Tri-University Meson Facility, 4004 Wesbrook Mall, Vancouver, BC, Canada.
Proc Natl Acad Sci U S A. 2010 Jun 15;107(24):10842-7. doi: 10.1073/pnas.1003247107. Epub 2010 Jun 1.
Direct enzyme replacement therapy (ERT) has been introduced as a means to treat a number of rare, complex genetic conditions associated with lysosomal dysfunction. Gaucher disease was the first for which this therapy was applied and remains the prototypical example. Although ERT using recombinant lysosomal enzymes has been shown to be effective in altering the clinical course of Gaucher disease, Fabry disease, Hurler syndrome, Hunter syndrome, Maroteaux-Lamy syndrome, and Pompe disease, the recalcitrance of certain disease manifestations underscores important unanswered questions related to dosing regimes, tissue half-life of the recombinant enzyme and the ability of intravenously administered enzyme to reach critical sites of known disease pathology. We have developed an innovative method for tagging acid beta-glucocerebrosidase (GCase), the recombinant enzyme formulated in Cerezyme(R) used to treat Gaucher disease, using an (18)F-labeled substrate analogue that becomes trapped within the active site of the enzyme. Using micro-PET we show that the tissue distribution of injected enzyme can be imaged in a murine model and that the PET data correlate with tissue (18)F counts. Further we show that PET imaging readily monitors pharmacokinetic changes effected by receptor blocking. The ability to (18)F-label GCase to monitor the enzyme distribution and tissue half-life in vivo by PET provides a powerful research tool with an immediate clinical application to Gaucher disease and a clear path for application to other ERTs.
直接酶替代疗法 (ERT) 已被引入,作为治疗与溶酶体功能障碍相关的多种罕见、复杂遗传疾病的一种手段。戈谢病是第一种应用这种治疗方法的疾病,也是典型的例子。尽管使用重组溶酶体酶的 ERT 已被证明可以有效改变戈谢病、法布里病、亨特综合征、马罗佐-拉米综合征和庞贝病的临床病程,但某些疾病表现的顽固性突出了与剂量方案、重组酶的组织半衰期以及静脉内给予的酶到达已知疾病病理学关键部位的能力相关的重要未解决问题。我们开发了一种创新的方法,使用(18)F 标记的底物类似物对用于治疗戈谢病的重组酶酸性β-葡糖苷脑苷脂酶 (GCase) 进行标记,该类似物可被捕获在酶的活性部位内。使用 micro-PET,我们表明可以在小鼠模型中对注射酶的组织分布进行成像,并且 PET 数据与组织(18)F 计数相关。此外,我们表明 PET 成像可以轻松监测受体阻断引起的药代动力学变化。通过 PET 用(18)F 标记 GCase 来监测体内酶分布和组织半衰期的能力提供了一种强大的研究工具,具有立即应用于戈谢病的临床应用和明确的应用于其他 ERT 的途径。