Novel Endothelial and Immunologic Mechanisms of Pathophysiology in Sickle Cell Disease

项目摘要 镰状细胞病最早于1910年在美国发现，直到20世纪80年代S才提出第一种治疗方法。 这是第一种被确认的单基因致病疾病，也是美国最常见的遗传性血液疾病。 当个体遗传了两个突变的β珠蛋白基因时，就会出现这种情况，父母各一个。发现肾脏疾病 在大量患有镰状细胞病(镰状细胞肾病)的患者中，可迅速进展至死亡。 研究表明，多肽ET-1水平升高和免疫功能失调是主要原因 在镰状细胞肾病的发生发展中的作用，然而，多肽和免疫细胞类型的细胞来源 对于这种情况发生的原因，人们很难理解。拟议研究的中心假设是，在 肾脏，镰状细胞疾病促进内皮特异性内皮素1的表达和TH17介导的损伤， 导致肾脏损伤、功能障碍和整体肾病。研究血管内皮细胞衍生细胞的作用 ，我们成功地建立了一种内皮特异性内皮素1基因敲除小鼠模型。 汤斯人源化镰状细胞小鼠背景。将对这些动物进行严格的研究，以评估 肾脏损伤和功能障碍的标志物，并将进行研究，以确定肾脏损伤和功能障碍的病理生理特征 疾病的发展。确定TH17细胞在镰状细胞肾病中的创新作用，人源化小鼠 将接受流式细胞术和组织病理学分析，以确定疾病之间的时间关系 进展和免疫细胞的渗透。此外，利用已建立的采用迁移实验，SCD 小鼠将在体外将TH17报告鼠的极化TH17细胞转移到小鼠体内，以 评估远期对镰状细胞性肾病的贡献。最后，我们还将评估内皮细胞是否- 在最近观察到SCD小鼠肾内TH17细胞升高的过程中，衍生的内皮素1起了作用。 这两种创新方法将寻求提供更好地靶向镰状细胞的新途径 治疗肾病，最终改善镰状细胞病患者的预后。

Project Summary Sickle cell disease was first identified in the US in 1910 and the first treatment was not proposed until the 1980’s. It is the first identified single gene causative disease and is the most common inherited blood disorder in the US. It presents when individuals inherit two mutant beta globin genes, one from each parent. Kidney disease is found in a large number of patients with sickle cell disease (sickle cell nephropathy) and can progress to death rapidly. Elevated levels of the peptide endothelin 1 and dysregulated immune function have been shown to play a major role in the development of sickle cell nephropathy, However, the cell source for the peptide and immune cell type by which this occurs is poorly misunderstood. The central hypothesis of the proposed studies is that, within the kidney, sickle cell disease promotes endothelial-specific endothelin 1 expression and TH17-mediated damage, leading to renal injury, dysfunction and overall nephropathy. To study the contribution of endothelial-derived endothelin 1, we successfully developed an endothelial-specific endothelin 1 knockout mouse model on the Townes humanized sickle cell mouse background. These animals will be studied rigorously in order to evaluate markers of renal injury and dysfunction, and will undergo studies to determine pathophysiological hallmarks of disease progression. To determine the innovative role of TH17 cells in sickle cell nephropathy, humanized mice will undergo flow cytometric and histopathologic analysis to determine temporal relationship between disease progression and immune cell infiltration. Furthermore, using established adoptive transfer experiments, SCD mice will have in vitro polarized TH17 cells from TH17 reporter mice transferred into them at a young age to evaluate the contribution long-term to sickle cell nephropathy. Finally, we will also evaluate whether endothelial- derived endothelin 1 has a role in the recent observation that SCD mice have elevated intrarenal TH17 cells. These two innovative approaches will seek to provide novel pathways by which to better target sickle cell nephropathy and ultimately improve outcomes for patients with sickle cell disease.