Hyaluronidase 1 (Hyal-1) is an important enzyme involved in intracellular hyaluronic acid (HA) catabolism for performing various physiological functions, and its aberrant level is closely associated with many malignant diseases. Bioluminescence imaging is advantageous for monitoring Hyal-1 activity in vivo, but it remains challenging to design an available probe for differentiating Hyal-1 from other isoforms by a traditional strategy that covalently masks the firefly luciferase substrate. Herein, we, for the first time, present a noncovalently caging approach to construct a Hyal-1-specific bioluminogenic nanosensor by entrapping d-luciferin (d-Luc) inside the cholesterylamine-modified HA (CHA) nanoassembly to inhibit the bioluminescence production. When encountered with intracellular Hyal-1, CHA could be fully dissembled to liberate multiple copies of the loaded d-Luc, thereby emitting light by the luciferase-catalyzed bioluminescence reaction. Because of its cascade signal amplification feature, d-Luc@CHA displayed a remarkable “turn-on” response (248-fold) to 5 μg/mL Hyal-1 with a detection limit of 0.07 ng/mL. Importantly, bioluminescence imaging results validated that d-Luc@CHA could be competent for dynamically visualizing endogenous Hyal-1 changes in living cells and animals and possessed the capability of discriminating between normal and cancer cells, thus offering a promising toolbox to evaluate Hyal-1 roles in biological processes as well as to diagnose Hyal-1-related diseases.

中文翻译：

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非共价笼养萤火虫萤光素使体内的透明质酸酶1活性可放大的生物发光传感。

透明质酸酶1（Hyal-1）是参与细胞内透明质酸（HA）分解代谢的重要酶，具有多种生理功能，其异常水平与许多恶性疾病密切相关。生物发光成像对于监测体内的Hyal-1活性是有利的，但是通过共价掩盖萤火虫荧光素酶底物的传统策略，设计一种可用于将Hyal-1与其他亚型区分开的探针仍然具有挑战性。在此，我们，对于第一次，本非共价进行封闭而方法通过将药物包埋到构建HYAL-1特异性bioluminogenic纳米传感器d -luciferin（d-Luc）位于胆固醇胺修饰的HA（CHA）纳米组件内部，以抑制生物发光的产生。当遇到细胞内Hyal-1时，CHA可以被完全分解，以释放负载的d -Luc的多个副本，从而通过荧光素酶催化的生物发光反应发光。由于其级联信号放大功能，d -Luc @ CHA对5μg/ mL Hyal-1表现出显着的“开启”响应（248倍），检出限为0.07 ng / mL。重要的是，生物发光成像结果验证了d-Luc @ CHA可以动态观察活细胞和动物中内源性Hyal-1的变化，并具有区分正常细胞和癌细胞的能力，因此提供了一个有前途的工具箱，可评估Hyal-1在生物过程中的作用以及诊断与Hyal-1相关的疾病。