Ossoli Alice, Neufeld Edward B, Thacker Seth G, Vaisman Boris, Pryor Milton, Freeman Lita A, Brantner Christine A, Baranova Irina, Francone Nicolás O, Demosky Stephen J, Vitali Cecilia, Locatelli Monica, Abbate Mauro, Zoja Carlamaria, Franceschini Guido, Calabresi Laura, Remaley Alan T
Centro Grossi Paoletti, Dipartimento di Scienze Farmacologiche e Biomolecolari, Università degli Studi di Milano, Milano, Italy.
Lipoprotein Metabolism Section, Cardiovascular and Pulmonary Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
PLoS One. 2016 Feb 26;11(2):e0150083. doi: 10.1371/journal.pone.0150083. eCollection 2016.
Human familial lecithin:cholesterol acyltransferase (LCAT) deficiency (FLD) is characterized by low HDL, accumulation of an abnormal cholesterol-rich multilamellar particle called lipoprotein-X (LpX) in plasma, and renal disease. The aim of our study was to determine if LpX is nephrotoxic and to gain insight into the pathogenesis of FLD renal disease. We administered a synthetic LpX, nearly identical to endogenous LpX in its physical, chemical and biologic characteristics, to wild-type and Lcat-/- mice. Our in vitro and in vivo studies demonstrated an apoA-I and LCAT-dependent pathway for LpX conversion to HDL-like particles, which likely mediates normal plasma clearance of LpX. Plasma clearance of exogenous LpX was markedly delayed in Lcat-/- mice, which have low HDL, but only minimal amounts of endogenous LpX and do not spontaneously develop renal disease. Chronically administered exogenous LpX deposited in all renal glomerular cellular and matrical compartments of Lcat-/- mice, and induced proteinuria and nephrotoxic gene changes, as well as all of the hallmarks of FLD renal disease as assessed by histological, TEM, and SEM analyses. Extensive in vivo EM studies revealed LpX uptake by macropinocytosis into mouse glomerular endothelial cells, podocytes, and mesangial cells and delivery to lysosomes where it was degraded. Endocytosed LpX appeared to be degraded by both human podocyte and mesangial cell lysosomal PLA2 and induced podocyte secretion of pro-inflammatory IL-6 in vitro and renal Cxl10 expression in Lcat-/- mice. In conclusion, LpX is a nephrotoxic particle that in the absence of Lcat induces all of the histological and functional hallmarks of FLD and hence may serve as a biomarker for monitoring recombinant LCAT therapy. In addition, our studies suggest that LpX-induced loss of endothelial barrier function and release of cytokines by renal glomerular cells likely plays a role in the initiation and progression of FLD nephrosis.
人类家族性卵磷脂胆固醇酰基转移酶(LCAT)缺乏症(FLD)的特征是高密度脂蛋白(HDL)水平低、血浆中积聚一种称为脂蛋白-X(LpX)的异常富含胆固醇的多层颗粒以及肾脏疾病。我们研究的目的是确定LpX是否具有肾毒性,并深入了解FLD肾脏疾病的发病机制。我们将一种在物理、化学和生物学特性上与内源性LpX几乎相同的合成LpX给予野生型和Lcat-/-小鼠。我们的体外和体内研究表明,LpX转化为HDL样颗粒存在一条载脂蛋白A-I(apoA-I)和LCAT依赖性途径,这可能介导了LpX在血浆中的正常清除。外源性LpX在Lcat-/-小鼠中的血浆清除明显延迟,这些小鼠HDL水平低,但内源性LpX含量极少且不会自发发生肾脏疾病。长期给予外源性LpX会沉积在Lcat-/-小鼠的所有肾肾小球细胞和基质区室中,并诱导蛋白尿和肾毒性基因变化,以及通过组织学、透射电子显微镜(TEM)和扫描电子显微镜(SEM)分析评估的FLD肾脏疾病的所有特征。广泛的体内电子显微镜研究揭示,LpX通过巨吞饮作用被小鼠肾小球内皮细胞、足细胞和系膜细胞摄取,并输送到溶酶体中进行降解。内吞的LpX似乎被人足细胞和系膜细胞溶酶体磷脂酶A2降解,并在体外诱导足细胞分泌促炎白细胞介素-6(IL-6)以及在Lcat-/-小鼠中诱导肾脏Cxcl10表达。总之,LpX是一种肾毒性颗粒,在缺乏Lcat的情况下会诱发FLD的所有组织学和功能特征,因此可作为监测重组LCAT治疗的生物标志物。此外,我们的研究表明,LpX诱导的肾小球内皮屏障功能丧失和肾小球细胞释放细胞因子可能在FLD肾病的发生和发展中起作用。