Rosenbaum Anton I, Rujoi Madalina, Huang Amy Y, Du Hong, Grabowski Gregory A, Maxfield Frederick R
Department of Biochemistry, Weill Cornell Medical College, 1300 York Avenue, New York, NY 10065, USA.
Biochim Biophys Acta. 2009 Dec;1791(12):1155-65. doi: 10.1016/j.bbalip.2009.08.005. Epub 2009 Aug 20.
Niemann-Pick C disease (NPC) is a lysosomal storage disorder causing abnormal accumulation of unesterified free cholesterol in lysosomal storage organelles. High content phenotypic microscopy chemical screens in both human and hamster NPC-deficient cells have identified several compounds that partially revert the NPC phenotype. Cell biological and biochemical studies show that several of these molecules inhibit lysosomal acid lipase, the enzyme that hydrolyzes LDL-derived triacylglycerol and cholesteryl esters. The effects of reduced lysosomal acid lipase activity in lowering cholesterol accumulation in NPC mutant cells were verified by RNAi-mediated knockdown of lysosomal acid lipase in NPC1-deficient human fibroblasts. This work demonstrates the utility of phenotypic cellular screens as a means to identify molecular targets for altering a complex process such as intracellular cholesterol trafficking and metabolism.
尼曼-匹克C型病(NPC)是一种溶酶体贮积症,会导致溶酶体贮积细胞器中未酯化游离胆固醇异常蓄积。在人类和仓鼠NPC缺陷细胞中进行的高内涵表型显微镜化学筛选已鉴定出几种可部分逆转NPC表型的化合物。细胞生物学和生化研究表明,其中几种分子可抑制溶酶体酸性脂肪酶,该酶可水解低密度脂蛋白衍生的三酰甘油和胆固醇酯。通过RNAi介导的NPC1缺陷型人成纤维细胞中溶酶体酸性脂肪酶的敲低,证实了溶酶体酸性脂肪酶活性降低对降低NPC突变细胞中胆固醇蓄积的作用。这项工作证明了表型细胞筛选作为一种识别改变细胞内胆固醇转运和代谢等复杂过程的分子靶点的手段的实用性。