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，临床疾病 在先天免疫反应后显现。内皮细胞 (EC) 是由以下物质激活的条件效应细胞 炎症刺激产生细胞内活性氧（ROS）和炎症流入信号 细胞转化为组织。我们已经发表了内皮一氧化氮合酶 (eNOS) 调节炎症 狼疮性肾炎 (LN) 和 EC eNOS 解偶联（减少抗炎一氧化氮的产生） (NO) 并增加促炎性 ROS 的产生）是 SLE 的潜在治疗靶点。我们假设 墨蝶呤（一种与 eNOS 偶联的化合物）将改善小鼠 LN 的 LN 结局和血管功能 并通过调节 PI3Kinase/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。为了解决这个总体问题 假设，提出以下具体目标： 具体目标 1) 确定墨蝶呤在 小鼠 LN 对临床和组织学 LN 结果、大血管内皮功能和 治疗反应的药效学标志物。我们假设墨蝶呤会改善 LN 小鼠 LN 的结果和血管功能。确定效果和最佳治疗适应症 ECD靶向治疗，我们将使用自发性LN的NZM2410模型并比较有和没有的组 墨蝶呤治疗以确定对三个不同终点的影响。 1) 预防/延长 LN 发作 墨蝶呤单一疗法，2) 在霉酚酸酯治疗的小鼠中，改善/加速诱导缓解 墨蝶呤作为辅助治疗，以及 3) 在诱导缓解后，延长/预防 LN 发作 麦考酚酯停药后，墨蝶呤单药治疗。 LN 临床和组织学参数以及大血管 内皮功能将是主要终点。全身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. 确定差异基因表达和 在人肾小球中诱导的磷酸激酶信号传导事件，重点是 PI3K/Akt 通路 由 LN 血清和墨蝶呤调节的内皮细胞 (HRGEC)。根据我们发布的和 初步研究，我们假设炎症基因表达与PDGF受体/PI3K/Akt/FoxO1 LN 耀斑血清诱导的信号传导可通过 HRGEC 中的墨蝶呤进行调节。途径知识 由 LN 耀斑血清激活，既启动氧化还原调节途径，又可以通过恢复来调节 与 eNOS 偶联对于合理开发针对 LN 中 ECD 的疗法非常重要。我们将文化 HRGECs 含有活动性肾炎 SLE 患者的血清或细胞因子混合物，并通过 PDGFR、PI3K 和 Akt 以确定对基因表达以及 NO 和超氧化物产生的影响。具体的 目标 3. 确定不同的炎症和纤维化基因表达以及通路富集 肾小球组织和 T 中的内皮细胞、浸润性炎症细胞和足细胞 来自接受增加剂量的墨蝶呤治疗的小鼠的脾脏细胞。根据我们发布的和 初步研究中，我们假设候选 PI3K-Akt 信号基因富集（均由 和 以前馈方式诱导氧化应激）将受到墨蝶呤的调节。 NZM2410 小鼠将 用增加剂量的墨蝶呤处理，并通过单细胞 RNA 分析肾皮质和脾组织 测序。我们转化小组的最终目标是针对 ECD 来改善心血管和肾脏疾病 LN 中的结果。鉴于 LN 中心血管疾病的患病率，我们将研究内皮依赖性 血管功能为早期人类心血管预防研究提供信息。这里提出的目标将 证明最有效的治疗适应症、反应的药效标志物以及替代或 减少 LN 炎症性 ECD 和退伍军人其他内皮功能障碍疾病的辅助目标。