Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability
AimsExosomes (exo) are involved in the progression of osteoarthritis (OA). This study aimed to investigate the function of dysfunctional chondrocyte-derived exo (DC-exo) on OA in rats and rat macrophages.MethodsRat-derived chondrocytes were isolated, and DCs induced with interleukin (IL)-1β were use...
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| Format: | Article |
| Language: | English |
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The British Editorial Society of Bone & Joint Surgery
2022-09-01
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| Series: | Bone & Joint Research |
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| Online Access: | https://online.boneandjoint.org.uk/doi/10.1302/2046-3758.119.BJR-2021-0443.R2 |
| _version_ | 1798001348870930432 |
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| author | Guohua Lv Bing Wang Lei Li Yunchao Li Xinyi Li Haoyu He Lei Kuang |
| author_facet | Guohua Lv Bing Wang Lei Li Yunchao Li Xinyi Li Haoyu He Lei Kuang |
| author_sort | Guohua Lv |
| collection | DOAJ |
| description | AimsExosomes (exo) are involved in the progression of osteoarthritis (OA). This study aimed to investigate the function of dysfunctional chondrocyte-derived exo (DC-exo) on OA in rats and rat macrophages.MethodsRat-derived chondrocytes were isolated, and DCs induced with interleukin (IL)-1β were used for exo isolation. Rats with OA (n = 36) or macrophages were treated with DC-exo or phosphate-buffered saline (PBS). Macrophage polarization and autophagy, and degradation and chondrocyte activity of cartilage tissues, were examined. RNA sequencing was used to detect genes differentially expressed in DC-exo, followed by RNA pull-down and ribonucleoprotein immunoprecipitation (RIP). Long non-coding RNA osteoarthritis non-coding transcript (OANCT) and phosphoinositide-3-kinase regulatory subunit 5 (PIK3R5) were depleted in DC-exo-treated macrophages and OA rats, in order to observe macrophage polarization and cartilage degradation. The PI3K/AKT/mammalian target of rapamycin (mTOR) pathway activity in cells and tissues was measured using western blot.ResultsDC-exo inhibited macrophage autophagy (p = 0.002) and promoted M1 macrophage polarization (p = 0.002). DC-exo at 20 μg/ml induced collagen degradation (p < 0.001) and inflammatory cell infiltration (p = 0.023) in rats. OANCT was elevated in DC (p < 0.001) and in cartilage tissues of OA patients (p < 0.001), and positively correlated with patients’ Kellgren-Lawrence grade (p < 0.001). PIK3R5 was increased in DC-exo-treated cartilage tissues (p < 0.001), and OANCT bound to fat mass and obesity-associated protein (FTO) (p < 0.001). FTO bound to PIK3R5 (p < 0.001) to inhibit the stability of PIK3R5 messenger RNA (mRNA) (p < 0.001) and disrupt the PI3K/AKT/mTOR pathway (p < 0.001).ConclusionExosomal OANCT from DC could bind to FTO protein, thereby maintaining the mRNA stability of PIK3R5, further activating the PI3K/AKT/mTOR pathway to exacerbate OA.Cite this article: Bone Joint Res 2022;11(9):652–668. |
| first_indexed | 2024-04-11T11:34:38Z |
| format | Article |
| id | doaj.art-70faeec33e7e455f8378b3c80275f171 |
| institution | Directory Open Access Journal |
| issn | 2046-3758 |
| language | English |
| last_indexed | 2024-04-11T11:34:38Z |
| publishDate | 2022-09-01 |
| publisher | The British Editorial Society of Bone & Joint Surgery |
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| series | Bone & Joint Research |
| spelling | doaj.art-70faeec33e7e455f8378b3c80275f1712022-12-22T04:26:01ZengThe British Editorial Society of Bone & Joint SurgeryBone & Joint Research2046-37582022-09-0111965266810.1302/2046-3758.119.BJR-2021-0443.R2Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stabilityGuohua Lv0Bing Wang1Lei Li2Yunchao Li3Xinyi Li4Haoyu He5Lei Kuang6Department of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaDepartment of Spine Surgery, The Second Xiangya Hospital of Central South University, Changsha, ChinaAimsExosomes (exo) are involved in the progression of osteoarthritis (OA). This study aimed to investigate the function of dysfunctional chondrocyte-derived exo (DC-exo) on OA in rats and rat macrophages.MethodsRat-derived chondrocytes were isolated, and DCs induced with interleukin (IL)-1β were used for exo isolation. Rats with OA (n = 36) or macrophages were treated with DC-exo or phosphate-buffered saline (PBS). Macrophage polarization and autophagy, and degradation and chondrocyte activity of cartilage tissues, were examined. RNA sequencing was used to detect genes differentially expressed in DC-exo, followed by RNA pull-down and ribonucleoprotein immunoprecipitation (RIP). Long non-coding RNA osteoarthritis non-coding transcript (OANCT) and phosphoinositide-3-kinase regulatory subunit 5 (PIK3R5) were depleted in DC-exo-treated macrophages and OA rats, in order to observe macrophage polarization and cartilage degradation. The PI3K/AKT/mammalian target of rapamycin (mTOR) pathway activity in cells and tissues was measured using western blot.ResultsDC-exo inhibited macrophage autophagy (p = 0.002) and promoted M1 macrophage polarization (p = 0.002). DC-exo at 20 μg/ml induced collagen degradation (p < 0.001) and inflammatory cell infiltration (p = 0.023) in rats. OANCT was elevated in DC (p < 0.001) and in cartilage tissues of OA patients (p < 0.001), and positively correlated with patients’ Kellgren-Lawrence grade (p < 0.001). PIK3R5 was increased in DC-exo-treated cartilage tissues (p < 0.001), and OANCT bound to fat mass and obesity-associated protein (FTO) (p < 0.001). FTO bound to PIK3R5 (p < 0.001) to inhibit the stability of PIK3R5 messenger RNA (mRNA) (p < 0.001) and disrupt the PI3K/AKT/mTOR pathway (p < 0.001).ConclusionExosomal OANCT from DC could bind to FTO protein, thereby maintaining the mRNA stability of PIK3R5, further activating the PI3K/AKT/mTOR pathway to exacerbate OA.Cite this article: Bone Joint Res 2022;11(9):652–668.https://online.boneandjoint.org.uk/doi/10.1302/2046-3758.119.BJR-2021-0443.R2OsteoarthritisDysfunctional chondrocyte-derived exosomesLong non-coding RNA OANCTPIK3R5FTOOsteoarthritis (OA) |
| spellingShingle | Guohua Lv Bing Wang Lei Li Yunchao Li Xinyi Li Haoyu He Lei Kuang Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability Bone & Joint Research Osteoarthritis Dysfunctional chondrocyte-derived exosomes Long non-coding RNA OANCT PIK3R5 FTO Osteoarthritis (OA) |
| title | Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability |
| title_full | Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability |
| title_fullStr | Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability |
| title_full_unstemmed | Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability |
| title_short | Exosomes from dysfunctional chondrocytes affect osteoarthritis in Sprague-Dawley rats through FTO-dependent regulation of PIK3R5 mRNA stability |
| title_sort | exosomes from dysfunctional chondrocytes affect osteoarthritis in sprague dawley rats through fto dependent regulation of pik3r5 mrna stability |
| topic | Osteoarthritis Dysfunctional chondrocyte-derived exosomes Long non-coding RNA OANCT PIK3R5 FTO Osteoarthritis (OA) |
| url | https://online.boneandjoint.org.uk/doi/10.1302/2046-3758.119.BJR-2021-0443.R2 |
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