Golgi-endosome transport mediated by M6PR facilitates release of antisense oligonucleotides from endosomes
| Autor: | Hong Sun, Stanley T. Crooke, Timothy A. Vickers, Cheryl L De Hoyos, Chih-Wei Hsu, Joshua G Nichols, Xue-hai Liang |
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| Rok vydání: | 2019 |
| Předmět: |
Endosome
Golgi Apparatus Phosphorothioate Oligonucleotides Endosomes Biology Receptor IGF Type 2 Mice symbols.namesake Genetics Animals Humans Receptor Molecular Biology Oligonucleotide Golgi Matrix Proteins Oligonucleotides Antisense Golgi apparatus Endocytosis Cell biology Protein Transport Membrane Drug delivery Antisense oligonucleotides symbols Intracellular HeLa Cells |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gkz1171 |
| Popis: | Release of phosphorothioate antisense oligonucleotides (PS-ASOs) from late endosomes (LEs) is a rate-limiting step and a poorly defined process for productive intracellular ASO drug delivery. Here, we examined the role of Golgi-endosome transport, specifically M6PR shuttling mediated by GCC2, in PS-ASO trafficking and activity. We found that reduction in cellular levels of GCC2 or M6PR impaired PS-ASO release from endosomes and decreased PS-ASO activity in human cells. GCC2 relocated to LEs upon PS-ASO treatment, and M6PR also co-localized with PS-ASOs in LEs or on LE membranes. These proteins act through the same pathway to influence PS-ASO activity, with GCC2 action preceding that of M6PR. Our data indicate that M6PR binds PS-ASOs and facilitates their vesicular escape. The co-localization of M6PR and of GCC2 with ASOs is influenced by the PS modifications, which have been shown to enhance the affinity of ASOs for proteins, suggesting that localization of these proteins to LEs is mediated by ASO-protein interactions. Reduction of M6PR levels also decreased PS-ASO activity in mouse cells and in livers of mice treated subcutaneously with PS-ASO, indicating a conserved mechanism. Together, these results demonstrate that the transport machinery between LE and Golgi facilitates PS-ASO release. |
| Databáze: | OpenAIRE |
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