Biology subjects

Tuerlings, M.

Publications and source records attributed to Tuerlings, M..

6 recordsLinked to original sources

bioRxiv2026

The Leiden ex vivo human growth plate model in severe tall stature: a proof-of-concept study

Background: Viable human growth plate (GP) tissue is rarely available for translational research, limiting direct investigation of human longitudinal bone growth and pediatric growth disorders. In this proof-of-concept study, we aimed to determine feasibility to establish a clinically integrated ex vivo human growth plate model using tissue obtained during routine percutaneous epiphysiodesis (PE) procedures in adolescents treated for extreme tall stature or leg length difference due to trauma. Methods: Growth plate tissue and cells were collected during PE and processed using protocols adapted from established methods of human osteoarthritic cartilage collection within the RAAK study. Collected tissues and three-dimensional cartilage pellets generated from isolated cells were assessed by histology. Proliferation of GP-derived chondrocytes was compared with osteoarthritis-derived articular chondrocytes. Results: Across consecutive surgical procedures, viable GP tissue could be obtained reproducibly including the node of Ranvier, with only few samples failing to yield cells and no contamination. Isolated GP chondrocytes expanded successfully in two-dimensional culture and showed a stronger early proliferative response compared with RAAK-derived chondrocytes. In addition, GP-derived cells formed three-dimensional organoids and histology confirmed cartilage-like matrix deposition supporting their capacity to generate neo-cartilage tissue in vitro. Conclusion: This study demonstrates feasibility to obtain, culture, and functionally assess viable human growth plate tissue from routine PE surgery. As such, the Leiden ex vivo human growth plate model provides a unique platform to study local mechanisms of endochondral bone growth, link genetic determinants of height to functional growth plate biology, and support future therapeutic research in pediatric growth disorders.

developmental biology↗

bioRxiv2026

Spatial transcriptomics defines the mechanisms of hiPSC derived stem cell mediated repair in human articular cartilage

We here determined therapeutic efficacy and mode-of-action of human induced pluripotent-derived therapeutic stem cells (hiMSCs) across in vivo mouse and ex vivo human osteoarthritis models. hiMSC treatment in DMM-mice significantly reduced OARSI damage scores, which was affirmed by a decrease in the catabolic marker Mmp13 and an increase in the anabolic marker Col2. These treatment effects appeared, irrespective of modifying factors such as xeno-free media or thermosensitive hydrogel carrier. Subsequently treatment of hiMSC+gel in human osteoarthritic cartilage explants showed a transcriptome-wide significant activation of the cholesterol and sterol synthesis pathways marked by genes such as MVD, DHCR7, MSMO1, FABP3. Additionally, we showed that these changes alleviated OA-associated imbalances of the cellular Zinc-ion homeostasis pathways, represented by genes such as MT1F, MT1G, MT1H and SLC30A1. Spatial transcriptomics then sensitively captured that hiMSC+gel treatment evoked, specifically at the superficial cartilage layer, a consistent upregulation of healthy chondrocyte markers such as CHAD, ACAN, FRZB, and SOX9, alongside a suppression of catabolic and inflammatory mediators such as SERPINE1, SPP1, MMP13, ADAMTS5. Our findings link therapeutic outcomes of hiMSC treatment to precise spatially resolved molecular changes in human tissue, that would otherwise be obscured by heterogeneous cell populations. Collectively our study highlighted that hiPSC-derived stem cell therapy (hiMSCs) could provide a scalable off-the-shelf solution to treat osteoarthritis, with strong prospects for clinical applications in the near future.

developmental biology↗

bioRxiv2025

Evaluating the Therapeutic Efficacy of Iopanoic Acid in a DMM-Induced Osteoarthritis Mouse Model and Osteochondral Lesioned Human Explants

ObjectiveTo evaluate the therapeutic potential of iopanoic acid (IOP), a thyroid hormone pathway inhibitor, in preserving cartilage and bone integrity in osteoarthritis (OA), using in vivo and ex vivo tissue models. DesignIn the DMM mouse model, IOP was administered through intra-articular (i.a.) injection, either alone or combined with a thermosensitive hydrogel to enable sustained release. Histological analyses included Safranin O/Fast Green staining and OARSI scoring. Immunohistochemistry was performed for COL2, MMP13, and CCDC80 to evaluate anabolic, catabolic, and hypertrophic markers. Micro-CT assessed subchondral bone changes. In the ex vivo studies, IOP was applied to lesioned human osteochondral OA explants. Matrix degradation and repair were evaluated by sulfated glycosaminoglycan (sGAG) release, Mankin histology scores, and RT-qPCR for cartilage matrix genes. ResultsAdministration of IOP significantly reduced cartilage degeneration in DMM mice (P [≤] 1.0x10-4), characterized by increased COL2, and decreased MMP13 and CCDC80 expression. Notably, IOP also prevented pathological subchondral bone thickening. In human explants, IOP treatment led to a significant reduction in sGAG release compared to untreated explants on day 6 of the IOP treatment. Moreover, Mankin scores were significantly improved in IOP-treated compared to untreated explants, indicating reduced cartilage degradation. ConclusionIOP demonstrates strong chondroprotective effects, reducing cartilage degradation and promoting repair in OA models. Its combination with a thermosensitive hydrogel amplifies therapeutic potential, offering a promising strategy for OA treatment. Next steps are to optimize delivery and validate early molecular effects.

molecular biology↗

bioRxiv2022

WWP2 osteoarthritis risk allele rs1052429-A confers risk by affecting cartilage matrix deposition via hypoxia associated genes

ObjectiveTo explore the co-expression network of the osteoarthritis (OA) risk gene WWP2 in articular cartilage and study cartilage characteristics when mimicking the effect of OA risk allele rs1052429-A on WWP2 expression in a human 3D in vitro model of cartilage. MethodsCo-expression behavior of WWP2 with genes expressed in lesioned OA articular cartilage (N=35 samples) was explored. By applying lentiviral particle mediated WWP2 upregulation in 3D in vitro pellet cultures of human primary chondrocytes (N=8 donors) the effects of upregulation on cartilage matrix deposition was evaluated. Finally, we transfected primary chondrocytes with miR-140 mimics to evaluate whether miR-140 and WWP2 are involved in similar pathways. ResultsUpon performing Spearman correlations in lesioned OA cartilage, 98 highly correlating genes (|{rho}|>0.7) were identified. Among these genes, we identified GJA1, GDF10, STC2, WDR1, and WNK4. Subsequent upregulation of WWP2 on 3D chondrocyte pellet cultures resulted in a decreased expression of COL2A1 and ACAN and an increase in EPAS1 expression. Additionally, we observed a decreased expression of GDF10, STC2, and GJA1. Proteomics analysis identified 42 proteins being differentially expressed with WWP2 upregulation, which were enriched for ubiquitin conjugating enzyme activity. Finally, upregulation of miR-140 in 2D chondrocytes resulted in significant upregulation of WWP2 and WDR1. ConclusionsMimicking the effect of OA risk allele rs1052429-A on WWP2 expression initiates detrimental processes in the cartilage shown by a response in hypoxia associated genes EPAS1,GDF10, and GJA1 and a decrease in anabolic markers, COL2A1 and ACAN.

cell biology↗

bioRxiv2020

Elucidating another level of epigenetic regulation in osteoarthritis by identifying functional cis-acting long non-coding RNAs and their targets in articular cartilage

ObjectiveTo identify robustly differentially expressed long non-coding RNAs (lncRNAs) with osteoarthritis (OA) pathophysiology in cartilage. Moreover to explore potential target mRNAs by establishing co-expression networks, followed by functional validation. MethodsRNA sequencing was performed on macroscopically lesioned and preserved OA cartilage of patients who underwent a joint replacement surgery due to OA (N=98). Differential expression (DE) analysis was performed on lncRNAs that were annotated in GENCODE and Ensembl. To identify potential interactions, correlations were calculated between the identified DE lncRNAs and previously reported DE protein-coding genes in the same samples. Modulation of chondrocyte lncRNA expression was achieved using LNA GapmeRs. ResultsBy applying our in-house pipeline we identified 5,053 lncRNAs to be robustly expressed, of which 191 were FDR significant differentially expressed between lesioned and preserved OA cartilage. Upon integrating mRNA sequencing data, we showed that intergenic and antisense DE lncRNAs show high, positive correlations with their flanking, respectively, sense genes. To functionally validate this observation we selected P3H2-AS1, which was downregulated in primary chondrocytes, resulting in downregulation of P3H2 gene expression levels. As such, we can confirm that P3H2-AS1 regulates its sense gene P3H2. ConclusionBy applying an improved detection strategy, robustly differentially expressed lncRNAs in OA cartilage were detected. Integration of these lncRNAs with differential mRNA expression levels in the same samples showed insight into their regulatory networks. Our data signifies that intergenic, as well as antisense lncRNAs play an important role in regulating the pathophysiology of OA.

genomics↗

bioRxiv2020

RNA sequencing reveals interacting key determinants of osteoarthritis acting in subchondral bone and articular cartilage.

ObjectiveThe aim of this study was to identify key determinants of the interactive osteoarthritis (OA) pathophysiological processes of subchondral bone and cartilage. MethodsWe performed RNA sequencing on macroscopically preserved and lesioned OA subchondral bone of patients that underwent joint replacement surgery due to OA (N=24 pairs; 6 hips, 18 knees, RAAK-study). Unsupervised hierarchical clustering and differential expression analyses were performed. Results were combined with previously identified, differentially expressed genes in cartilage (partly overlapping samples) as well as with recently identified OA risk genes. ResultsWe identified 1569 genes significantly differentially expressed between lesioned and preserved subchondral bone, including CNTNAP2 (FC=2.4, FDR=3.36x10-5) and STMN2 (FC=9.6, FDR=3.36x10-3). Among the identified genes, 305 were also differentially expressed and with same direction of effects in cartilage, including IL11 and CHADL, recently acknowledged OA susceptibility genes. Upon differential expression analysis stratifying for joint site, we identified 509 genes exclusively differentially expressed in subchondral bone of the knee, such as KLF11 and WNT4. These exclusive knee genes were enriched for involvement in epigenetic processes, characterized by for instance HIST1H3J and HIST1H3H. ConclusionTo our knowledge, we are the first to report on differential gene expression patterns of paired OA subchondral bone tissue using RNA sequencing. Among the most consistently differentially expressed genes with OA pathophysiology in both bone and cartilage were IL11 and CHADL. As these genes were recently also identified as robust OA risk genes they classify as attractive druggable targets acting on two OA disease relevant tissues.

genomics↗