Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1
Alpha-1 antitrypsin-overexpressing mesenchymal stromal/stem cells (AAT-MSCs) showed improved innate properties with a faster proliferation rate when studied for their protective effects in mouse models of diseases. Here, we investigated the potential mechanism(s) by which AAT gene insertion increase...
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MDPI AG
2024-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/4/2015 |
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| author | Bryan Wolf Prasanth Muralidharan Michael Y. Lee Wei Hua Erica Green Hongjun Wang Charlie Strange |
| author_facet | Bryan Wolf Prasanth Muralidharan Michael Y. Lee Wei Hua Erica Green Hongjun Wang Charlie Strange |
| author_sort | Bryan Wolf |
| collection | DOAJ |
| description | Alpha-1 antitrypsin-overexpressing mesenchymal stromal/stem cells (AAT-MSCs) showed improved innate properties with a faster proliferation rate when studied for their protective effects in mouse models of diseases. Here, we investigated the potential mechanism(s) by which AAT gene insertion increases MSC proliferation. Human bone marrow-derived primary or immortalized MSCs (iMSCs) or AAT-MSCs (iAAT-MSCs) were used in the study. Cell proliferation was measured by cell counting and cell cycle analysis. Possible pathways involved in the pro-proliferation effect of AAT were investigated by measuring mRNA and protein expression of key cell cycle genes. Interval cell counting showed increased proliferation in AAT-MSCs or iAAT-MSCs compared to their corresponding MSC controls. Cell cycle analysis revealed more cells progressing into the S and G2/M phases in iAAT-MSCs, with a notable increase in the cell cycle protein, Cyclin D1. Moreover, treatment with Cyclin D1 inhibitors showed that the increase in proliferation is due to Cyclin D1 and that the AAT protein is upstream and a positive regulator of Cyclin D1. Furthermore, AAT’s effect on Cyclin D1 is independent of the Wnt signaling pathway as there were no differences in the expression of regulatory proteins, including GSK3β and β-Catenin in iMSC and iAAT-MSCs. In summary, our results indicate that AAT gene insertion in an immortalized MSC cell line increases cell proliferation and growth by increasing Cyclin D1 expression and consequently causing cells to progress through the cell cycle at a significantly faster rate. |
| first_indexed | 2024-03-07T22:29:58Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-07T22:29:58Z |
| publishDate | 2024-02-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-b8f15a22d58d4a3381ca10787291d4192024-02-23T15:19:23ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-02-01254201510.3390/ijms25042015Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1Bryan Wolf0Prasanth Muralidharan1Michael Y. Lee2Wei Hua3Erica Green4Hongjun Wang5Charlie Strange6Department of Surgery and Medicine, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Surgery and Medicine, Medical University of South Carolina, Charleston, SC 29425, USAAcademic Magnet High School, North Charleston, SC 29405, USADepartment of Surgery and Medicine, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Surgery and Medicine, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Surgery and Medicine, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Medicine, Medical University of South Carolina, CSB 816, 96 Jonathan Lucas St., Charleston, SC 29425, USAAlpha-1 antitrypsin-overexpressing mesenchymal stromal/stem cells (AAT-MSCs) showed improved innate properties with a faster proliferation rate when studied for their protective effects in mouse models of diseases. Here, we investigated the potential mechanism(s) by which AAT gene insertion increases MSC proliferation. Human bone marrow-derived primary or immortalized MSCs (iMSCs) or AAT-MSCs (iAAT-MSCs) were used in the study. Cell proliferation was measured by cell counting and cell cycle analysis. Possible pathways involved in the pro-proliferation effect of AAT were investigated by measuring mRNA and protein expression of key cell cycle genes. Interval cell counting showed increased proliferation in AAT-MSCs or iAAT-MSCs compared to their corresponding MSC controls. Cell cycle analysis revealed more cells progressing into the S and G2/M phases in iAAT-MSCs, with a notable increase in the cell cycle protein, Cyclin D1. Moreover, treatment with Cyclin D1 inhibitors showed that the increase in proliferation is due to Cyclin D1 and that the AAT protein is upstream and a positive regulator of Cyclin D1. Furthermore, AAT’s effect on Cyclin D1 is independent of the Wnt signaling pathway as there were no differences in the expression of regulatory proteins, including GSK3β and β-Catenin in iMSC and iAAT-MSCs. In summary, our results indicate that AAT gene insertion in an immortalized MSC cell line increases cell proliferation and growth by increasing Cyclin D1 expression and consequently causing cells to progress through the cell cycle at a significantly faster rate.https://www.mdpi.com/1422-0067/25/4/2015mesenchymal stromal/stem cellsalpha-1 antitrypsincell proliferationCyclin D1 |
| spellingShingle | Bryan Wolf Prasanth Muralidharan Michael Y. Lee Wei Hua Erica Green Hongjun Wang Charlie Strange Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 International Journal of Molecular Sciences mesenchymal stromal/stem cells alpha-1 antitrypsin cell proliferation Cyclin D1 |
| title | Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 |
| title_full | Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 |
| title_fullStr | Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 |
| title_full_unstemmed | Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 |
| title_short | Overexpression of Alpha-1 Antitrypsin Increases the Proliferation of Mesenchymal Stem Cells by Upregulation of Cyclin D1 |
| title_sort | overexpression of alpha 1 antitrypsin increases the proliferation of mesenchymal stem cells by upregulation of cyclin d1 |
| topic | mesenchymal stromal/stem cells alpha-1 antitrypsin cell proliferation Cyclin D1 |
| url | https://www.mdpi.com/1422-0067/25/4/2015 |
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