OATES
• 金额: --
• 依托单位: RALPH H JOHNSON VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487863
• 关键词: Address; Adhesions; Affect; Anti-Inflammatory Agents; Antigen-Antibody Complex; Arteries; Atherosclerosis; Biological Markers; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cells; Chemotaxis; Cholesterol; Chronic; Clinical; Clinical Trials; Combined Modality Therapy; Couples; Coupling; Diabetes Mellitus; Disease; Disease remission; Dose; Effector Cell; Endothelial Cells; Endothelium; Event; FDA approved; FOXO1A gene; Fibrosis; Flare; Functional disorder; Funding; Future; Gene Expression; General Population; Genes; Goals; Health; Heart Diseases; Histologic; Human; Hypertension; In Vitro; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Infiltrate; Innate Immune Response; Kidney; Kidney Diseases; Kidney Failure; Knowledge; Lead; Lupus; Lupus Nephritis; Mediating; Medicine; Methods; Modeling; Mus; Mycophenolate; Myocardial Infarction; NOS3 gene; Necrosis; Nephritis; Nitric Oxide; Onset of illness; Outcome; Oxidation-Reduction; Oxidative Stress; PDGFRB gene; Pathogenicity; Pathologic; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacodynamics; Platelet-Derived Growth Factor Receptor; Prevalence; Prevention; Production; Publishing; Reactive Oxygen Species; Remission Induction; Rodent; Serum; Signal Transduction; Spleen; Stimulus; Superoxides; Systemic Lupus Erythematosus; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Time; Tissue-Specific Gene Expression; Tissues; Translating; Vascular Diseases; Veterans; Withdrawal; Work; base; cytokine; effective therapy; effectiveness testing; endothelial dysfunction; experience; glomerular endothelium; hypertensive; immunoregulation; improved; improved outcome; in vivo; lupus-like; macrophage; macrovascular disease; monocyte; monomer; pharmacodynamic biomarker; podocyte; predicting response; prevent; primary endpoint; response biomarker; sepiapterin; single cell analysis; single-cell RNA sequencing; standard care; targeted treatment; tetrahydrobiopterin; therapeutic target; therapy development; transcriptome sequencing; translation to humans; treatment response

Systemic lupus erythematosus (SLE) is an immune complex-mediated disease in which clinical disease manifests after an innate immune response. Endothelial cells (EC) are conditional effector cells activated by inflammatory stimuli to produce intracellular reactive oxygen species (ROS) and signal for influx of inflammatory cells into tissue. We have published that endothelial nitric oxide synthase (eNOS) modulates inflammation in lupus nephritis (LN) and that uncoupling of EC eNOS (which reduces production of anti-inflammatory nitric oxide (NO) and increases production of pro-inflammatory ROS) is a potential therapeutic target in SLE. We hypothesize that sepiapterin, a compound that couples eNOS, will improve LN outcomes and vascular function in murine LN and work by modulating the PI3Kinase/Akt pathway to reduce expression of inflammation and fibrosis genes in mice and in cultured glomerular endothelial cells. We further hypothesize that the proposed approach may have off target effects, particularly in T cells, monocyte/macrophages, and podocytes. To address this overarching hypothesis, the following specific aims are proposed: Specific Aim 1) Determine the effect of sepiapterin in murine LN on clinical and histologic LN outcomes, large vessel endothelial function, and pharmacodynamic markers of treatment response. We hypothesize that sepiapterin will improve LN outcomes and vascular function in murine LN. To determine the effect of and optimal therapeutic indication for ECD targeted therapy, we will use the NZM2410 model of spontaneous LN and compare groups with and without sepiapterin treatment to determine effect on three different endpoints. 1) preventing/prolonging LN onset with sepiapterin monotherapy, 2) in mycophenolate-treated mice, improving/hastening induction of remission with sepiapterin as an adjunctive therapy, and 3) after inducing remission, prolonging/preventing LN flare with sepiapterin monotherapy after mycophenolate withdrawal. LN clinical and histologic parameters and large vessel endothelial function will be the primary endpoints. Biomarkers of systemic oxidative stress will serve as exploratory pharmacodynamic indicators. Specific Aim 2. Determine differential gene expression and phosphokinase signaling events, with a focus on the PI3K/Akt pathway, induced in human glomerular endothelial cells (HRGECs) by LN serum and modulated by sepiapterin. Based on our published and preliminary studies, we hypothesize that inflammatory gene expression and PDGF receptor/PI3K/Akt/FoxO1 signaling induced by LN flare serum can be modulated by sepiapterin in HRGECs. Knowledge of pathways activated by LN flare serum that both initiate redox-regulated pathways and that can be modulated by restoring coupling to eNOS will be important for rational development of therapies to target ECD in LN. We will culture HRGECs with serum from SLE patients with active nephritis or a cytokine mix and perturb signaling through the PDGFR, PI3K, and Akt to determine the effect on gene expression and NO and superoxide production. Specific Aim 3. Determine differential inflammatory and fibrosis gene expression and pathway enrichment in endothelial cells, infiltrating inflammatory cells, and podocytes from renal glomerular tissue and in T cells from the spleen in mice treated with increasing doses of sepiapterin. Based on our published and preliminary studies, we hypothesize that candidate PI3K-Akt signaling gene enrichment (both potentiated by and inducing of oxidative stress in a feed forward fashion) will be modulated by sepiapterin. NZM2410 mice will be treated with increasing doses of sepiapterin and renal cortical and spleen tissue analyzed by single cell RNA sequencing. The ultimate goal of our translational group is to target ECD to improve cardiovascular and renal outcomes in LN. Given the prevalence of cardiovascular disease in LN, we will study endothelium-dependent vascular function to inform early human studies of cardiovascular prevention. The aims proposed here will demonstrate the most effective treatment indications, pharmacodynamic markers of response, and alternate or adjunctive targets to reduce inflammatory ECD in LN and other diseases of endothelial dysfunction in Veterans.

系统性红斑狼疮（SLE）是一种免疫复合物介导的疾病， 会在先天免疫反应后出现内皮细胞（EC）是由内皮细胞激活的条件效应细胞。 炎性刺激产生细胞内活性氧（ROS）和炎性细胞内流的信号 细胞转化为组织。我们已经发表了内皮型一氧化氮合酶（eNOS）调节炎症， 狼疮肾炎（LN）和ECeNOS解偶联（其减少抗炎性一氧化氮的产生 (NO)并增加促炎性ROS的产生）是SLE的潜在治疗靶点。我们假设 sepiapterin是一种与eNOS结合的化合物， 并通过调节PI 3激酶/Akt通路来减少炎症和纤维化基因的表达， 小鼠和培养的肾小球内皮细胞。我们进一步假设，所提出的方法可能具有 脱靶效应，特别是在T细胞、单核细胞/巨噬细胞和足细胞中。为了解决这一总体问题， 具体目的1）确定sepiapterin在 小鼠LN对临床和组织学LN结局、大血管内皮功能以及 治疗反应的药效学标志物。我们假设sepiapterin将改善LN 结果和血管功能。目的：确定治疗的效果和最佳治疗适应症， ECD靶向治疗，我们将使用NZM 2410自发性LN模型，并比较 sepiapterin治疗以确定对三个不同终点的作用。1)预防/延长LN发作， sepiapterin单一疗法，2）在霉酚酸酯治疗的小鼠中，改善/加速诱导缓解， sepiapterin作为后续治疗，和3）诱导缓解后，延长/预防LN发作， 霉酚酸酯停药后的sepiapterin单药治疗。LN临床和组织学参数与大血管 内皮功能将是主要终点。全身性氧化应激的生物标志物将作为 探索性药效学指标。具体目标2。确定差异基因表达， 磷酸激酶信号传导事件，重点是PI 3 K/Akt通路，在人肾小球中诱导 内皮细胞（HRGECs）的LN血清和调节的sepiapterin。根据我们发布的和 初步研究，我们假设炎症基因表达和PDGF受体/PI 3 K/Akt/FoxO 1 LN爆发血清诱导的信号传导可通过HRGECs中的sepiapterin调节。途径知识 由LN爆发血清激活，启动氧化还原调节途径，并可以通过恢复来调节 与eNOS的偶联对于合理开发靶向LN中ECD的疗法将是重要的。我们将培养 HRGECs与来自活动性肾炎SLE患者的血清或细胞因子混合物，并通过 PDGFR、PI 3 K和Akt以确定对基因表达以及NO和超氧化物产生的影响。具体 目标3。确定不同的炎症和纤维化基因表达和途径富集， 内皮细胞、浸润性炎性细胞和肾小球组织中的足细胞以及T细胞中的足细胞 来自用递增剂量的sepiapterin处理的小鼠的脾的细胞。根据我们发布的和 初步研究中，我们假设候选PI 3 K-Akt信号转导基因富集（通过和 以前馈方式诱导氧化应激）将被sepiapterin调节。NZM 2410小鼠将 用递增剂量的Sepiapterin处理，通过单细胞RNA分析肾皮质和脾组织 测序我们翻译小组的最终目标是靶向ECD，以改善心血管和肾脏疾病。 结果在LN。鉴于LN患者心血管疾病的患病率，我们将研究内皮依赖性 血管功能，为心血管预防的早期人类研究提供信息。这里提出的目标将 证明最有效的治疗适应症、反应的药效学标志物和替代或 减少退伍军人LN和其他内皮功能障碍疾病中炎症ECD的靶点。