Identification of Cholesterol-Regulating Genes by Targeted RNAi Screening
Identification of Cholesterol-Regulating Genes by Targeted RNAi Screening
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DOI：
10.1016/j.cmet.2009.05.009
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发表时间：
2009-07-08
期刊：
影响因子：
29
通讯作者：
Runz, Heiko
中科院分区：
文献类型：
--
作者：
Bartz, Fabian;Kern, Luise;Runz, Heiko
血浆胆固醇水平升高被认为是心血管疾病发病率和死亡率过高的原因。血浆中的胆固醇受到细胞内胆固醇的严格控制。在这里，我们开发并应用了一种综合功能基因组学策略，可以系统地识别细胞胆固醇水平的调节因子。候选基因通过甾醇耗尽细胞的全基因组基因表达谱和系统的文献查询来鉴定。然后通过靶向siRNA敲低实验来测试这些基因在胆固醇调节中的作用，量化细胞胆固醇水平和低密度脂蛋白（LDL）摄取的效率。通过这种策略，20个基因被确定为细胞胆固醇稳态的功能调节因子。其中，我们将TMEM97描述为SREBP靶基因，该基因在胆固醇消耗条件下定位于内溶酶体/溶酶体室，并结合LDL胆固醇转运调节蛋白Niemann-Pick C1 （NPC1）。综上所述，TMEM97和其他因子有望进一步深入了解细胞如何控制胆固醇水平。
Elevated plasma cholesterol levels are considered responsible for excess cardiovascular morbidity and mortality. Cholesterol in plasma is tightly controlled by cholesterol within cells. Here, we developed and applied an integrative functional genomics strategy that allows systematic identification of regulators of cellular cholesterol levels. Candidate genes were identified by genome-wide gene-expression profiling of sterol-depleted cells and systematic literature queries. The role of these genes in cholesterol regulation was then tested by targeted siRNA knockdown experiments quantifying cellular cholesterol levels and the efficiency of low-density lipoprotein (LDL) uptake. With this strategy, 20 genes were identified as functional regulators of cellular cholesterol homeostasis. Of these, we describe TMEM97 as SREBP target gene that under sterol-depleted conditions localizes to endo-/lysosomal compartments and binds to LDL cholesterol transport-regulating protein Niemann-Pick C1 (NPC1). Taken together, TMEM97 and other factors described here are promising to yield further insights into how cells control cholesterol levels.