Aging Reprograms the Signaling, Metabolic, and Gene Regulatory Dynamics in Murine Bone Marrow-Derived Mesenchymal Stem Cells

Bone marrow-derived mesenchymal stem cells (BMSCs), owing to their osteoblastogenic differentiation potential, are crucial for maintaining bone homeostasis and remodeling. Nevertheless, in aging and age-related bone diseases like osteoporosis, BMSCs show significantly diminished osteogenic potential, with a concomitant increase in adipogenic differentiation. The aged BMSCs also become desensitized to BMP2 stimulation to a large extent and exhibit aberrations in BMP2 signaling. However, the molecular mechanisms facilitating this shift in lineage commitment and mediating the cellular dysfunctions remain elusive. This knowledge gap hinders the development of regenerative strategies for skeletal aging and osteoporosis. This study employed an integrative tandem mass tag (TMT)-based phosphoproteomic and total proteomic profiling on BMSCs isolated from young (6-month) and aged (15-month) C57BL/6 (B6) mice to elucidate global alterations in both protein activity and expression. The analysis identified more than 500 proteins that underwent significant alterations (BH-adjusted p-value <0.05) either in phosphorylation or expression between young and aged BMSCs. Many lineage-specific markers also underwent changes in both phosphorylation and expression with aging. Additionally, key biological processes, including cellular metabolism, clathrin-mediated endocytosis, and nucleocytoplasmic transport mechanisms, were enriched for the deregulated proteins. Signaling proteins, ERK-1/2, had increased activating phosphorylation in the aged BMSCs, while transcription factors Lrrfip1, Ruvbl1, and Ruvbl2 also exhibited dysregulated activity and abundance in the aged BMSCs. The findings from the study adds significant mechanistic insights into how aging disrupts signal transduction, metabolism, and transcriptional program in BMSCs, contributing to age-associated loss of bone mass and reduced skeletal regenerative capabilities. Through the identification of key mediators of BMSC dysfunction seen in aging, this work offers a strong foundation in devising potential therapeutic strategies to restore diminished osteogenic potential and treat osteoporosis.