Assessing the Effects of Dasatinib on Mesenchymal Stem/Stromal Cells

Medicine Life Sciences Bioengineering Orthopedics Cell Biology
Mesenchymal stem cells Senescence Senolytics Dasatinib Cell therapy

Academic Background

Mesenchymal Stem/Stromal Cells (MSCs) are a type of multipotent stem cells widely used in regenerative medicine and cell therapy. However, MSCs undergo progressive aging (senescence) during in vitro culture, leading to a decline in their proliferation and differentiation capabilities, which poses a significant obstacle to their clinical application. Senescent cells are irreversibly non-dividing cells with significantly reduced functionality, affecting the overall therapeutic efficacy of the cell population. In recent years, researchers have discovered that senolytic drugs (such as Dasatinib) can selectively eliminate senescent cells, thereby preserving and enhancing the functionality of healthy MSCs. Dasatinib is an FDA-approved tyrosine kinase inhibitor primarily used to treat chronic myeloid leukemia, and it has recently been identified as having the potential to clear senescent cells. This study aims to explore the effects of Dasatinib on the phenotypic, genotypic, and immunomodulatory functions of MSCs, providing new strategies for their clinical application.

Source of the Paper

This study was conducted by David P. Heinrichs, Vitali V. Maldonado, I. Kade K. Ardana, Ryan M. Porter, and Rebekah M. Samsonraj. The authors are affiliated with the Department of Biomedical Engineering at the University of Arkansas, the Interdisciplinary Program in Cell and Molecular Biology at the University of Arkansas, and the Department of Orthopedic Surgery at the University of Arkansas for Medical Sciences. The paper was published online on October 29, 2024, in the journal Cellular and Molecular Bioengineering, with the DOI 10.1007/s12195-024-00830-1.

Research Process and Results

1. MSC Culture and Senescence Induction

The study used commercially sourced bone marrow-derived MSCs, with experiments conducted on both young (passages 5–9) and senescent (passages 15–20) cells. Replicative senescence was successfully induced through repeated subculturing. The experiment was divided into four groups: non-senescent cells + Dasatinib, non-senescent cells + DMSO (control), senescent cells + Dasatinib, and senescent cells + DMSO. Each group was performed in triplicate for technical replicates.

2. Live/Dead Staining and Cell Growth Analysis

Through live/dead staining experiments, researchers found that Dasatinib treatment significantly reduced the survival rate of senescent cells, with minimal impact on non-senescent cells. Growth curve analysis showed that cells treated with Dasatinib exhibited significant growth trends over six days, indicating that Dasatinib can clear senescent cells and promote the proliferation of healthy cells.

3. Senescence-Associated β-Galactosidase Staining

Senescence-associated β-galactosidase (SA-β-gal) staining further confirmed that Dasatinib treatment significantly reduced the proportion of senescent cells, while the proportion of non-senescent cells increased significantly. This demonstrates Dasatinib’s ability to selectively eliminate senescent cells.

4. Osteogenic Differentiation and Gene Expression Analysis

In osteogenic differentiation experiments, researchers found that Dasatinib treatment did not significantly affect the osteogenic differentiation capacity of MSCs. Real-time quantitative PCR (RT-qPCR) analysis revealed that Dasatinib treatment significantly upregulated the expression of osteogenic-related genes (e.g., COL1A1 and SP7) in senescent cells, suggesting that Dasatinib may enhance the osteogenic differentiation potential of senescent cells.

5. Adipogenic Differentiation and Gene Expression Analysis

In adipogenic differentiation experiments, Dasatinib treatment did not significantly affect the adipogenic differentiation capacity of MSCs. However, gene expression analysis showed that Dasatinib treatment significantly upregulated the expression of adipogenic-related genes (e.g., PPARγ and CEBPα), indicating that Dasatinib may enhance the adipogenic differentiation potential of MSCs.

6. Chondrogenic Differentiation Experiment

In chondrogenic differentiation experiments, Dasatinib treatment did not significantly affect the chondrogenic differentiation capacity of MSCs. The TGF-β-treated group showed higher glycosaminoglycan (GAG) deposition, but Dasatinib did not significantly alter this phenomenon.

7. Immunomodulatory Function Analysis

Through indoleamine 2,3-dioxygenase (IDO) activity assays, researchers found that Dasatinib treatment did not significantly affect the immunomodulatory function of MSCs. This indicates that Dasatinib treatment does not compromise the immunosuppressive capacity of MSCs.

Research Conclusions and Significance

This study demonstrates that Dasatinib can effectively clear senescent MSCs while preserving and enhancing the proliferation and differentiation capabilities of healthy MSCs. Dasatinib treatment significantly upregulated the expression of osteogenic and adipogenic-related genes, suggesting that it may enhance the therapeutic potential of MSCs. Additionally, Dasatinib treatment does not impair the immunomodulatory function of MSCs, providing important evidence for its application in regenerative medicine and cell therapy.

Research Highlights

  1. Selective Clearance of Senescent Cells: Dasatinib effectively eliminates senescent MSCs while preserving the functionality of healthy cells.
  2. Enhanced Differentiation Potential: Dasatinib significantly upregulates the expression of osteogenic and adipogenic-related genes, indicating its potential to enhance the therapeutic efficacy of MSCs.
  3. Preserved Immunomodulatory Function: Dasatinib treatment does not compromise the immunosuppressive capacity of MSCs, ensuring its clinical applicability.

Future Research Directions

  1. Telomere Length Analysis: Further investigation into the effects of Dasatinib on telomere length in MSCs to explore its anti-aging mechanisms.
  2. Mitochondrial Function Studies: Research into the effects of Dasatinib on mitochondrial function in MSCs to comprehensively understand its anti-aging effects.
  3. Extracellular Vesicle Secretion: Exploration of the effects of Dasatinib on extracellular vesicle secretion by MSCs to expand its clinical application value.

Research Value

This study provides new strategies for the clinical application of MSCs. By using Dasatinib to clear senescent cells, the therapeutic efficacy of MSCs can be significantly improved. Additionally, the research findings offer important theoretical and experimental evidence for the application of Dasatinib in regenerative medicine and cell therapy, holding significant scientific value and clinical application prospects.