5-Aminolevulinic acid-based photodynamic therapy heavily depends on the biological transformation efficiency of 5-aminolevulinic acid to protoporphyrin IX, while the lack of an effective delivery system and imaging navigation are major hurdles in improving the accumulation of protoporphyrin IX and optimizing therapeutic parameters. Herein, we leverage a synthetic biology approach to construct a transdermal theranostic microneedle patch integrated with 5-aminolevulinic acid and catalase co-loaded tumor acidity-responsive copper-doped calcium phosphate nanoparticles for efficient 5-aminolevulinic acid-based photodynamic therapy by maximizing the enrichment of intratumoral protoporphyrin IX. We show that continuous oxygen generation by catalase in vivo reverses tumor hypoxia, enhances protoporphyrin IX accumulation by blocking protoporphyrin IX efflux (downregulating hypoxia-inducible factor-1α and ferrochelatase) and upregulates protoporphyrin IX biosynthesis (providing exogenous 5-aminolevulinic acid and upregulating ALA-synthetase). In vivo fluorescence/photoacoustic duplex imaging can monitor intratumoral oxygen saturation and protoporphyrin IX metabolic kinetics simultaneously. This approach thus facilitates the optimization of therapeutic parameters for different cancers to realize Ca²⁺/Cu²⁺-interferences-enhanced repeatable photodynamic therapy, making this theranostic patch promising for clinical practice.

基于5-氨基乙酰丙酸的光动力疗法很大程度上取决于5-氨基乙酰丙酸向原卟啉IX的生物转化效率，而缺乏有效的递送系统和成像导航是提高原卟啉IX的积累和优化治疗参数的主要障碍。在此，我们利用合成生物学方法构建了一种透皮治疗微针贴片，该微针贴片集成了 5-氨基乙酰丙酸和过氧化氢酶共负载的肿瘤酸度响应型铜掺杂磷酸钙纳米颗粒，通过最大化富集来实现有效的基于 5-氨基乙酰丙酸的光动力治疗瘤内原卟啉 IX。我们发现过氧化氢酶在体内持续产生氧气可以逆转肿瘤缺氧，通过阻断原卟啉 IX 外流（下调缺氧诱导因子-1α和亚铁螯合酶）来增强原卟啉 IX 积累，并上调原卟啉 IX 生物合成（提供外源性 5-氨基乙酰丙酸并上调 ALA-合成酶）。体内荧光/光声双重成像可以同时监测瘤内氧饱和度和原卟啉 IX 代谢动力学。因此，该方法有利于优化不同癌症的治疗参数，以实现Ca ²⁺ /Cu ²⁺干扰增强的可重复光动力治疗，使该治疗诊断贴片在临床实践中具有广阔的前景。