Department of Chemistry, University of Basel, Klingelbergstrasse 80, CH-4056, Basel, Switzerland.
Department of Pharmaceutical Sciences, Division of Pharmaceutical Technology, University of Basel, Klingelbergstrasse 50, CH-4056, Basel, Switzerland.
Nat Commun. 2018 Mar 19;9(1):1127. doi: 10.1038/s41467-018-03560-x.
Despite tremendous efforts to develop stimuli-responsive enzyme delivery systems, their efficacy has been mostly limited to in vitro applications. Here we introduce, by using an approach of combining biomolecules with artificial compartments, a biomimetic strategy to create artificial organelles (AOs) as cellular implants, with endogenous stimuli-triggered enzymatic activity. AOs are produced by inserting protein gates in the membrane of polymersomes containing horseradish peroxidase enzymes selected as a model for natures own enzymes involved in the redox homoeostasis. The inserted protein gates are engineered by attaching molecular caps to genetically modified channel porins in order to induce redox-responsive control of the molecular flow through the membrane. AOs preserve their structure and are activated by intracellular glutathione levels in vitro. Importantly, our biomimetic AOs are functional in vivo in zebrafish embryos, which demonstrates the feasibility of using AOs as cellular implants in living organisms. This opens new perspectives for patient-oriented protein therapy.
尽管人们为开发响应性酶递药系统付出了巨大努力，但它们的疗效大多仅限于体外应用。在这里，我们通过将生物分子与人工隔室相结合的方法，引入了一种仿生策略，以创建作为细胞植入物的人工细胞器 (AO)，其具有内源性刺激触发的酶活性。AO 是通过在含有辣根过氧化物酶的聚合物囊泡的膜中插入蛋白门来制备的，辣根过氧化物酶酶被选为参与氧化还原动态平衡的天然酶的模型。插入的蛋白门是通过将分子帽附着到基因修饰的通道孔蛋白上来设计的，以诱导通过膜的分子流的氧化还原响应控制。AO 保留其结构，并在体外通过细胞内谷胱甘肽水平激活。重要的是，我们的仿生 AO 在斑马鱼胚胎中具有体内功能，这证明了将 AO 用作活体生物中的细胞植入物的可行性。这为面向患者的蛋白质治疗开辟了新的前景。
