GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells
Targeted delivery of antisense oligonucleotide (AON) drugs is a promising strategy to increase their concentration in the desired tissues and cell types while reducing access to other organs. Conjugation of AONs to N-acetylgalactosamine (GalNAc) has been shown to efficiently shift their biodistribut...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2019-03-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253118302993 |
| _version_ | 1819207706550992896 |
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| author | Sabine Sewing Marcel Gubler Régine Gérard Blandine Avignon Yasmin Mueller Annamaria Braendli-Baiocco Marielle Odin Annie Moisan |
| author_facet | Sabine Sewing Marcel Gubler Régine Gérard Blandine Avignon Yasmin Mueller Annamaria Braendli-Baiocco Marielle Odin Annie Moisan |
| author_sort | Sabine Sewing |
| collection | DOAJ |
| description | Targeted delivery of antisense oligonucleotide (AON) drugs is a promising strategy to increase their concentration in the desired tissues and cell types while reducing access to other organs. Conjugation of AONs to N-acetylgalactosamine (GalNAc) has been shown to efficiently shift their biodistribution toward the liver via high-affinity binding to the asialoglycoprotein receptor (ASGPR) expressed at the surface of hepatocytes. Nevertheless, GalNAc conjugation does not prevent accumulation of AONs in the kidney cortex, and GalNAc-conjugated AONs might cause kidney toxicities, for example, under conditions of ASGPR saturation. Here, we investigated the nephrotoxicity potential of GalNAc-conjugated AONs by in vitro profiling of AON libraries in renal proximal tubule epithelial cells (PTECs) and in vivo testing of selected candidates. Whereas GalNAc-conjugated AONs appeared generally innocuous to PTECs, some caused mild-to-moderate nephrotoxicity in rats. Interestingly, the in vivo kidney liabilities could be recapitulated in vitro by treating PTECs with the unconjugated (or naked) parental AONs. An in vitro mechanistic study revealed that GalNAc conjugation attenuated AON-induced renal cell toxicity despite intracellular accumulation similar to that of naked AONs and independent of target knockdown. Overall, our in vitro findings reveal ASGPR-independent properties of GalNAc AONs that confer a favorable safety profile at the cellular level, which may variably translate in vivo due to catabolic transformation of circulating AONs. Keywords: antisense oligonucleotide, targeted delivery, kidney, ASGPR, toxicity |
| first_indexed | 2024-12-23T05:27:45Z |
| format | Article |
| id | doaj.art-2372d539f88947098a3a24ddd5408f46 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-23T05:27:45Z |
| publishDate | 2019-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-2372d539f88947098a3a24ddd5408f462022-12-21T17:58:33ZengElsevierMolecular Therapy: Nucleic Acids2162-25312019-03-01146779GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular CellsSabine Sewing0Marcel Gubler1Régine Gérard2Blandine Avignon3Yasmin Mueller4Annamaria Braendli-Baiocco5Marielle Odin6Annie Moisan7Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, SwitzerlandRoche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland; Corresponding author: Annie Moisan, PhD, Roche Pharma Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, Basel 4070, Switzerland.Targeted delivery of antisense oligonucleotide (AON) drugs is a promising strategy to increase their concentration in the desired tissues and cell types while reducing access to other organs. Conjugation of AONs to N-acetylgalactosamine (GalNAc) has been shown to efficiently shift their biodistribution toward the liver via high-affinity binding to the asialoglycoprotein receptor (ASGPR) expressed at the surface of hepatocytes. Nevertheless, GalNAc conjugation does not prevent accumulation of AONs in the kidney cortex, and GalNAc-conjugated AONs might cause kidney toxicities, for example, under conditions of ASGPR saturation. Here, we investigated the nephrotoxicity potential of GalNAc-conjugated AONs by in vitro profiling of AON libraries in renal proximal tubule epithelial cells (PTECs) and in vivo testing of selected candidates. Whereas GalNAc-conjugated AONs appeared generally innocuous to PTECs, some caused mild-to-moderate nephrotoxicity in rats. Interestingly, the in vivo kidney liabilities could be recapitulated in vitro by treating PTECs with the unconjugated (or naked) parental AONs. An in vitro mechanistic study revealed that GalNAc conjugation attenuated AON-induced renal cell toxicity despite intracellular accumulation similar to that of naked AONs and independent of target knockdown. Overall, our in vitro findings reveal ASGPR-independent properties of GalNAc AONs that confer a favorable safety profile at the cellular level, which may variably translate in vivo due to catabolic transformation of circulating AONs. Keywords: antisense oligonucleotide, targeted delivery, kidney, ASGPR, toxicityhttp://www.sciencedirect.com/science/article/pii/S2162253118302993 |
| spellingShingle | Sabine Sewing Marcel Gubler Régine Gérard Blandine Avignon Yasmin Mueller Annamaria Braendli-Baiocco Marielle Odin Annie Moisan GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells Molecular Therapy: Nucleic Acids |
| title | GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells |
| title_full | GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells |
| title_fullStr | GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells |
| title_full_unstemmed | GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells |
| title_short | GalNAc Conjugation Attenuates the Cytotoxicity of Antisense Oligonucleotide Drugs in Renal Tubular Cells |
| title_sort | galnac conjugation attenuates the cytotoxicity of antisense oligonucleotide drugs in renal tubular cells |
| url | http://www.sciencedirect.com/science/article/pii/S2162253118302993 |
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