Transcriptomic and proteomic analysis of signature molecules predictive of chrondrogenic potency and tissue specipicity in human mesenchymal stem cells

Research output: Contribution to journalAbstract

Abstract

Purpose: Osteoarthritis (OA) is a progressive degenerative disorder affecting several compartments of the joints, including articular cartilage. Cell therapy with chondrocytes or mesenchymal stem cells (MSCs) to repair focal lesions of the joint surface, a known risk factor for OA, has been intensely pursued. Although studies have demonstrated improvements in symptomatic outcome and cartilage repair, patient-to-patient variability in structural outcome has been reported. To enhance consistency, cell therapy requires a better insight into MSC signature molecules to select qualifying donors and compliant cell sources for articular cartilage repair with MSCs. In this study we aimed to analyse human MSC transcriptomic and proteomic profiles to identify tissue-specific molecular signatures and biomarkers predictive of their chondrogenic potency.

Methods: Human MSCs from three different joint tissues (human bone marrow = HBM, human periosteum = HP, human synovial membrane = HSM) were culture-expanded. In vitro chondrogenesis assays were performed to select samples with high or low chondrogenic differentiation capacity based on their COL2A1/COL1A1 ratio. Microarrays and liquid chromatography-mass spectrometry (LC-MS) were carried out to compare the transcriptome and proteome profiles of MSCs with high vs. low chondrogenic potency and across the three different tissue sources. Expression of selected candidate genes/proteins was validated by q-RT-PCR and Western blot.

Results: Analysis of the microarray and proteomic datasets revealed significant differences in gene expression profiles between MSCs from high and low chondrogenic potency groups and also across the three tissue sources. The high potency group had highly enriched signature molecules with many important biological functions (e.g., gene expression, protein translation, protein phosphorylation, protein transportation, cell division, cytoskeleton organization, intracellular signal transduction etc.). HSM and BM-MSCs expressed distinct sets of genes. A total of 347 genes and 47 pathways, including pathways involved in skeletogenesis, were differentially expressed. Western blot and proteomic analysis confirmed a unique tissue-specific molecular signature for HSM-MSCs.

Conclusions: Our findings demonstrate that MSCs with relatively high chondrogenic differentiation capacity exhibit a distinct genomic and proteomic profile. We have also demonstrated that HSM and BM-MSCs express distinct sets of genes. Gene lists could be potential molecular signatures for tissue-specific MSC identity in clinical applications.
Original languageEnglish
Article number876
Pages (from-to)S509
Number of pages1
JournalOsteoarthritis and Cartilage
Volume24
Issue numberSuppl. 1
Early online date20 Mar 2016
DOIs
Publication statusPublished - Apr 2016
EventWorld Congress of the Osteoarthritis-Research-Society-International (OARSI) on Osteoarthritis - Amsterdam, Netherlands
Duration: 31 Mar 20163 Apr 2016

Cite this

@article{3075e821f44c46fbb0bb1332748ef428,
title = "Transcriptomic and proteomic analysis of signature molecules predictive of chrondrogenic potency and tissue specipicity in human mesenchymal stem cells",
abstract = "Purpose: Osteoarthritis (OA) is a progressive degenerative disorder affecting several compartments of the joints, including articular cartilage. Cell therapy with chondrocytes or mesenchymal stem cells (MSCs) to repair focal lesions of the joint surface, a known risk factor for OA, has been intensely pursued. Although studies have demonstrated improvements in symptomatic outcome and cartilage repair, patient-to-patient variability in structural outcome has been reported. To enhance consistency, cell therapy requires a better insight into MSC signature molecules to select qualifying donors and compliant cell sources for articular cartilage repair with MSCs. In this study we aimed to analyse human MSC transcriptomic and proteomic profiles to identify tissue-specific molecular signatures and biomarkers predictive of their chondrogenic potency.Methods: Human MSCs from three different joint tissues (human bone marrow = HBM, human periosteum = HP, human synovial membrane = HSM) were culture-expanded. In vitro chondrogenesis assays were performed to select samples with high or low chondrogenic differentiation capacity based on their COL2A1/COL1A1 ratio. Microarrays and liquid chromatography-mass spectrometry (LC-MS) were carried out to compare the transcriptome and proteome profiles of MSCs with high vs. low chondrogenic potency and across the three different tissue sources. Expression of selected candidate genes/proteins was validated by q-RT-PCR and Western blot.Results: Analysis of the microarray and proteomic datasets revealed significant differences in gene expression profiles between MSCs from high and low chondrogenic potency groups and also across the three tissue sources. The high potency group had highly enriched signature molecules with many important biological functions (e.g., gene expression, protein translation, protein phosphorylation, protein transportation, cell division, cytoskeleton organization, intracellular signal transduction etc.). HSM and BM-MSCs expressed distinct sets of genes. A total of 347 genes and 47 pathways, including pathways involved in skeletogenesis, were differentially expressed. Western blot and proteomic analysis confirmed a unique tissue-specific molecular signature for HSM-MSCs.Conclusions: Our findings demonstrate that MSCs with relatively high chondrogenic differentiation capacity exhibit a distinct genomic and proteomic profile. We have also demonstrated that HSM and BM-MSCs express distinct sets of genes. Gene lists could be potential molecular signatures for tissue-specific MSC identity in clinical applications.",
author = "S. Shahana and A. Roelofs and {De Bari}, C.",
year = "2016",
month = "4",
doi = "10.1016/j.joca.2016.01.930",
language = "English",
volume = "24",
pages = "S509",
journal = "Osteoarthritis and Cartilage",
issn = "1063-4584",
publisher = "ELSEVIER APPL SCI PUBL LTD",
number = "Suppl. 1",

}

TY - JOUR

T1 - Transcriptomic and proteomic analysis of signature molecules predictive of chrondrogenic potency and tissue specipicity in human mesenchymal stem cells

AU - Shahana, S.

AU - Roelofs, A.

AU - De Bari, C.

PY - 2016/4

Y1 - 2016/4

N2 - Purpose: Osteoarthritis (OA) is a progressive degenerative disorder affecting several compartments of the joints, including articular cartilage. Cell therapy with chondrocytes or mesenchymal stem cells (MSCs) to repair focal lesions of the joint surface, a known risk factor for OA, has been intensely pursued. Although studies have demonstrated improvements in symptomatic outcome and cartilage repair, patient-to-patient variability in structural outcome has been reported. To enhance consistency, cell therapy requires a better insight into MSC signature molecules to select qualifying donors and compliant cell sources for articular cartilage repair with MSCs. In this study we aimed to analyse human MSC transcriptomic and proteomic profiles to identify tissue-specific molecular signatures and biomarkers predictive of their chondrogenic potency.Methods: Human MSCs from three different joint tissues (human bone marrow = HBM, human periosteum = HP, human synovial membrane = HSM) were culture-expanded. In vitro chondrogenesis assays were performed to select samples with high or low chondrogenic differentiation capacity based on their COL2A1/COL1A1 ratio. Microarrays and liquid chromatography-mass spectrometry (LC-MS) were carried out to compare the transcriptome and proteome profiles of MSCs with high vs. low chondrogenic potency and across the three different tissue sources. Expression of selected candidate genes/proteins was validated by q-RT-PCR and Western blot.Results: Analysis of the microarray and proteomic datasets revealed significant differences in gene expression profiles between MSCs from high and low chondrogenic potency groups and also across the three tissue sources. The high potency group had highly enriched signature molecules with many important biological functions (e.g., gene expression, protein translation, protein phosphorylation, protein transportation, cell division, cytoskeleton organization, intracellular signal transduction etc.). HSM and BM-MSCs expressed distinct sets of genes. A total of 347 genes and 47 pathways, including pathways involved in skeletogenesis, were differentially expressed. Western blot and proteomic analysis confirmed a unique tissue-specific molecular signature for HSM-MSCs.Conclusions: Our findings demonstrate that MSCs with relatively high chondrogenic differentiation capacity exhibit a distinct genomic and proteomic profile. We have also demonstrated that HSM and BM-MSCs express distinct sets of genes. Gene lists could be potential molecular signatures for tissue-specific MSC identity in clinical applications.

AB - Purpose: Osteoarthritis (OA) is a progressive degenerative disorder affecting several compartments of the joints, including articular cartilage. Cell therapy with chondrocytes or mesenchymal stem cells (MSCs) to repair focal lesions of the joint surface, a known risk factor for OA, has been intensely pursued. Although studies have demonstrated improvements in symptomatic outcome and cartilage repair, patient-to-patient variability in structural outcome has been reported. To enhance consistency, cell therapy requires a better insight into MSC signature molecules to select qualifying donors and compliant cell sources for articular cartilage repair with MSCs. In this study we aimed to analyse human MSC transcriptomic and proteomic profiles to identify tissue-specific molecular signatures and biomarkers predictive of their chondrogenic potency.Methods: Human MSCs from three different joint tissues (human bone marrow = HBM, human periosteum = HP, human synovial membrane = HSM) were culture-expanded. In vitro chondrogenesis assays were performed to select samples with high or low chondrogenic differentiation capacity based on their COL2A1/COL1A1 ratio. Microarrays and liquid chromatography-mass spectrometry (LC-MS) were carried out to compare the transcriptome and proteome profiles of MSCs with high vs. low chondrogenic potency and across the three different tissue sources. Expression of selected candidate genes/proteins was validated by q-RT-PCR and Western blot.Results: Analysis of the microarray and proteomic datasets revealed significant differences in gene expression profiles between MSCs from high and low chondrogenic potency groups and also across the three tissue sources. The high potency group had highly enriched signature molecules with many important biological functions (e.g., gene expression, protein translation, protein phosphorylation, protein transportation, cell division, cytoskeleton organization, intracellular signal transduction etc.). HSM and BM-MSCs expressed distinct sets of genes. A total of 347 genes and 47 pathways, including pathways involved in skeletogenesis, were differentially expressed. Western blot and proteomic analysis confirmed a unique tissue-specific molecular signature for HSM-MSCs.Conclusions: Our findings demonstrate that MSCs with relatively high chondrogenic differentiation capacity exhibit a distinct genomic and proteomic profile. We have also demonstrated that HSM and BM-MSCs express distinct sets of genes. Gene lists could be potential molecular signatures for tissue-specific MSC identity in clinical applications.

U2 - 10.1016/j.joca.2016.01.930

DO - 10.1016/j.joca.2016.01.930

M3 - Abstract

VL - 24

SP - S509

JO - Osteoarthritis and Cartilage

JF - Osteoarthritis and Cartilage

SN - 1063-4584

IS - Suppl. 1

M1 - 876

ER -