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1. Transcriptome-based screening of ion channels and transporters in a migratory chondroprogenitor cell line isolated from late-stage osteoarthritic cartilage

Author

Matta, C. (Csaba), Lewis, R. (Rebecca), Fellows, C. (Christopher), Diszhazi, G. (Gyula), Almassy, J. (Janos), Miosge, N. (Nicolai), Dixon, J. (James), Uribe, M. C. (Marcos C.), May, S. (Sean), Poliska, S. (Szilard), Barrett-Jolley, R. (Richard), Fodor, J. (Janos), Szentesi, P. (Peter), Hajdú, T. (Tibor), Keller-Pinter, A. (Aniko), Henslee, E. (Erin), Labeed, F. H. (Fatima H.), Hughes, M. P. (Michael P.), and Mobasheri, A. (Ali)

Subjects
osteoarthritis, transcriptomics, chondrocyte, channelome, chondroprogenitor, cartilage, mesenchymal stem cell
Abstract

Chondrogenic progenitor cells (CPCs) may be used as an alternative source of cells with potentially superior chondrogenic potential compared to mesenchymal stem cells (MSCs), and could be exploited for future regenerative therapies targeting articular cartilage in degenerative diseases such as osteoarthritis (OA). In this study, we hypothesised that CPCs derived from OA cartilage may be characterised by a distinct channelome. First, a global transcriptomic analysis using Affymetrix microarrays was performed. We studied the profiles of those ion channels and transporter families that may be relevant to chondroprogenitor cell physiology. Following validation of the microarray data with quantitative reverse transcription-polymerase chain reaction, we examined the role of calcium-dependent potassium channels in CPCs and observed functional large-conductance calcium-activated potassium (BK) channels involved in the maintenance of the chondroprogenitor phenotype. In line with our very recent results, we found that the KCNMA1 gene was upregulated in CPCs and observed currents that could be attributed to the BK channel. The BK channel inhibitor paxilline significantly inhibited proliferation, increased the expression of the osteogenic transcription factor RUNX2, enhanced the migration parameters, and completely abolished spontaneous Ca2+ events in CPCs. Through characterisation of their channelome we demonstrate that CPCs are a distinct cell population but are highly similar to MSCs in many respects. This study adds key mechanistic data to the in-depth characterisation of CPCs and their phenotype in the context of cartilage regeneration.

Published
2021

2. Transcriptome-based screening of ion channels and transporters in a migratory chondroprogenitor cell line isolated from late-stage osteoarthritic cartilage.

Author

Matta C, Lewis R, Fellows C, Diszhazi G, Almassy J, Miosge N, Dixon J, Uribe MC, May S, Poliska S, Barrett-Jolley R, Fodor J, Szentesi P, Hajdú T, Keller-Pinter A, Henslee E, Labeed FH, Hughes MP, and Mobasheri A

Subjects
Calcium Signaling, Cartilage, Articular drug effects, Cartilage, Articular pathology, Cell Line, Cell Proliferation, Chondrocytes drug effects, Chondrocytes pathology, Core Binding Factor Alpha 1 Subunit genetics, Core Binding Factor Alpha 1 Subunit metabolism, Gene Expression Profiling, Humans, Ion Channels genetics, Large-Conductance Calcium-Activated Potassium Channel alpha Subunits genetics, Large-Conductance Calcium-Activated Potassium Channel alpha Subunits metabolism, Membrane Potentials, Membrane Transport Proteins genetics, Osteoarthritis, Knee genetics, Osteoarthritis, Knee pathology, Potassium Channel Blockers pharmacology, Stem Cells drug effects, Stem Cells pathology, Time Factors, Cartilage, Articular metabolism, Cell Movement drug effects, Chondrocytes metabolism, Ion Channels metabolism, Membrane Transport Proteins metabolism, Osteoarthritis, Knee metabolism, Stem Cells metabolism, Transcriptome
Abstract

Chondrogenic progenitor cells (CPCs) may be used as an alternative source of cells with potentially superior chondrogenic potential compared to mesenchymal stem cells (MSCs), and could be exploited for future regenerative therapies targeting articular cartilage in degenerative diseases such as osteoarthritis (OA). In this study, we hypothesised that CPCs derived from OA cartilage may be characterised by a distinct channelome. First, a global transcriptomic analysis using Affymetrix microarrays was performed. We studied the profiles of those ion channels and transporter families that may be relevant to chondroprogenitor cell physiology. Following validation of the microarray data with quantitative reverse transcription-polymerase chain reaction, we examined the role of calcium-dependent potassium channels in CPCs and observed functional large-conductance calcium-activated potassium (BK) channels involved in the maintenance of the chondroprogenitor phenotype. In line with our very recent results, we found that the KCNMA1 gene was upregulated in CPCs and observed currents that could be attributed to the BK channel. The BK channel inhibitor paxilline significantly inhibited proliferation, increased the expression of the osteogenic transcription factor RUNX2, enhanced the migration parameters, and completely abolished spontaneous Ca 2+ events in CPCs. Through characterisation of their channelome we demonstrate that CPCs are a distinct cell population but are highly similar to MSCs in many respects. This study adds key mechanistic data to the in-depth characterisation of CPCs and their phenotype in the context of cartilage regeneration., (© 2021 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.)

Published
2021
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