resumo
Mesenchymal stem cells (MSCs) secretion/consumption profile significantly determines their therapeutic effects (e.g., bone regeneration)[1]. However, several extracellular constituents (including metabolites) and their role in osteodifferentiation remain underexplored.[2] Nuclear Magnetic Resonance (NMR)-exometabolomics may reveal new insights into MSCs osteodifferentiation[2,3], elucidate how the exometabolome reflects intracellular behavior, and provide non-invasive differentiation monitoring. Here, untargeted NMR-metabolomics was employed to monitor exometabolome changes of human adipose-derived MSCs from 3 different donors throughout 21 days of osteodifferentiation, compared to proliferation. For all donors, both conditions involved i) consumption of glucose, glutamine, pyruvate, serine, and branched-chain amino acids (BCAAs), and ii) secretion of energy-related compounds (Krebs cycle intermediates, creatine, phosphocreatine and lactate), BCAAs breakdown products, ornithine and histidine. During osteodifferentiation, cells exhibited significant changes in metabolic activity, increasing lactate, ornithine and pyroglutamate secretion, while reducing the consumption of several amino acids, pyruvate and glucose. This suggests glycolytic downregulation, with redirection of pyruvate towards lactate, instead of oxidation through Krebs cycle. Articulating extracellular and intracellular changes provided deeper insight into the suggested hallmarks of osteogenesis, namely adaptations in energy metabolism, cell membrane remodeling, and antioxidative protection. The donor-independent features may be exploited to monitor and predict MSC osteogenic ability and advance potential osteoinductive metabolites for osteocommitment enhancement. Keywords: Osteogenic Differentiation; NMR metabolomics; Exometabolomics; Non-Invasive monitoring.
autores
Bispo, DSC; Rodrigues, JA; Graça, ICR; Correia, M; Jesus, CSH; Oliveira, MB; Mano, JF; Gil, AM
nossos autores
Inês C. R. Graça
Bolseiro de investigaçãoProjectos
Metabolite-activated 3D stem cell differentiation into bone (BetterBone)
A Metabolomics-guided Bioreactor for Improved Engineered Bone Implants (BioImplant)
CICECO - Aveiro Institute of Materials (UIDB/50011/2020)
CICECO - Aveiro Institute of Materials (UIDP/50011/2020)
Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)
agradecimentos
BetterBone (2022.04286.PTDC, DOI 10.54499/2022.04286.PTDC) & BIOIMPLANT (PTDC/BTM-ORG/28835/2017), through COMPETE2020/FEDER (POCI-01-0145-FEDER-028835); CICECO-Aveiro Institute of Materials, UIDB/50011/2020 (DOI 10.54499/UIDB/50011/2020), UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020), FCT/MCTES (PIDDAC); SFRH/BD/150655/2020 (DOI 10.54499/SFRH/BD/150655/2020, DSB PhD grant); National NMR Network (Project Nº022161) through FEDER (COMPETE 2020/POCI/PORL/FCT (PIDDAC).