Rappaport Jeff, Garnacho Carmen, Muro Silvia
Fischell Department of Bioengineering, University of Maryland , College Park, Maryland 20742-4450, United States.
Mol Pharm. 2014 Aug 4;11(8):2887-95. doi: 10.1021/mp500241y. Epub 2014 Jun 26.
Drugs often use endocytosis to achieve intracellular delivery, either by passive uptake from the extracellular fluid or by active targeting of cell surface features such as endocytic receptors. An example is enzyme replacement therapy, a clinically practiced treatment for several lysosomal storage diseases where glycosylated recombinant enzymes naturally target the mannose-6-phosphate receptor and are internalized by clathrin mediated endocytosis (CME). However, lysosomal substrate accumulation, a hallmark of these diseases, has been indirectly linked to aberrant endocytic activity. These effects are poorly understood, creating an obstacle to therapeutic efficiency. Here we explored endocytic activity in fibroblasts from patients with type A Niemann-Pick disease, a lysosomal storage disease characterized by acid sphingomyelinase (ASM) deficiency. The uptake of fluid phase markers and clathrin-associated ligands, formation of endocytic structures, and recruitment of intracellular clathrin to ligand binding sites were all altered, demonstrating aberrant CME in these cells. Model polymer nanocarriers targeted to intercellular adhesion molecule-1 (ICAM-1), which are internalized by a clathrin-independent route, enhanced the intracellular delivery of recombinant ASM more than 10-fold compared to free enzyme. This strategy reduced substrate accumulation and restored clathrin endocytic activity to wild-type levels. There appears to be a relationship between lysosomal storage and diminished CME, and bypassing this pathway by targeting ICAM-1 may enhance future therapies for lysosomal storage diseases.
药物通常利用内吞作用实现细胞内递送,其方式要么是从细胞外液被动摄取,要么是主动靶向细胞表面特征,如内吞受体。酶替代疗法就是一个例子,这是一种临床上用于治疗多种溶酶体贮积病的方法,其中糖基化重组酶会自然靶向甘露糖-6-磷酸受体,并通过网格蛋白介导的内吞作用(CME)被内化。然而,这些疾病的一个标志——溶酶体底物积累,已被间接证明与异常的内吞活性有关。人们对这些影响了解甚少,这给治疗效率带来了障碍。在这里,我们研究了A型尼曼-匹克病患者成纤维细胞中的内吞活性,这是一种以酸性鞘磷脂酶(ASM)缺乏为特征的溶酶体贮积病。液相标记物和网格蛋白相关配体的摄取、内吞结构的形成以及细胞内网格蛋白向配体结合位点的募集均发生了改变,表明这些细胞中存在异常的CME。靶向细胞间黏附分子-1(ICAM-1)的模型聚合物纳米载体通过不依赖网格蛋白的途径被内化,与游离酶相比,其重组ASM的细胞内递送增强了10倍以上。该策略减少了底物积累,并将网格蛋白内吞活性恢复到野生型水平。溶酶体贮积与CME减弱之间似乎存在关联,通过靶向ICAM-1绕过这一途径可能会增强未来对溶酶体贮积病的治疗效果。