miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction
Background The poor viability of transplanted mesenchymal stem cells (MSCs) hampers their therapeutic efficacy for ischemic heart disease. MicroRNAs are involved in regulation of MSC survival and function. The present study was designed to investigate the molecular effects of miR‐15a/15b on MSC surv...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2019-01-01
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| Series: | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| Subjects: | |
| Online Access: | https://www.ahajournals.org/doi/10.1161/JAHA.118.010157 |
| _version_ | 1811332320598163456 |
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| author | Yingfeng Tu Yan Qiu Li Liu Tao Huang Hao Tang Youbin Liu Wenguang Guo Hongchi Jiang Yuhua Fan Bo Yu |
| author_facet | Yingfeng Tu Yan Qiu Li Liu Tao Huang Hao Tang Youbin Liu Wenguang Guo Hongchi Jiang Yuhua Fan Bo Yu |
| author_sort | Yingfeng Tu |
| collection | DOAJ |
| description | Background The poor viability of transplanted mesenchymal stem cells (MSCs) hampers their therapeutic efficacy for ischemic heart disease. MicroRNAs are involved in regulation of MSC survival and function. The present study was designed to investigate the molecular effects of miR‐15a/15b on MSC survival, focusing on the role of vascular endothelial growth factor receptor 2. Methods and Results We first harvested donor luc(Luciferase)‐MSCs (5×105) isolated from the luciferase transgenic mice with FVB background. Luc‐MSCs were transfected with miR‐15a/15b mimics or inhibitors and cultured under oxygen glucose deprivation condition for 12 hours to mimics the harsh microenvironment in infarcted heart; they were subjected to MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide?Thiazolyl Blue Tetrazolium Bromide) assay, bioluminescence imaging, quantitative reverse transcription–polymerase chain reaction, transferase‐mediated deoxyuridine triphosphate–digoxigenin nick‐end labeling assay, and flow cytometry. Furthermore, the levels of vascular endothelial growth factor receptor 2, protein kinase B, p(Phosphorylate)‐protein kinase B, Bcl‐2, Bax, and caspase‐3 proteins were available by Western blotting assay. In vivo, acute myocardial infarction was induced in 24 mice by coronary ligation, with subsequent receipt of Luc‐MSCs, Luc‐MSCs+miR‐15a/15b inhibitors, or PBS treatment. The therapeutic procedure and treatment effects were tracked and assessed using bioluminescence imaging and echocardiographic measurement. Next, ex vivo imaging and immunohistochemistry were conducted to verify the distribution of MSCs. We demonstrated that miR‐15a/15b targeted vascular endothelial growth factor receptor 2 to modulate MSC survival, possibly via phosphatidylinositol 3‐kinase/protein kinase B signaling pathway, which was proved by bioluminescence imaging, immunohistochemistry analysis, and echocardiographic measurement. Conclusions Luc‐MSCs could be followed dynamically in vitro and in vivo by bioluminescence imaging, and the role of miR‐15a/b could be inferred from the loss of signals from luc‐MSCs. This finding may have practical clinical implications in miR‐15a/15b–modified MSC transplantation in treating myocardial infarction. |
| first_indexed | 2024-04-13T16:34:14Z |
| format | Article |
| id | doaj.art-8974b22334214cf6a4f3fbe4884b7ce0 |
| institution | Directory Open Access Journal |
| issn | 2047-9980 |
| language | English |
| last_indexed | 2024-04-13T16:34:14Z |
| publishDate | 2019-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease |
| spelling | doaj.art-8974b22334214cf6a4f3fbe4884b7ce02022-12-22T02:39:28ZengWileyJournal of the American Heart Association: Cardiovascular and Cerebrovascular Disease2047-99802019-01-018110.1161/JAHA.118.010157miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial InfarctionYingfeng Tu0Yan Qiu1Li Liu2Tao Huang3Hao Tang4Youbin Liu5Wenguang Guo6Hongchi Jiang7Yuhua Fan8Bo Yu9Department of Cardiology The 2nd Hospital of Harbin Medical University Nangang District Harbin ChinaDepartment of Geriatrics Huadong sanatorium Wuxi City Jiangsu Province ChinaDepartment of Anesthesiology The Third Hospital of Harbin Medical University Harbin Heilongjiang ChinaDepartment of Radiology The Fourth Hospital of Harbin Medical University Harbin ChinaDepartment of Cardiology The 2nd Hospital of Harbin Medical University Nangang District Harbin ChinaDepartment of Cardiology The 2nd Hospital of Harbin Medical University Nangang District Harbin ChinaCollege of Basic Medical Science Harbin Medical University–Daqing Daqing ChinaKey Laboratory of Hepatosplenic Surgery Department of General Surgery The First Affiliated Hospital of Harbin Medical University Harbin ChinaCollege of Pharmacy Harbin Medical University–Daqing Daqing ChinaDepartment of Cardiology The 2nd Hospital of Harbin Medical University Nangang District Harbin ChinaBackground The poor viability of transplanted mesenchymal stem cells (MSCs) hampers their therapeutic efficacy for ischemic heart disease. MicroRNAs are involved in regulation of MSC survival and function. The present study was designed to investigate the molecular effects of miR‐15a/15b on MSC survival, focusing on the role of vascular endothelial growth factor receptor 2. Methods and Results We first harvested donor luc(Luciferase)‐MSCs (5×105) isolated from the luciferase transgenic mice with FVB background. Luc‐MSCs were transfected with miR‐15a/15b mimics or inhibitors and cultured under oxygen glucose deprivation condition for 12 hours to mimics the harsh microenvironment in infarcted heart; they were subjected to MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2‐H‐tetrazolium bromide?Thiazolyl Blue Tetrazolium Bromide) assay, bioluminescence imaging, quantitative reverse transcription–polymerase chain reaction, transferase‐mediated deoxyuridine triphosphate–digoxigenin nick‐end labeling assay, and flow cytometry. Furthermore, the levels of vascular endothelial growth factor receptor 2, protein kinase B, p(Phosphorylate)‐protein kinase B, Bcl‐2, Bax, and caspase‐3 proteins were available by Western blotting assay. In vivo, acute myocardial infarction was induced in 24 mice by coronary ligation, with subsequent receipt of Luc‐MSCs, Luc‐MSCs+miR‐15a/15b inhibitors, or PBS treatment. The therapeutic procedure and treatment effects were tracked and assessed using bioluminescence imaging and echocardiographic measurement. Next, ex vivo imaging and immunohistochemistry were conducted to verify the distribution of MSCs. We demonstrated that miR‐15a/15b targeted vascular endothelial growth factor receptor 2 to modulate MSC survival, possibly via phosphatidylinositol 3‐kinase/protein kinase B signaling pathway, which was proved by bioluminescence imaging, immunohistochemistry analysis, and echocardiographic measurement. Conclusions Luc‐MSCs could be followed dynamically in vitro and in vivo by bioluminescence imaging, and the role of miR‐15a/b could be inferred from the loss of signals from luc‐MSCs. This finding may have practical clinical implications in miR‐15a/15b–modified MSC transplantation in treating myocardial infarction.https://www.ahajournals.org/doi/10.1161/JAHA.118.010157cardiaccardiac contractility and energeticscardiac developmentcardiac dysfunction |
| spellingShingle | Yingfeng Tu Yan Qiu Li Liu Tao Huang Hao Tang Youbin Liu Wenguang Guo Hongchi Jiang Yuhua Fan Bo Yu miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease cardiac cardiac contractility and energetics cardiac development cardiac dysfunction |
| title | miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction |
| title_full | miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction |
| title_fullStr | miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction |
| title_full_unstemmed | miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction |
| title_short | miR‐15a/15b Cluster Modulates Survival of Mesenchymal Stem Cells to Improve Its Therapeutic Efficacy of Myocardial Infarction |
| title_sort | mir 15a 15b cluster modulates survival of mesenchymal stem cells to improve its therapeutic efficacy of myocardial infarction |
| topic | cardiac cardiac contractility and energetics cardiac development cardiac dysfunction |
| url | https://www.ahajournals.org/doi/10.1161/JAHA.118.010157 |
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