Osteogenic Differentiation Potential of Mesenchymal Stem Cells Using Single Cell Multiomic Analysis.

Area of Science:

• Stem Cell Biology
• Genomics
• Epigenetics

Background:

• Mesenchymal stem cells (MSCs) are multipotent cells crucial for bone formation.
• Osteoporosis involves reduced osteoblast differentiation and bone formation.
• Donor-derived MSCs exhibit variable osteogenic capacity.

Purpose of the Study:

• To profile the transcriptome and epigenome of MSCs from healthy donors using single-cell multiomics.
• To identify individual-specific regulatory elements and transcriptional regulators involved in osteogenic differentiation.
• To investigate the relationship between MSC regulatory elements and genetic risk for musculoskeletal diseases.

Main Methods:

• Single-cell multiomic analysis (transcriptome and epigenome) of MSCs from four healthy donors.
• Clustering of ~1300-1600 cells per donor to identify distinct cell groups.
• Identification of individual-specific enhancer-promoter pairs and analysis of transcriptional regulator activity.
• Enrichment analysis of genome-wide association study (GWAS) signals for musculoskeletal diseases.

Main Results:

• MSCs from different donors displayed distinct chromatin accessibility and regulatory element landscapes.
• Donor 1 MSCs showed the highest motif activity, predicting greater osteogenic potential.
• Osteoporosis and osteopenia GWAS variants were enriched in regulatory regions of high-potential MSCs (Donor 1), while osteoarthritis variants were linked to Donor 4.
• Bioinformatic predictions of osteogenic potential were validated experimentally.

Conclusions:

• Single-cell multiomics reveals donor-specific regulatory mechanisms governing MSC osteogenic differentiation.
• Distinct regulatory element landscapes in MSCs contribute to variable differentiation potential.
• This approach links genetic risk variants to specific regulatory regions, providing insights into musculoskeletal disease pathogenesis.