Division of Translational Medicine and Human Genetics, Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.
Office of Institutional Research & Analysis, University of Pennsylvania, Philadelphia, PA, USA.
J Alzheimers Dis. 2020;74(2):563-578. doi: 10.3233/JAD-191246.
Basic research has implicated intracellular cholesterol in neurons, microglia, and astrocytes in the pathogenesis of Alzheimer's disease (AD), but there is presently no assay to access intracellular cholesterol in neural cells in living people in the context of AD.
To devise and characterize an assay that can access intracellular cholesterol and cholesterol efflux in neural cells in living subjects.
We modified the protocol for high-density lipoprotein cholesterol efflux capacity (CEC) from macrophages, a biomarker that accesses cholesterol in macrophages in atherosclerosis. To measure cerebrospinal fluid (CSF) CECs from neurons, microglia, and astrocytes, CSF was exposed to, correspondingly, neuronal, microglial, and astrocytic cholesterol source cells. Human neuroblastoma SH-SY5Y, mouse microglial N9, and human astroglial A172 cells were used as the cholesterol source cells. CSF samples were screened for contamination with blood. CSF CECs were measured in a small cohort of 22 individuals.
CSF CECs from neurons, microglia, and astrocytes were moderately to moderately strongly correlated with CSF concentrations of cholesterol, apolipoprotein A-I, apolipoprotein E, and clusterin (Pearson's r = 0.53-0.86), were in poor agreement with one another regarding CEC of the CSF samples (Lin's concordance coefficient rc = 0.71-0.76), and were best predicted by models consisting of, correspondingly, CSF phospholipid (R2 = 0.87, p < 0.0001), CSF apolipoprotein A-I and clusterin (R2 = 0.90, p < 0.0001), and CSF clusterin (R2 = 0.62, p = 0.0005).
Characteristics of the CSF CEC metrics suggest a potential for independent association with AD and provision of fresh insight into the role of cholesterol in AD pathogenesis.
基础研究表明，阿尔茨海默病（AD）患者的神经元、小胶质细胞和星形胶质细胞内的胆固醇与疾病的发病机制有关，但目前尚无检测方法可以在 AD 患者活体中检测神经细胞内的细胞内胆固醇。
设计并描述一种可以检测活体受试者神经细胞内细胞内胆固醇和胆固醇外排的检测方法。
我们修改了高密度脂蛋白胆固醇流出能力（CEC）的测定方案，该方案来自动脉粥样硬化中巨噬细胞的生物标志物，可以测定巨噬细胞内的胆固醇。为了测量神经元、小胶质细胞和星形胶质细胞的脑脊液（CSF）CEC，将 CSF 暴露于相应的神经元、小胶质细胞和星形胶质细胞胆固醇源细胞。使用人神经母细胞瘤 SH-SY5Y、小鼠小胶质细胞 N9 和人星形胶质细胞 A172 作为胆固醇源细胞。筛选 CSF 样本是否存在血液污染。在一个由 22 人组成的小队列中测量 CSF CEC。
神经元、小胶质细胞和星形胶质细胞的 CSF CEC 与 CSF 胆固醇、载脂蛋白 A-I、载脂蛋白 E 和载脂蛋白 E 浓度中度至中度高度相关（皮尔逊 r=0.53-0.86）， CSF CEC 之间的一致性较差（林氏一致性系数 rc=0.71-0.76），最能由相应的 CSF 磷脂（R2=0.87，p<0.0001）、CSF 载脂蛋白 A-I 和载脂蛋白 E（R2=0.90，p<0.0001）和 CSF 载脂蛋白 E（R2=0.62，p=0.0005）组成的模型来预测。
CSF CEC 指标的特点表明其与 AD 有潜在的独立关联，并为胆固醇在 AD 发病机制中的作用提供了新的见解。
