Dataset Information

Comparative proteomic and metabolomic analysis of human osteoblasts differentiated from dental pulp stem cells

ABSTRACT: The current global situation points to the trend of the aging population. Along with the growth of population diseases, the pressure to develop new cell-based or drug-based therapies is increasing. One from main challenges in the field of regenerative medicine is large bones healing and osteoporosis, mainly postmenopausal women, treatment. To understand molecular processes which underlying bone development and turnover, osteogenic differentiation using specific media was applied. Differentiation process imitating real osteogenesis is associated with proteome and metabolome changes resulting in various signaling pathways activation. Proteome was profiled by ultrahigh-performance liquid chromatography and comprehensively quantified by ion mobility enhanced mass spectrometry. From 2267 reproducibly quantified proteins, 432 were differentially abundant by strict statistic and effect size criteria. Metabolome profiling was carried out nuclear magnetic resonance in resulting 27 reproducibly quantified metabolites. From them, 8 were statistically significant and differentially abundant. KEGG and MetaboAnalyst analysis indicated several metabolic pathways involved in osteogenic process. Enrichment analysis of differentialy abundant proteins reported PPAR, FoxO, IL-7 signaling pathways, thyroid hormones and steroid biosynthesis, mineral absorption, JAK-STAT and fatty acid degradation pathways as cascades with prominent impact to osteoinduction. Among them, enrichment analysis of significant metabolites showed Aminoacyl-tRNA biosynthesis as well as Valine, leucine, isoleucine metabolism together with phenylalanine, tyrosine and tryptophane metabolism as pathways which promotes osteodifferentiation. Moreover, we also selected few identified pathways which do not reported enrichment score, but have been described in the literature as majority. These cascades underline the complexity of the whole potential osteogenic mechanism. We also proposed promising targets for next validation in patient samples, what is step forward to influence of bone and/or oral defects.

INSTRUMENT(S): SYNAPT G2-Si

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Stem Cell, Dental Pulp

DISEASE(S): Disease Free

SUBMITTER: Maksym Danchenko

LAB HEAD: Andrej Kováč

PROVIDER: PXD025223 | Pride | 2021-09-10

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	checksum.txt	Txt
	sdrf-stem-cell.tsv	Tabular
	stem-cell.DerivedFromSearchf63c158c-7869-4037-8250-d2bb47be4623.mgf	Mgf
	stem-cell.mzid.gz	Mzid
	stem-cell.zip	Other

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Publications

Comparative Proteomic and Metabolomic Analysis of Human Osteoblasts, Differentiated from Dental Pulp Stem Cells, Hinted Crucial Signaling Pathways Promoting Osteogenesis.

Nováková Slavomíra S Danchenko Maksym M Okajčeková Terézia T Baranovičová Eva E Kováč Andrej A Grendár Marián M Beke Gábor G Pálešová Janka J Strnádel Ján J Janíčková Mária M Halašová Erika E Škovierová Henrieta H

International journal of molecular sciences 20210724 15

Population aging has been a global trend for the last decades, which increases the pressure to develop new cell-based or drug-based therapies, including those that may cure bone diseases. To understand molecular processes that underlie bone development and turnover, we followed osteogenic differentiation of human dental pulp stem cells (DPSCs) using a specific induction medium. The differentiation process imitating in vivo osteogenesis is triggered by various signaling pathways and is associated ...[more]

PMID: 34360674

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Project description:The treatment of bone defects caused by infection, trauma or neoplasms remains a clinical challenge. Autologous bone transplantation is limited by availability, donor site morbidity and surgical risk factors. This has given rise to stromal/stem-cell based therapy. Bone marrow derived stromal cells (BMSCs) have been studied to a large extent and show high regenerative potential but their use is limited by availability, donor site morbidity and the relatively low cell yield as they represent only <0.1% of cell harvested from bone marrow aspirate. At the same time, they are the closest mesenchymal stromal cells for bone tissue engineering given their tissue origin and, unlike other mesenchymal stromal cells, can support the formation of hematopoietic marrow. Adipose tissue derived stromal cells (ASCs) as part of the stromal vascular fraction of adipose tissue can as well undergo osteogenic differentiation but can be additionally isolated in a sufficient quantity from lipoaspirate after liposuction of abundant subcutaneous fat tissue. Here, it has been shown that there are no major differences in regard to proliferation or differentiation capacity of ASCs derived from subcutaneous fat of different anatomical regions. It has been shown that BMSCs are more prone to senescence during expansion and passage than ASCs and that ageing impacts proliferative capabilities of BMSCs more than that of ASCs while it has also been reported that osteogenic differentiation capacity is least impacted by age. Multiple studies have compared the characteristics of these two mesenchymal stromal cells in regard to bone tissue engineering in vitro. Most studies point to inferior extracellular matrix mineralization and lower expression of key osteogenic transcription markers like Runx2 in osteogenic differentiated ASCs compared to BMSCs. On the other hand, a study by Rath et al. found contrary results using particular culturing conditions like 3D bioglass scaffolds. An intraindividual comparison of human MSCs of three donors cultured on decellularized porcine bone confirmed superior osteogenic capacity of BMSCs compared to ASCs. In contrast to BMSCs, ASCs were not able to induce heterogenic ossification in a mouse model. In a sheep tibia defect model application of BMSCs resulted in a significantly higher amount of newly formed bone tissue. Importantly, Osteogenic differentiated ASCs do not support the formation of a hematopoietic marrow. Proteomics enables large-scale analysis of proteins present in a cell type and can be used to identify differentially regulated key proteins in a comparative approach. A comparative proteomic analysis of BMSCs and ASCs by Roche et al. in 2009 identified 556 proteins with 78% of these not being differentially regulated between these two cell populations, regarded as high similarity. Another comparative proteomic study of 2016 by Jeon et al. found 90 differentially regulated proteins out of 3000 total identified proteins. Both studies do not specify a number of different tissue donors and in part using cell lines. Looking for differences upon osteogenic differentiation, transcriptomic comparison of osteogenic differentiated porcine ASCs and BMSCs has been performed, resulting in 21 differentially expressed genes after 21 days of osteogenic culture conditions. Still, it remains unanswered, which are the key distinctive features of osteogenic differentiated ASCs and BMSCs at protein level that might help address the abovementioned weaknesses of ASCs in bone tissue engineering/regeneration for translational research. To overcome this need, an intraindividual comparative DIA based proteomic analysis of osteogenic differentiated human BMSC and ASCs was performed in this study.