MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus
Daoyuan Hu1,2, Yunlong Ge1, Yuhang Xi1
1Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Purpose:
The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated.
Materials And Methods:
The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system.
Results:
In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2'-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253.
Conclusions:
Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.
Insights
MicroRNA-145 (miR-145) gene modification enhances bone marrow-derived mesenchymal stem cell (BMSC) retention in corpus cavernosum for erectile dysfunction treatment. This approach modulates the miR-145/KLF4 axis, improving stem cell therapy efficacy.
Area of Science:
- Regenerative Medicine
- Molecular Biology
- Urology
Background:
- Stem cell (SC) retention and differentiation in the corpus cavernosum (CC) are critical for treating erectile dysfunction (ED).
- Poor SC engraftment limits the efficacy of cell-based therapies for ED.
Purpose of the Study:
- To investigate the effects and mechanism of microRNA-145 (miR-145) gene modification on bone marrow-derived mesenchymal stem cells (BMSCs).
- To assess the impact of miR-145 on BMSC traits, fate, and therapeutic potential in an ED model.
Main Methods:
- In vitro analysis of miR-145 effects on BMSC apoptosis, proliferation, migration, and differentiation.
- In vivo transplantation of miR-145 modified BMSCs into age-related ED rat models.
- Verification of miR-145 target gene KLF4 using qPCR and immunohistochemistry.
- Rescue experiments using APTO-253 in co-cultured corpus cavernosum smooth muscle cells (CCSMCs).
Main Results:
- miR-145 inhibited BMSC apoptosis and migration while promoting smooth muscle-like differentiation in vitro.
- In vivo, miR-145 modified BMSCs showed enhanced and sustained retention within the CC.
- miR-145 dampened KLF4 expression in CCSMCs, an effect reversible by APTO-253.
Conclusions:
- miR-145 modification prolongs transplanted BMSC retention in the CC, potentially via the miR-145/KLF4 axis.
- This strategy offers a promising approach to improve stem cell-based treatments for ED.
- Targeted gene modification of stem cells presents an innovative avenue for regenerative medicine in urology.
Related Concept Videos
01:19Mesenchymal Stem Cells
01:22MicroRNAs