Histone deacetylase inhibitors correct the cholesterol storage defect in most Niemann-Pick C1 mutant cells
Histone deacetylase inhibitors correct the cholesterol storage defect in most Niemann-Pick C1 mutant cells
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DOI：
10.1194/jlr.m072140
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发表时间：
2017-04-01
影响因子：
6.5
通讯作者：
Maxfield, Frederick R.
中科院分区：
文献类型：
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作者：
Pipalia, Nina H.;Subramanian, Kanagaraj;Maxfield, Frederick R.
尼曼-皮克C（NPC）病是一种常染色体隐性遗传疾病，导致胆固醇和其他脂质在晚期内体和溶酶体中过度储存。绝大多数NPC疾病是由NPC 1突变引起的，NPC 1是一种在晚期内体中起作用的大型多位膜蛋白。NPC 1基因存在多种疾病相关突变，多数患者为复合杂合子。最常见的突变，NPC 1（I1061 T），已被证明会导致内质网相关的NPC 1蛋白降解。用组蛋白脱乙酰酶抑制剂（HDACis）伏立诺他或帕比司他处理患者来源的NPC 1（I1061 T）成纤维细胞增加突变NPC 1蛋白的表达并导致胆固醇储存的校正。在这里，我们表明，其他几种人NPC 1突变成纤维细胞系也可以被伏立诺他或帕比司他纠正，并且伏立诺他治疗延长了NPC 1（I1061 T）蛋白的寿命。为了测试HDACi对大量NPC 1突变体的影响，我们设计了U2 OS细胞系以通过shRNA抑制NPC 1表达，然后用60种不同的NPC 1突变体构建体瞬时转染这些细胞。突变体NPC 1没有显著降低胆固醇积累，但当用伏立诺他或帕比司他处理时，约85%的突变体显示出降低的胆固醇积累。
Niemann-Pick C (NPC) disease is an autosomal recessive disorder that leads to excessive storage of cholesterol and other lipids in late endosomes and lysosomes. The large majority of NPC disease is caused by mutations in NPC1, a large polytopic membrane protein that functions in late endosomes. There are many disease-associated mutations in NPC1, and most patients are compound heterozygotes. The most common mutation, NPC1(I1061T), has been shown to cause endoplasmic reticulum-associated degradation of the NPC1 protein. Treatment of patient-derived NPC1(I1061T) fibroblasts with histone deacetylase inhibitors (HDACis) vorinostat or panobinostat increases expression of the mutant NPC1 protein and leads to correction of the cholesterol storage. Here, we show that several other human NPC1 mutant fibroblast cell lines can also be corrected by vorinostat or panobinostat and that treatment with vorinostat extends the lifetime of the NPC1(I1061T) protein. To test effects of HDACi on a large number of NPC1 mutants, we engineered a U2OS cell line to suppress NPC1 expression by shRNA and then transiently transfected these cells with 60 different NPC1 mutant constructs. The mutant NPC1 did not significantly reduce cholesterol accumulation, but approximately 85% of the mutants showed reduced cholesterol accumulation when treated with vorinostat or panobinostat.