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A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity
Endocytosis is a mechanism by which cells sense their environment and internalize various nutrients, growth factors and signaling molecules. This process initiates at the plasma membrane, converges with autophagy, and terminates at the lysosome. It is well-established that cellular uptake of antisen...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6868497/ https://www.ncbi.nlm.nih.gov/pubmed/31612951 http://dx.doi.org/10.1093/nar/gkz901 |
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author | Ochaba, Joseph Powers, Andrew F Tremble, Kaitlyn A Greenlee, Sarah Post, Noah M Matson, John E MacLeod, A Robert Guo, Shuling Aghajan, Mariam |
author_facet | Ochaba, Joseph Powers, Andrew F Tremble, Kaitlyn A Greenlee, Sarah Post, Noah M Matson, John E MacLeod, A Robert Guo, Shuling Aghajan, Mariam |
author_sort | Ochaba, Joseph |
collection | PubMed |
description | Endocytosis is a mechanism by which cells sense their environment and internalize various nutrients, growth factors and signaling molecules. This process initiates at the plasma membrane, converges with autophagy, and terminates at the lysosome. It is well-established that cellular uptake of antisense oligonucleotides (ASOs) proceeds through the endocytic pathway; however, only a small fraction escapes endosomal trafficking while the majority are rendered inactive in the lysosome. Since these pathways converge and share common molecular machinery, it is unclear if autophagy-related trafficking participates in ASO uptake or whether modulation of autophagy affects ASO activity and localization. To address these questions, we investigated the effects of autophagy modulation on ASO activity in cells and mice. We found that enhancing autophagy through small-molecule mTOR inhibition, serum-starvation/fasting, and ketogenic diet, increased ASO-mediated target reduction in vitro and in vivo. Additionally, autophagy activation enhanced the localization of ASOs into autophagosomes without altering intracellular concentrations or trafficking to other compartments. These results support a novel role for autophagy and the autophagosome as a previously unidentified compartment that participates in and contributes to enhanced ASO activity. Further, we demonstrate non-chemical methods to enhance autophagy and subsequent ASO activity using translatable approaches such as fasting or ketogenic diet. |
format | Online Article Text |
id | pubmed-6868497 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-68684972019-11-27 A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity Ochaba, Joseph Powers, Andrew F Tremble, Kaitlyn A Greenlee, Sarah Post, Noah M Matson, John E MacLeod, A Robert Guo, Shuling Aghajan, Mariam Nucleic Acids Res Molecular Biology Endocytosis is a mechanism by which cells sense their environment and internalize various nutrients, growth factors and signaling molecules. This process initiates at the plasma membrane, converges with autophagy, and terminates at the lysosome. It is well-established that cellular uptake of antisense oligonucleotides (ASOs) proceeds through the endocytic pathway; however, only a small fraction escapes endosomal trafficking while the majority are rendered inactive in the lysosome. Since these pathways converge and share common molecular machinery, it is unclear if autophagy-related trafficking participates in ASO uptake or whether modulation of autophagy affects ASO activity and localization. To address these questions, we investigated the effects of autophagy modulation on ASO activity in cells and mice. We found that enhancing autophagy through small-molecule mTOR inhibition, serum-starvation/fasting, and ketogenic diet, increased ASO-mediated target reduction in vitro and in vivo. Additionally, autophagy activation enhanced the localization of ASOs into autophagosomes without altering intracellular concentrations or trafficking to other compartments. These results support a novel role for autophagy and the autophagosome as a previously unidentified compartment that participates in and contributes to enhanced ASO activity. Further, we demonstrate non-chemical methods to enhance autophagy and subsequent ASO activity using translatable approaches such as fasting or ketogenic diet. Oxford University Press 2019-12-02 2019-10-15 /pmc/articles/PMC6868497/ /pubmed/31612951 http://dx.doi.org/10.1093/nar/gkz901 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Molecular Biology Ochaba, Joseph Powers, Andrew F Tremble, Kaitlyn A Greenlee, Sarah Post, Noah M Matson, John E MacLeod, A Robert Guo, Shuling Aghajan, Mariam A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title | A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title_full | A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title_fullStr | A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title_full_unstemmed | A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title_short | A novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
title_sort | novel and translational role for autophagy in antisense oligonucleotide trafficking and activity |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6868497/ https://www.ncbi.nlm.nih.gov/pubmed/31612951 http://dx.doi.org/10.1093/nar/gkz901 |
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