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Measuring Single-Cell Aging with an Imaging-based Biomarker of Chromatin and Epigenetic Aging
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Decoding the Regulatory Landscape of Ageing in Musculoskeletal Engineered Tissues Using Genome-Wide DNA Methylation

Mandy Jayne Peffers1, Katarzyna Goljanek-Whysall1, John Collins2

  • 1Institute of Ageing and Chronic Disease, University of Liverpool, Leahurst, Chester High Road, Neston, Wirral, UK, CH64 7TE.

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Summary

Aging impacts mesenchymal stem cell (MSC) differentiation epigenetically. DNA methylation changes affect gene expression in aged MSCs, altering tissue engineering outcomes and offering insights for regenerative medicine in older patients.

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Area of Science:

  • Epigenetics and Regenerative Medicine
  • Stem Cell Biology
  • Aging Research

Background:

  • Mesenchymal stem cells (MSCs) are crucial for regenerative medicine due to their differentiation potential.
  • Epigenetic modifications, particularly DNA methylation, are implicated in cellular aging and gene expression changes.
  • Limited understanding exists regarding the role of DNA methylation in the differentiation of aged MSCs.

Purpose of the Study:

  • To investigate genome-level DNA methylation and gene expression changes in engineered tissues derived from young and aged human MSCs.
  • To identify age-related differential methylation signatures and their impact on chondrogenic, osteogenic, and tenogenic differentiation.
  • To explore the role of alternative splicing and gene ontology in age-related epigenetic alterations.

Main Methods:

  • Genome-wide DNA methylation analysis using Illumina HumanMethylation 450 Beadchip arrays.
  • RNA sequencing to compare gene expression profiles between young and aged MSC-derived tissues.
  • Analysis of differential methylation, gene expression, alternative splicing, and gene ontology.

Main Results:

  • Identified unique and common DNA methylation signatures associated with aging in engineered tissues.
  • Discovered age-related differentially expressed genes predominantly involved in 'cell death and survival,' 'cell morphology,' and 'cell growth and proliferation.'
  • Found dysregulated alternative splicing in aged MSCs, particularly in genes related to metabolic processes, and enrichment in 'skeletal system morphogenesis' and 'regulation of cell proliferation.'

Conclusions:

  • Aging induces significant epigenetic alterations, including DNA methylation and alternative splicing changes, in MSC-derived engineered tissues.
  • These age-related epigenetic modifications impact key cellular pathways and contribute to an altered tissue phenotype.
  • Findings provide novel insights into the aging process in MSCs, with implications for optimizing stem cell therapies in elderly patients.