Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis
Objective: MicroRNA-140-3p is the most prevalent form of canonical miR-140 in native chondrocytes. IsomiRs are sequence variants of microRNAs with potentially distinct functionalities. Here we present functional studies of canonical microRNA-140-3p and two of its most prevalent isomiRs, a 5′ isomiR...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-09-01
|
| Series: | Osteoarthritis and Cartilage Open |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665913121000522 |
| _version_ | 1818586899081592832 |
|---|---|
| author | Rua Nader Al-Modawi Jan E. Brinchmann Tommy A. Karlsen |
| author_facet | Rua Nader Al-Modawi Jan E. Brinchmann Tommy A. Karlsen |
| author_sort | Rua Nader Al-Modawi |
| collection | DOAJ |
| description | Objective: MicroRNA-140-3p is the most prevalent form of canonical miR-140 in native chondrocytes. IsomiRs are sequence variants of microRNAs with potentially distinct functionalities. Here we present functional studies of canonical microRNA-140-3p and two of its most prevalent isomiRs, a 5′ isomiR and a 3′ isomiR, in an inflammation-induced model of osteoarthritis (OA). Method: Canonical miR-140-3p, the 5′ isomiR and the 3′ isomiR were overexpressed separately in chondrocytes from three donors and subsequently subjected to an inflammatory milieu mediated by interleukin 1 beta and tumor necrosis factor alpha. RNA sequencing was performed on the cells to investigate the altered transcriptomes, RT-qPCR was performed to validate important observations, and western blot analysis was carried out to further study key inflammatory molecules. Results: The three microRNAs downregulated many of the same genes. However, the 5′ isomiR showed a much greater target spectrum compared to the other two miRNAs, and downregulated cascades of genes downstream of interferon beta, interferon gamma and interleukin 1 beta as well as genes involved in several other inflammatory and antiviral pathways. In addition the 5′ isomiR downregulated practically all HLA class II and class I genes. Conclusion: Introduction of the 5′ isomiR led to downregulation of genes essential for some of the most important inflammation cascades and virtual silencing of genes responsible for antigen presentation. These observations may indicate a very promising therapeutic potential for the 5′ isomiR for OA and several inflammatory conditions, particularly HLA associated immune conditions including many arthritic diseases. |
| first_indexed | 2024-12-16T09:00:17Z |
| format | Article |
| id | doaj.art-9d71648e4f3d437b946812c2370e2215 |
| institution | Directory Open Access Journal |
| issn | 2665-9131 |
| language | English |
| last_indexed | 2024-12-16T09:00:17Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Osteoarthritis and Cartilage Open |
| spelling | doaj.art-9d71648e4f3d437b946812c2370e22152022-12-21T22:37:11ZengElsevierOsteoarthritis and Cartilage Open2665-91312021-09-0133100189Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritisRua Nader Al-Modawi0Jan E. Brinchmann1Tommy A. Karlsen2Department of Immunology, Institute of Clinical Medicine, University of Oslo, Norway; Corresponding author. Department of Immunology, Institute of Clinical Medicine, University of Oslo, Post Box 4950 Nydalen, 0424, Oslo, Norway.Department of Immunology and Transfusion Medicine, Oslo University Hospital, Norway; Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, NorwayDepartment of Immunology and Transfusion Medicine, Oslo University Hospital, NorwayObjective: MicroRNA-140-3p is the most prevalent form of canonical miR-140 in native chondrocytes. IsomiRs are sequence variants of microRNAs with potentially distinct functionalities. Here we present functional studies of canonical microRNA-140-3p and two of its most prevalent isomiRs, a 5′ isomiR and a 3′ isomiR, in an inflammation-induced model of osteoarthritis (OA). Method: Canonical miR-140-3p, the 5′ isomiR and the 3′ isomiR were overexpressed separately in chondrocytes from three donors and subsequently subjected to an inflammatory milieu mediated by interleukin 1 beta and tumor necrosis factor alpha. RNA sequencing was performed on the cells to investigate the altered transcriptomes, RT-qPCR was performed to validate important observations, and western blot analysis was carried out to further study key inflammatory molecules. Results: The three microRNAs downregulated many of the same genes. However, the 5′ isomiR showed a much greater target spectrum compared to the other two miRNAs, and downregulated cascades of genes downstream of interferon beta, interferon gamma and interleukin 1 beta as well as genes involved in several other inflammatory and antiviral pathways. In addition the 5′ isomiR downregulated practically all HLA class II and class I genes. Conclusion: Introduction of the 5′ isomiR led to downregulation of genes essential for some of the most important inflammation cascades and virtual silencing of genes responsible for antigen presentation. These observations may indicate a very promising therapeutic potential for the 5′ isomiR for OA and several inflammatory conditions, particularly HLA associated immune conditions including many arthritic diseases.http://www.sciencedirect.com/science/article/pii/S2665913121000522miR-140-3pIsomiRsOsteoarthritisImmune regulationHLA |
| spellingShingle | Rua Nader Al-Modawi Jan E. Brinchmann Tommy A. Karlsen Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis Osteoarthritis and Cartilage Open miR-140-3p IsomiRs Osteoarthritis Immune regulation HLA |
| title | Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis |
| title_full | Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis |
| title_fullStr | Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis |
| title_full_unstemmed | Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis |
| title_short | Extensive downregulation of immune gene expression by microRNA-140-3p 5′ isomiR in an in vitro model of osteoarthritis |
| title_sort | extensive downregulation of immune gene expression by microrna 140 3p 5 isomir in an in vitro model of osteoarthritis |
| topic | miR-140-3p IsomiRs Osteoarthritis Immune regulation HLA |
| url | http://www.sciencedirect.com/science/article/pii/S2665913121000522 |
| work_keys_str_mv | AT ruanaderalmodawi extensivedownregulationofimmunegeneexpressionbymicrorna1403p5isomirinaninvitromodelofosteoarthritis AT janebrinchmann extensivedownregulationofimmunegeneexpressionbymicrorna1403p5isomirinaninvitromodelofosteoarthritis AT tommyakarlsen extensivedownregulationofimmunegeneexpressionbymicrorna1403p5isomirinaninvitromodelofosteoarthritis |