Photochemical internalisation of a macromolecular protein toxin using a cell penetrating peptide-photosensitiser conjugate
Photochemical internalisation of a macromolecular protein toxin using a cell penetrating peptide-photosensitiser conjugate
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DOI：
10.1016/j.jconrel.2011.08.025
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发表时间：
2012-01-30
影响因子：
10.8
通讯作者：
MacRobert, Alexander J.
中科院分区：
文献类型：
--
作者：
Wang, Julie T. -W.;Giuntini, Francesca;MacRobert, Alexander J.
光化学内化（PCI）是一种用于改善大分子药物的细胞递送的位点特异性技术。在这项研究中，含有核心 HIV-1 Tat 48-57 序列的细胞穿透肽与卟啉光敏剂结合，已被证明对 PCI 有效。在此，我们报告了阳离子 Tat 肽与疏水性四苯基卟啉衍生物的水溶性生物缀合物的光物理和光生物学特性的研究。在 HN5 人头颈鳞状细胞癌细胞系中检查了两亲性生物缀合物的细胞摄取和定位。使用荧光检测发现了内/溶酶体囊泡中的有效细胞摄取和定位，并且观察到光诱导的囊泡破裂导致更弥散的细胞内荧光分布。因此，Tat 序列与疏水性卟啉的缀合能够实现两亲性光敏剂的细胞递送，然后该光敏剂可以定位于内膜/溶酶体膜中，这是有效 PCI 治疗所需的。 PCI 功效与蛋白质毒素皂草素联合测试，与单独皂草素或光敏剂治疗相比，细胞活力显着降低。这项研究表明，细胞穿透肽-光敏剂生物共轭策略是一种有前途且通用的方法，可通过光化学内化增强生物活性剂的治疗潜力。 (C) 2011 Elsevier B.V. 保留所有权利。
Photochemical internalisation (PCI) is a site-specific technique for improving cellular delivery of macromolecular drugs. In this study, a cell penetrating peptide, containing the core HIV-1 Tat 48-57 sequence, conjugated with a porphyrin photosensitiser has been shown to be effective for PCI. Herein we report an investigation of the photophysical and photobiological properties of a water soluble bioconjugate of the cationic Tat peptide with a hydrophobic tetraphenylporphyrin derivative. The cellular uptake and localisation of the amphiphilic bioconjugate was examined in the HN5 human head and neck squamous cell carcinoma cell line. Efficient cellular uptake and localisation in endo/lysosomal vesicles was found using fluorescence detection, and light-induced, rupture of the vesicles resulting in a more diffuse intracellular fluorescence distribution was observed. Conjugation of the Tat sequence with a hydrophobic porphyrin thus enables cellular delivery of an amphiphilic photosensitiser which can then localise in endo/lysosomal membranes, as required for effective PCI treatment. PCI efficacy was tested in combination with a protein toxin, saporin, and a significant reduction in cell viability was measured versus saporin or photosensitiser treatment alone. This study demonstrates that the cell penetrating peptide-photosensitiser bioconjugation strategy is a promising and versatile approach for enhancing the therapeutic potential of bioactive agents through photochemical internalisation. (C) 2011 Elsevier B.V. All rights reserved.