Renal endothelium and the development of chronic kidney disease in sickle cell disorders

镰状细胞病中的肾内皮和慢性肾病的发展

• 批准号: 10659575
• 负责人: Samit Ghosh
• 金额: $ 65.15万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10659575
• 关键词: Abnormal Red Blood Cell; Acceleration; Adult; Affect; Albuminuria; Anabolism; Anemia; Binding; Biological Markers; Blood Vessels; Bone Marrow Transplantation; Cell Adhesion Molecules; Cell membrane; Cells; Characteristics; Childhood; Chronic; Chronic Kidney Failure; Clinical; Clinical Data; DNA; Data; Deterioration; Development; Disease Progression; Dose; Endothelial Cells; Endothelium; Event; Flow Cytometry; Foundations; Frequencies; Genotype; Glomerular Filtration Rate; Health; Heme; Hemin; Hemolysis; Histone Acetylation; Histone Deacetylase Inhibitor; Human; Impairment; In Vitro; Individual; Infusion procedures; Injections; Intravenous; Kidney; Lead; Lysine; Measures; Microalbuminuria; Modeling; Molecular; Morbidity - disease rate; Mus; Mutant Strains Mice; NF-kappa B; Organ; Organ failure; P-Selectin; PI3 gene; Pathogenesis; Patients; Peptide Hydrolases; Permeability; Phenocopy; Pilot Projects; Plasma; Predisposition; Progressive Disease; Prolastin; Prophylactic treatment; Proteinase 3; Proteins; Receptor Gene; Renal function; Research; Risk; Sampling; Selectins; Severities; Sickle Cell Anemia; Single Nucleotide Polymorphism; Specificity; Testing; Transgenic Organisms; Tube; Ultrasonography; Variant; Vascular Endothelium; Vascular Permeabilities; activated protein C receptor; age related; aged; biomarker discovery; cohort; disorder risk; drug discovery; endothelial dysfunction; epidemiology study; experimental study; extracellular vesicles; genetic variant; kidney vascular structure; mortality; mouse model; neutrophil; novel marker; overexpression; premature; prevent; receptor expression; sickling; targeted treatment; therapeutic target; ultra high resolution

PROJECT SUMMARY/ABSTRACT Chronic kidney disease (CKD), widespread among individuals with sickle cell disease (SCD), is a significant contributor to morbidity and early mortality. The progressive deterioration of renal health in SCD is associated with chronic intravascular hemolysis. However, the mechanistic approach to interpreting the hemolysis-driven pathogenesis of CKD in SCD is limited and therefore targeted therapies based on mechanistic models are lacking. Progressive loss of renal function, represented by increased albuminuria and reduced glomerular filtration rate (GFR), is associated with microvascular congestion in humans and mice with SCD. We postulated that circulating cell-free heme, a product of hemolysis, may instigate persistent endothelial damage that ensues CKD development. Endothelial protein C receptor (EPCR) maintains endothelial barrier integrity. We discovered an age-dependent as well as heme-induced loss of EPCR from renal microvascular endothelium in SCD (SS) mice. Concomitantly, the cleaved soluble form of EPCR (sEPCR) was elevated in the plasma associated with albuminuria. Using super-resolution ultrasound imaging, we found that younger SS mice challenged with the repetitive infusion of minimal doses of heme and the older SS mice without heme challenge incur substantial renal microvascular rarefaction. Heme induces endothelial P-selectin that promotes vascular congestion. Our pilot data show that reduced EPCR expression is associated with increased P-selectin on renal microvascular endothelium in the SS mice. Proteinase 3 (PR3), expressed on neutrophils, can degrade EPCR. In our repeated heme-challenged younger SS mice compared to vehicle-injected mice, we noticed increased expression of PR3 in plasma as well as in accumulated neutrophils from the kidneys. Moreover, we found that heme induces acetylation of histone 4 Lys 16 residues (H4K16ac) which are known to promote the biosynthesis of PR3. Based on these preliminary data, we hypothesize that heme-induced P-selectin and PR3 concurrently result in the loss of EPCR leading to endothelial disintegration, microvascular congestion, and CKD development in SCD. We will test this hypothesis with three specific aims that integrate experiments with human renal endothelial cells and neutrophils in vitro, murine models, and clinical samples from multiple cohorts of SCD patients. Aim 1 will determine that repeated low-dose heme insults promote renal endothelial P-selectin that suppresses EPCR leading to vascular damage and development of CKD in mice with SCD. Aim 2 will test whether heme- activated PR3 facilitates EPCR cleavage and expedite CKD in SCD. Aim 3 will determine whether sEPCR and specific genetic variants of EPCR can indicate the risk of CKD in SCD patients.

项目概要/摘要 慢性肾病 (CKD) 在镰状细胞病 (SCD) 患者中广泛存在，是一种重要的疾病 导致发病率和早期死亡率。 SCD 患者肾脏健康状况的逐渐恶化与 伴有慢性血管内溶血。然而，解释溶血驱动的机械方法 SCD 中 CKD 的发病机制有限，因此基于机制模型的靶向治疗是 缺乏。肾功能进行性丧失，表现为蛋白尿增加和肾小球减少 滤过率 (GFR) 与患有 SCD 的人类和小鼠的微血管充血有关。我们假设 循环中的无细胞血红素是溶血的产物，可能会引起持续的内皮损伤 CKD 发展。内皮蛋白 C 受体 (EPCR) 维持内皮屏障的完整性。我们发现 SCD (SS) 患者肾微血管内皮 EPCR 的年龄依赖性以及血红素诱导的损失 老鼠。与此同时，血浆中 EPCR 的裂解可溶形式 (sEPCR) 升高，与 蛋白尿。使用超分辨率超声成像，我们发现年轻的 SS 小鼠面临着 重复输注最小剂量的血红素和未进行血红素攻击的老年 SS 小鼠会产生大量的 肾微血管稀疏。血红素诱导内皮 P-选择素，促进血管充血。我们的 试验数据表明，EPCR 表达减少与肾微血管上 P-选择素增加相关 SS 小鼠的内皮细胞。中性粒细胞上表达的蛋白酶 3 (PR3) 可以降解 EPCR。在我们的反复 与注射载体的小鼠相比，受到血红素挑战的年轻 SS 小鼠，我们注意到 PR3 的表达增加 血浆以及肾脏中积累的中性粒细胞。此外，我们发现血红素诱导 组蛋白 4 Lys 16 残基 (H4K16ac) 的乙酰化已知可促进 PR3 的生物合成。基于 根据这些初步数据，我们假设血红素诱导的 P-选择素和 PR3 同时导致损失 EPCR 导致 SCD 中的内皮崩解、微血管充血和 CKD 发展。我们将 通过三个具体目标来检验这一假设，这些目标整合了人类肾内皮细胞的实验和 体外中性粒细胞、小鼠模型和来自多个 SCD 患者队列的临床样本。 目标 1 将确定重复的低剂量血红素损伤会促进肾内皮 P-选择素的抑制 EPCR 导致 SCD 小鼠血管损伤和 CKD 发展。目标 2 将测试血红素是否- 激活的 PR3 促进 EPCR 裂解并加速 SCD 中的 CKD。目标 3 将确定 sEPCR 和 EPCR 的特定基因变异可以提示 SCD 患者罹患 CKD 的风险。