Intra- and trans-cellular delivery of enzymes by direct conjugation with non-multivalent anti-ICAM molecules

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• 作者：Rasa Ghaffarian^a ; Niksa Roki^a ; Abraham Abouzeid^c ; Wyatt Vreeland^c ; Silvia Muro^a ; ^b ; ^{muro@umd.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Drug targeting ; monomeric vs. multivalent targeting ; Receptor-mediated endocytosis ; Transcytosis ; Antibody-enzyme conjugate ; ICAM-1
• 刊名：Journal of Controlled Release
• 出版年：2016
• 出版时间：28 September 2016
• 年：2016
• 卷：238
• 期：Complete
• 页码：221-230
• 全文大小：1595 K

文摘

Intercellular adhesion molecule 1 (ICAM-1) is a cell-surface protein overexpressed in many diseases and explored for endocytosis and transcytosis of drug delivery systems. All previous evidence demonstrating ICAM-1-mediated transport of therapeutics into or across cells was obtained using nanocarriers or conjugates coupled to multiple copies of anti-ICAM antibodies or peptides. Yet, transport of therapeutics linked to non-multivalent anti-ICAM ligands has never been shown, since multivalency was believed to be necessary to induce transport. Our goal was to explore whether non-multivalent binding to ICAM-1 could drive endocytosis and/or transcytosis of model cargo in different cell types. We found that anti-ICAM was specifically internalized by all tested ICAM-1-expressing cells, including epithelial, fibroblast and neuroblastoma cells, primary or established cell lines. Uptake was inhibited at 4 °C and in the presence of an inhibitor of the ICAM-1-associated pathway, rather than inhibitors of the clathrin or caveolar routes. We observed minimal transport of anti-ICAM to lysosomes, yet prominent and specific transcytosis across epithelial monolayers. Finally, we coupled a model cargo (the enzyme horseradish peroxidase (HRP)) to anti-ICAM and separated a 1:2 antibody:enzyme conjugate for non-multivalent ICAM-1 targeting. Similar to anti-ICAM, anti-ICAM-HRP was specifically internalized and transported across cells, which rendered intra- and trans-cellular enzyme activity. Therefore, non-multivalent ICAM-1 targeting also provides transport of cargoes into and across cells, representing a new alternative for future therapeutic applications via this route.