Intratracheally administered LNA gapmer antisense oligonucleotides induce robust gene silencing in mouse lung fibroblasts
© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research..
The lung is a complex organ with various cell types having distinct roles. Antisense oligonucleotides (ASOs) have been studied in the lung, but it has been challenging to determine their effectiveness in each cell type due to the lack of appropriate analytical methods. We employed three distinct approaches to study silencing efficacy within different cell types. First, we used lineage markers to identify cell types in flow cytometry, and simultaneously measured ASO-induced silencing of cell-surface proteins CD47 or CD98. Second, we applied single-cell RNA sequencing (scRNA-seq) to measure silencing efficacy in distinct cell types; to the best of our knowledge, this is the first time scRNA-seq has been applied to measure the efficacy of oligonucleotide therapeutics. In both approaches, fibroblasts were the most susceptible to locally delivered ASOs, with significant silencing also in endothelial cells. Third, we confirmed that the robust silencing in fibroblasts is broadly applicable by silencing two targets expressed mainly in fibroblasts, Mfap4 and Adam33. Across independent approaches, we demonstrate that intratracheally administered LNA gapmer ASOs robustly induce gene silencing in lung fibroblasts. ASO-induced gene silencing in fibroblasts was durable, lasting 4-8 weeks after a single dose. Thus, lung fibroblasts are well aligned with ASOs as therapeutics.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:50 |
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| Enthalten in: |
Nucleic acids research - 50(2022), 15 vom: 26. Aug., Seite 8418-8430 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Shin, Minwook [VerfasserIn] |
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| Links: |
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| Themen: |
Journal Article |
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| Anmerkungen: |
Date Completed 29.08.2022 Date Revised 15.09.2022 published: Print Citation Status MEDLINE |
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| doi: |
10.1093/nar/gkac630 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM344399133 |
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| 100 | 1 | |a Shin, Minwook |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Intratracheally administered LNA gapmer antisense oligonucleotides induce robust gene silencing in mouse lung fibroblasts |
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| 520 | |a © The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. | ||
| 520 | |a The lung is a complex organ with various cell types having distinct roles. Antisense oligonucleotides (ASOs) have been studied in the lung, but it has been challenging to determine their effectiveness in each cell type due to the lack of appropriate analytical methods. We employed three distinct approaches to study silencing efficacy within different cell types. First, we used lineage markers to identify cell types in flow cytometry, and simultaneously measured ASO-induced silencing of cell-surface proteins CD47 or CD98. Second, we applied single-cell RNA sequencing (scRNA-seq) to measure silencing efficacy in distinct cell types; to the best of our knowledge, this is the first time scRNA-seq has been applied to measure the efficacy of oligonucleotide therapeutics. In both approaches, fibroblasts were the most susceptible to locally delivered ASOs, with significant silencing also in endothelial cells. Third, we confirmed that the robust silencing in fibroblasts is broadly applicable by silencing two targets expressed mainly in fibroblasts, Mfap4 and Adam33. Across independent approaches, we demonstrate that intratracheally administered LNA gapmer ASOs robustly induce gene silencing in lung fibroblasts. ASO-induced gene silencing in fibroblasts was durable, lasting 4-8 weeks after a single dose. Thus, lung fibroblasts are well aligned with ASOs as therapeutics | ||
| 650 | 4 | |a Journal Article | |
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| 700 | 1 | |a Chan, Io Long |e verfasserin |4 aut | |
| 700 | 1 | |a Cao, Yuming |e verfasserin |4 aut | |
| 700 | 1 | |a Gruntman, Alisha M |e verfasserin |4 aut | |
| 700 | 1 | |a Lee, Jonathan |e verfasserin |4 aut | |
| 700 | 1 | |a Sousa, Jacquelyn |e verfasserin |4 aut | |
| 700 | 1 | |a Rodríguez, Tomás C |e verfasserin |4 aut | |
| 700 | 1 | |a Echeverria, Dimas |e verfasserin |4 aut | |
| 700 | 1 | |a Devi, Gitali |e verfasserin |4 aut | |
| 700 | 1 | |a Debacker, Alexandre J |e verfasserin |4 aut | |
| 700 | 1 | |a Moazami, Michael P |e verfasserin |4 aut | |
| 700 | 1 | |a Krishnamurthy, Pranathi Meda |e verfasserin |4 aut | |
| 700 | 1 | |a Rembetsy-Brown, Julia M |e verfasserin |4 aut | |
| 700 | 1 | |a Kelly, Karen |e verfasserin |4 aut | |
| 700 | 1 | |a Yukselen, Onur |e verfasserin |4 aut | |
| 700 | 1 | |a Donnard, Elisa |e verfasserin |4 aut | |
| 700 | 1 | |a Parsons, Teagan J |e verfasserin |4 aut | |
| 700 | 1 | |a Khvorova, Anastasia |e verfasserin |4 aut | |
| 700 | 1 | |a Sontheimer, Erik J |e verfasserin |4 aut | |
| 700 | 1 | |a Maehr, René |e verfasserin |4 aut | |
| 700 | 1 | |a Garber, Manuel |e verfasserin |4 aut | |
| 700 | 1 | |a Watts, Jonathan K |e verfasserin |4 aut | |
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