Cytosolic Delivery of Macromolecules in Live Human Cells Using the Combined Endosomal Escape Activities of a Small Molecule and Cell Penetrating Peptides
Cytosolic Delivery of Macromolecules in Live Human Cells Using the Combined Endosomal Escape Activities of a Small Molecule and Cell Penetrating Peptides
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DOI：
10.1021/acschembio.9b00585
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发表时间：
2019-12-01
影响因子：
4
通讯作者：
Pellois, Jean-Philippe
中科院分区：
文献类型：
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作者：
Allen, Jason;Najjar, Kristina;Pellois, Jean-Philippe
无效的细胞递送是许多生物应用中的常见问题。开发在各种实验环境中稳健工作的递送试剂仍然是一个挑战。在此，我们报告了如何从原型细胞穿透肽达特衍生的肽可以与小分子UNC7938组合使用，以通过简单的共孵育方案将大分子递送到细胞的胞质溶胶中。我们建立了成功的肽，DNA质粒和单链可变片段抗体的交付。我们还证明，交付工程在难以检测的哺乳动物细胞和条件下，通常抑制细胞穿透肽。在机制上，UNC7938使内体的膜不稳定。这反过来又增强了细胞穿透肽的内体渗漏活性，并促进了最初由哺乳动物细胞通过内吞作用内化的大分子的内体逃逸。这种组合的选择性膜去稳定化代表了用于递送工具的新的化学空间，并且为内体截留问题提供了新的解决方案，该问题通常限制了基于试剂的递送方法的有效性。
Ineffective cellular delivery is a common problem in numerous biological applications. Developing delivery reagents that work robustly in a variety of experimental settings remains a challenge. Herein, we report how peptides derived from the prototypical cell penetrating peptide TAT can be used in combination with a small molecule, UNC7938, to deliver macromolecules into the cytosol of cells by a simple co-incubation protocol. We establish successful delivery of peptides, DNA plasmids, and a single-chain variable fragment antibody. We also demonstrate that delivery works in hard-to-transfect mammalian cells and under conditions typically inhibitory to cell-penetrating peptides. Mechanistically, UNC7938 destabilizes the membrane of endosomes. This, in turn, enhances the endosome-leakage activity of cell-penetrating peptides and facilitates the endosomal escape of macromolecules initially internalized by mammalian cells via endocytosis. This combined selective membrane-destabilization represents a new chemical space for delivery tools and provides a novel solution to the problem of endosomal entrapment that often limits the effectiveness of reagent-based delivery approaches.