Wolman disease/cholesteryl ester storage disease: efficacy of plant-produced human lysosomal acid lipase in mice
Wolman disease/cholesteryl ester storage disease: efficacy of plant-produced human lysosomal acid lipase in mice
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DOI：
10.1194/jlr.m700482-jlr200
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发表时间：
2008-08-01
影响因子：
6.5
通讯作者：
Grabowski, Gregory A.
中科院分区：
文献类型：
--
作者：
Du, Hong;Cameron, Terri L.;Grabowski, Gregory A.
溶酶体酸性脂肪酶（LAL）是溶酶体中水解甘油三酯（tg）和胆固醇酯（CEs）的必需酶。LAL基因突变可导致沃尔曼病（WD）和胆固醇酯沉积病（CESD）。lal -null （lal(-/-)）小鼠模型与人类WD/CESD相似，在多个器官中储存了ce和tg。利用GENEWARE(R)表达系统（g - hal）在烟叶中表达人LAL （hal）。纯化的g - hal显示甘露糖受体依赖摄取到巨噬细胞系（J774E）。腹腔注射g - hal在60 min时血浆、240 min时肝脏和脾脏的活性达到峰值。t（1/2）值为：与90分钟（血浆）相近，与14小时（肝脏）相近，与32小时（脾脏）相近，与150小时（肝脏）相近，与200小时（脾脏）相近。向lal（-/-）小鼠注射10次g - hal（每3天一次），肝脏颜色正常化，肝脏胆固醇和TG含量降低，肝脏、脾脏和肠绒毛泡沫巨噬细胞减少。所有注射了lal（-/-）的小鼠都产生了抗hal蛋白抗体，但没有发生不良事件。这些研究表明，利用植物表达的重组hal酶治疗人WD/CESD的可行性，对其他溶酶体贮积性疾病具有普遍意义。
Lysosomal acid lipase (LAL) is an essential enzyme that hydrolyzes triglycerides (TGs) and cholesteryl esters (CEs) in lysosomes. Genetic LAL mutations lead to Wolman disease (WD) and cholesteryl ester storage disease (CESD). An LAL-null (lal(-/-)) mouse model resembles human WD/CESD with storage of CEs and TGs in multiple organs. Human LAL (hLAL) was expressed in Nicotiana benthamiana using the GENEWARE(R) expression system (G-hLAL). Purified G-hLAL showed mannose receptor-dependent uptake into macrophage cell lines (J774E). Intraperitoneal injection of G-hLAL produced peak activities in plasma at 60 min and in the liver and spleen at 240 min. The t(1/2) values were: similar to 90 min (plasma), similar to 14 h (liver), and similar to 32 h (spleen), with return to baseline by similar to 150 h in liver and similar to 200 h in spleen. Ten injections of G-hLAL (every 3 days) into lal(-/-) mice produced normalization of hepatic color, decreases in hepatic cholesterol and TG contents, and diminished foamy macrophages in liver, spleen, and intestinal villi. All injected lal(-/-) mice developed anti-hLAL protein antibodies, but suffered no adverse events. These studies demonstrate the feasibility of using plant-expressed, recombinant hLAL for the enzyme therapy of human WD/CESD with general implications for other lysosomal storage diseases.