Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2
Mesenchymal stem cells (MSCs) are being exploited in regenerative medicine due to their tri-lineage differentiation and immunomodulation activity. Currently, there are two major challenges when directing the differentiation of MSCs for therapeutic applications. First, chemical and growth factor stra...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Published: |
Wiley Open Access
2017
|
| Subjects: | |
| Online Access: | https://eprints.nottingham.ac.uk/47837/ |
| _version_ | 1848797642225614848 |
|---|---|
| author | Thiagarajan, Lalitha Abu-Awwad, Hosam Al-Deen M. Dixon, James E. |
| author_facet | Thiagarajan, Lalitha Abu-Awwad, Hosam Al-Deen M. Dixon, James E. |
| author_sort | Thiagarajan, Lalitha |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Mesenchymal stem cells (MSCs) are being exploited in regenerative medicine due to their tri-lineage differentiation and immunomodulation activity. Currently, there are two major challenges when directing the differentiation of MSCs for therapeutic applications. First, chemical and growth factor strategies to direct osteogenesis in vivo lack specificity for targeted delivery with desired effects. Second, MSC differentiation by gene therapy is difficult as transfection with existing approaches is clinically impractical (viral transfection) or have low efficacy (lipid-mediated transfection). These challenges can be avoided by directly delivering nonvirally derived recombinant protein transcription factors with the glycosaminoglycan-binding enhanced transduction (GET) delivery system (P21 and 8R peptides). We used the osteogenic master regulator, RUNX2 as a programming factor due to its stage-specific role in osteochondral differentiation pathways. Herein, we engineered GET-fusion proteins and compared sequential osteogenic changes in MSCs, induced by exposure to GET fusion proteins or conventional stimulation methods (dexamethasone and Bone morphogenetic protein 2). By assessing loss of stem cell-surface markers, upregulation of osteogenic genes and matrix mineralization, we demonstrate that GET-RUNX2 efficiently transduces MSCs and triggers osteogenesis by enhancing target gene expression directly. The high transduction efficiency of GET system holds great promise for stem cell therapies by allowing reproducible transcriptional control in stem cells, potentially bypassing problems observed with high-concentration growth-factor or pleiotropic steroid therapies. |
| first_indexed | 2025-11-14T20:07:07Z |
| format | Article |
| id | nottingham-47837 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:07:07Z |
| publishDate | 2017 |
| publisher | Wiley Open Access |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-478372020-05-04T19:19:24Z https://eprints.nottingham.ac.uk/47837/ Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 Thiagarajan, Lalitha Abu-Awwad, Hosam Al-Deen M. Dixon, James E. Mesenchymal stem cells (MSCs) are being exploited in regenerative medicine due to their tri-lineage differentiation and immunomodulation activity. Currently, there are two major challenges when directing the differentiation of MSCs for therapeutic applications. First, chemical and growth factor strategies to direct osteogenesis in vivo lack specificity for targeted delivery with desired effects. Second, MSC differentiation by gene therapy is difficult as transfection with existing approaches is clinically impractical (viral transfection) or have low efficacy (lipid-mediated transfection). These challenges can be avoided by directly delivering nonvirally derived recombinant protein transcription factors with the glycosaminoglycan-binding enhanced transduction (GET) delivery system (P21 and 8R peptides). We used the osteogenic master regulator, RUNX2 as a programming factor due to its stage-specific role in osteochondral differentiation pathways. Herein, we engineered GET-fusion proteins and compared sequential osteogenic changes in MSCs, induced by exposure to GET fusion proteins or conventional stimulation methods (dexamethasone and Bone morphogenetic protein 2). By assessing loss of stem cell-surface markers, upregulation of osteogenic genes and matrix mineralization, we demonstrate that GET-RUNX2 efficiently transduces MSCs and triggers osteogenesis by enhancing target gene expression directly. The high transduction efficiency of GET system holds great promise for stem cell therapies by allowing reproducible transcriptional control in stem cells, potentially bypassing problems observed with high-concentration growth-factor or pleiotropic steroid therapies. Wiley Open Access 2017-11-26 Article PeerReviewed Thiagarajan, Lalitha, Abu-Awwad, Hosam Al-Deen M. and Dixon, James E. (2017) Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2. Stem Cells Translational Medicine, 6 (12). pp. 2146-2159. ISSN 2157-6580 Intracellular transduction; Glycosaminoglycan-binding enhanced transduction; Cell-penetrating peptide; Osteogenesis; RUNX2 https://doi.org/10.1002/sctm.17-0137 doi:10.1002/sctm.17-0137 doi:10.1002/sctm.17-0137 |
| spellingShingle | Intracellular transduction; Glycosaminoglycan-binding enhanced transduction; Cell-penetrating peptide; Osteogenesis; RUNX2 Thiagarajan, Lalitha Abu-Awwad, Hosam Al-Deen M. Dixon, James E. Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title | Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title_full | Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title_fullStr | Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title_full_unstemmed | Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title_short | Osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of RUNX2 |
| title_sort | osteogenic programming of human mesenchymal stem cells with highly efficient intracellular delivery of runx2 |
| topic | Intracellular transduction; Glycosaminoglycan-binding enhanced transduction; Cell-penetrating peptide; Osteogenesis; RUNX2 |
| url | https://eprints.nottingham.ac.uk/47837/ https://eprints.nottingham.ac.uk/47837/ https://eprints.nottingham.ac.uk/47837/ |