resumo
Introduction: The secretion profile of mesenchymal stem cells (MSCs) is a major driver of their therapeutic effects, e.g. in bone regeneration. However, some secretome constituents, namely metabolites, are still largely unexplored. In this context, Nuclear Magnetic Resonance (NMR) exometabolomics may reveal new information on MSCs osteodifferentiation and clarify how the secretome reflects intracellular behavior, providing means to monitor differentiation. Methodology: Untargeted NMR metabolomics was applied to monitor exometabolome changes of human adipose-derived MSCs (hAMSCs) from 3 different donors throughout 21 days of osteodifferentiation, compared to proliferation alone (controls). Results: For all donors, both osteogenesis and proliferation involved i) consumption of glucose, glutamine, pyruvate, serine, and branched-chain amino acids (BCAAs), and ii) secretion of several energy-related compounds (e.g. Krebs cycle intermediates, creatine, phosphocreatine and lactate), BCAAs breakdown products, ornithine and histidine. During osteodifferentiation, cells underwent evident changes in metabolic activity, increasing serine and cysteine uptake, while reducing the excretion of histidine (mediates mineralization), creatine and phosphocreatine. Ornithine was highly secreted, most likely to avoid polyamines accumulation and subsequent osteodifferentiation inhibition. Simultaneously, there was less consumption of glutamine, BCAAs and glucose, suggesting glycolytic downregulation with redirection of pyruvate towards reduction to lactate. Exometabolome results were articulated with endometabolome changes in the same cells, providing further insight into the above-mentioned energy metabolic switch, as well as cell membrane metabolism, antioxidative protection and creatine biosynthesis. Conclusion: The donor-independent features unveiled in this work may be exploited to monitor cell osteogenic ability and advance potential metabolic candidates for media optimization towards osteocommitment enhancement.
autores
Daniela S.C. Bispo, Marlene Correia, Joao A. Rodrigues, Mariana B. Oliveira, Joao F. Mano, Ana M. Gil
nossos autores
Ana Maria Pissarra Coelho Gil
Professor Associado com Agregação
Daniela Bispo
Estudante de doutoramento
João A. Rodrigues
Investigador Júnior
João Mano
Professor Catedrático
Mariana Braga de Oliveira
Investigador Auxiliar
Marlene Correia
Estudante de doutoramentoProjectos
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) & 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), through FCT/MCTES (PIDDAC); SFRH/BD/150655/2020 (DSB PhD grant); National NMR Network (Project Nº022161) through FEDER (COMPETE 2020/POCI/PORL/FCT (PIDDAC).