Novel Endothelial and Immunologic Mechanisms of Pathophysiology in Sickle Cell Disease

项目摘要 镰状细胞病于1910年首次在美国被发现，直到20世纪80年代才提出第一种治疗方法。 它是第一个确定的单基因致病性疾病，也是美国最常见的遗传性血液疾病。 当个体遗传两个突变的β珠蛋白基因时，每个基因来自父母。肾脏疾病被发现 在大量镰状细胞病（镰状细胞肾病）患者中，并可迅速进展至死亡。 肽内皮素1水平升高和免疫功能失调已被证明是主要的 在镰状细胞肾病的发展中起作用，然而，细胞来源为肽和免疫细胞类型 这种情况发生的原因是错误的。拟议研究的中心假设是，在 肾脏，镰状细胞病促进内皮特异性内皮素1表达和TH 17介导的损伤， 导致肾损伤、功能障碍和整体肾病。研究内皮源性 内皮素1，我们成功地建立了内皮特异性内皮素1基因敲除小鼠模型， 汤斯人源化镰状细胞小鼠背景。这些动物将被严格研究，以评估 肾损伤和功能障碍的标志物，并将进行研究，以确定 疾病进展。为了确定TH 17细胞在镰状细胞性肾病中的创新作用， 将进行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 进展和免疫细胞浸润。此外，使用已建立的过继转移实验，SCD 小鼠将具有来自在幼年时转移到其中的TH 17报告小鼠的体外极化的TH 17细胞， 评估对镰状细胞肾病的长期影响。最后，我们还将评估内皮细胞- 衍生的内皮素1在最近观察到SCD小鼠具有升高的肾内TH 17细胞中具有作用。 这两种创新的方法将寻求提供新的途径，以更好地靶向镰状细胞 肾病并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.