Endocytosis‐Independent and Cancer‐Selective Cytosolic Protein Delivery via Reversible Tagging with LAT1 substrate. Issue 35 (28th July 2022)

• Record Type: Journal Article
• Authors: Zhao, Ziyin; Liu, Xun; Hou, Mengying; Zhou, Renxiang; Wu, Fan; Yan, Jing; Li, Wei; Zheng, Yujia; Zhong, Qinmeng; Chen, Yongbing; Yin, Lichen
• Is Part Of: Advanced materials. Volume 34:Issue 35(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-07-28
• Subjects: carrier‐free delivery -- cytosolic protein delivery -- endocytosis‐free internalization -- l‐type amino acid transporter 1 (LAT1) -- pro‐protein -- reactive oxygen species (ROS)‐responsiveness; Materials -- Periodicals; Chemical vapor deposition -- Periodicals; 620.11
• DOI: 10.1002/adma.202110560
• Languages: English
• ISSNs: 0935-9648

Abstract

Abstract: Protein drugs targeting intracellular machineries have shown profound therapeutic potentials, but their clinical utilities are greatly hampered by the lack of efficient cytosolic delivery techniques. Existing strategies mainly rely on nanocarriers or conjugated cell‐penetrating peptides (CPPs), which often have drawbacks such as materials complexity/toxicity, lack of cell specificity, and endolysosomal entrapment. Herein, a unique carrier‐free approach is reported for mediating cancer‐selective and endocytosis‐free cytosolic protein delivery. Proteins are sequentially modified with 4‐nitrophenyl 4‐(4, 4, 5, 5‐tetramethyl‐1, 3, 2‐dioxaborolan‐2‐yl) benzyl carbonate as the H2 O2 ‐responsive domain and 3, 4‐dihydroxy‐l ‐phenylalanine as the substrate of l‐ type amino acid transporter 1 (LAT1). Thus, the pro‐protein can be directly transported into tumor cells by overexpressed LAT1 on cell membranes, bypassing endocytosis and endolysosomal entrapment. In the cytosol, overproduced H2 O2 restores the protein structure and activity. Using this technique, versatile proteins are delivered into tumor cells with robust efficiency, including toxins, enzymes, CRISPR‐Cas9 ribonucleoprotein, and antibodies. Furthermore, intravenously injected pro‐protein of saporin shows potent anticancer efficacy in 4T1‐tumor‐bearing mice, without provoking systemic toxicity. Such a facile and versatile pro‐protein platform may benefit the development of protein pharmaceuticals. Abstract : A pro‐protein approach is developed based on reversible tagging with l ‐type amino acid transporter 1 (LAT1) substrate, which enables carrier‐free, cancer‐selective, and endocytosis‐independent cytosolic delivery via LAT1‐mediated direct transport. It demonstrates high generality and robust efficiency for varieties of functional proteins, including enzymes, toxins, antibodies, and CRISPR‐Cas9 ribonucleoprotein. This facile and versatile platform holds transformative potentials in protein pharmaceuticals.