Frontiers Science Center for Synthetic Biology, Key Laboratory of Systems Bioengineering (MOE), School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300350, P. R. China.
State Key Laboratory of Systems Medicine for Cancer, Shanghai Cancer Institute, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, 200032, P. R. China.
Adv Mater. 2024 Apr;36(15):e2309534. doi: 10.1002/adma.202309534. Epub 2024 Jan 15.
Photodynamic therapy (PDT) depends on the light-irradiated exciting of photosensitizer (PS) to generate reactive oxygen species (ROS), which faces challenges and limitations in hypoxia and antioxidant response of cancer cells, and limited tissue-penetration of light. Herein, a multifunctional DNA/upconversion nanoparticles (UCNPs) complex is developed which enables controlled co-delivery of CRISPR-Cas9, hemin, and protoporphyrin (PP) for synergistic PDT. An ultralong single-stranded DNA (ssDNA) is prepared via rolling circle amplification (RCA), which contains recognition sequences of single guide RNA (sgRNA) for loading Cas9 ribonucleoprotein (RNP), G-quadruplex sequences for loading hemin and PP, and linker sequences for combining UCNP. Cas9 RNP cleaves the antioxidant regulator nuclear factor E2-related factor 2 (Nrf2), improving the sensitivity of cancer cells to ROS, and enhancing the synergistic PDT effect. The G-quadruplex/hemin DNAzyme mimicks horseradish peroxidase (HRP) to catalyze the endogenous HO to O, overcoming hypoxia condition in tumors. The introduced UCNP converts NIR irradiation with deep tissue penetration to light with shorter wavelength, exciting PP to transform the abundant O to O. The integration of gene editing and PDT allows substantial accumulation of O in cancer cells for enhanced cell apoptosis, and this synergistic PDT has shown remarkable therapeutic efficacy in a breast cancer mouse model.
光动力疗法（PDT）依赖于光敏剂（PS）的光激发来产生活性氧（ROS），但在癌细胞的缺氧和抗氧化反应以及光的组织穿透性有限的情况下，面临着挑战和限制。在此，开发了一种多功能 DNA/上转换纳米粒子（UCNP）复合物，可实现 CRISPR-Cas9、血红素和原卟啉（PP）的控制共递，以实现协同 PDT。通过滚环扩增（RCA）制备了超长长单链 DNA（ssDNA），其中包含用于加载 Cas9 核糖核蛋白（RNP）的单指导 RNA（sgRNA）的识别序列、用于加载血红素和 PP 的 G-四链体序列以及用于结合 UCNP 的连接序列。Cas9 RNP 切割抗氧化调节剂核因子 E2 相关因子 2（Nrf2），提高了癌细胞对 ROS 的敏感性，并增强了协同 PDT 效应。G-四链体/血红素 DNA 酶模拟辣根过氧化物酶（HRP）催化内源性 HO 转化为 O，克服了肿瘤中的缺氧条件。引入的 UCNP 将具有深组织穿透性的近红外辐射转换为具有更短波长的光，激发 PP 将丰富的 O 转化为 O。基因编辑和 PDT 的结合允许 O 在癌细胞中大量积累，以增强细胞凋亡，这种协同 PDT 在乳腺癌小鼠模型中显示出显著的治疗效果。
