Deep phenotyping of an ATDC5 in-vitro cartilage model system
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE324360 | GEO | 2026/06/05
REPOSITORIES: GEO
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Project description:Directed differentiation of stem cells toward chondrogenesis in vitro and in situ to regenerate cartilage suffers from off-target differentiation and hypertrophic tendency. Here, we generated a cartilaginous organoid system from human expanded pluripotent stem cells (hEPSCs) carrying a COL2A1mCherry and COL10A1eGFP double reporter, enabling real-time monitoring of chondrogenesis and hypertrophy. After screening 2,040 FDA-approved drugs, we found that α-adrenergic receptor (α-AR) antagonists, especially phentolamine, stimulated chondrogenesis but repressed hypertrophy, while α2-AR agonists reduced chondrogenesis and induced hypertrophy. Phentolamine prevented cartilage degeneration in hEPSC cartilaginous organoid and human cartilage explant models and stimulated microfracture-activated endogenous skeletal stem cells toward hyaline-like cartilage regeneration without fibrotic degeneration in situ. Mechanistically, α2-AR signaling induced hypertrophic degeneration via cyclic guanosine monophosphate (cGMP)-dependent secretory leukocyte protease inhibitor (SLPI) production. SLPI-deleted cartilaginous organoid was degeneration resistant, facilitating large cartilage defect healing. Ultimately, targeting α2-AR/SLPI was a promising and clinically feasible strategy to regenerate cartilage via promoting chondrogenesis and repressing hypertrophy.
2024-09-30 | GSE271814 | GEO
Project description:Directed differentiation of stem cells toward chondrogenesis in vitro and in situ to regenerate cartilage suffers from off-target differentiation and hypertrophic tendency. Here, we generated a cartilaginous organoid system from human expanded pluripotent stem cells (hEPSCs) carrying a COL2A1mCherry and COL10A1eGFP double reporter, enabling real-time monitoring of chondrogenesis and hypertrophy. After screening 2,040 FDA-approved drugs, we found that α-adrenergic receptor (α-AR) antagonists, especially phentolamine, stimulated chondrogenesis but repressed hypertrophy, while α2-AR agonists reduced chondrogenesis and induced hypertrophy. Phentolamine prevented cartilage degeneration in hEPSC cartilaginous organoid and human cartilage explant models and stimulated microfracture-activated endogenous skeletal stem cells toward hyaline-like cartilage regeneration without fibrotic degeneration in situ. Mechanistically, α2-AR signaling induced hypertrophic degeneration via cyclic guanosine monophosphate (cGMP)-dependent secretory leukocyte protease inhibitor (SLPI) production. SLPI-deleted cartilaginous organoid was degeneration resistant, facilitating large cartilage defect healing. Ultimately, targeting α2-AR/SLPI was a promising and clinically feasible strategy to regenerate cartilage via promoting chondrogenesis and repressing hypertrophy.
2024-09-30 | GSE249229 | GEO
Project description:Deep phenotyping of an ATDC5 in-vitro cartilage model system
| PRJNA1434474 | ENA
Project description:Endochondral ossification is the process by which the appendicular skeleton, facial bones, vertebrae and medial clavicles are formed and relies on the tight control of chondrocyte maturation. Fibroblast growth factor receptor (FGFR)3 plays a role in bone development and maintenance and belongs to a family of proteins which differ in their ligand affinities and tissue distribution. Activating mutations of the FGFR3 gene lead to craniosynostosis and multiple types of skeletal dysplasia with varying degrees of severity: thanatophoric dysplasia (TD), achondroplasia and hypochondroplasia. Despite progress in the characterization of FGFR3-mediated regulation of cartilage development, many aspects remain unclear. The aim and the novelty of our study was to examine whole gene expression differences occurring in primary human chondrocytes isolated from normal cartilage or pathological cartilage from TD-affected fetuses, using Affymetrix technology. The phenotype of the primary cells was confirmed by the high expression of chondrocytic markers. Altered expression of genes associated with many cellular processes was observed, including cell growth and proliferation, cell cycle, cell adhesion, cell motility, metabolic pathways, signal transduction, cell cycle process and cell signaling. Most of the cell cycle process genes were down-regulated and consisted of genes involved in cell cycle progression, DNA biosynthesis, spindle dynamics and cytokinesis. About eight percent of all modulated genes were found to impact extracellular matrix (ECM) structure and turnover, especially glycosaminoglycan (GAG) and proteoglycan biosynthesis and sulfation. Altogether, the gene expression analyses provide new insight into the consequences of FGFR3 mutations in cell cycle regulation, onset of pre-hypertrophic differentiation and concomitant metabolism changes. Moreover, impaired motility and ECM properties may also provide clues about growth plate disorganization. These results also suggest that many signaling pathways may be directly or indirectly altered by FGFR3 and confirm the crucial role of FGFR3 in the control of growth plate development.
2009-12-27 | E-MEXP-2276 | biostudies-arrayexpress
Project description:Owing to the limited regenerative capacity of articular cartilage, damaged cartilage does not spontaneously heal over time. Various scientific efforts have been made to improve damaged articular cartilage. Nevertheless, no single approach has achieved a promising outcome for the damaged cartilage. Meanwhile, owing to the increasing cost of drug development, drug repositioning has been actively attempted. We aimed to identify the drug that can improve the cartilage defect, using chondrogenesis related microarray data recruited from the Gene Expression Omnibus (GEO) public database. Furthermore, we attempted to experiment using cellular and animal models to verify the cartilage regeneration potential for the identified drug. To screen for drugs that promote cartilage restoring, chondrogenesis related microarray data were collected from the GEO public database. The GSE69110, GSE107649, GSE111822, and GSE116173 datasets from the GEO were used to identify cartilage differentiation-related genes. Differentially expressed genes were identified using StringTie, and drug data were extracted from the Drug-Gene Interaction database. The effect of aripiprazole on cartilage was evaluated in aripiprazole-treated adipose-derived mesenchymal stem cells (ADMSCs) and chondrocyte using qRT-PCR and 3D pellet culture. The cartilage restoring efficacy was verified in vivo by mixing it with a scaffold and introducing it into the artificially damaged cartilage of Sprague-Dawley rats. Next, mRNA was sequenced for mechanistic analysis. As a result, aripiprazole significantly increased the mRNA expression of COL2A1 and SOX9, two cartilage differentiation-related genes, and chondrogenic condensation in vitro. Moreover, it effectively promoted cartilage regeneration in the cartilage defect rat model. Analysis of mRNA sequencing data from chondrocyte treated with aripiprazole, using KEGG and GOBP, indicated that aripiprazole significantly upregulates genes associated with ribosomes and cytoplasmic translation, thus promoting chondrogenesis. In conclusion, we discovered that aripiprazole can effectively improve damaged cartilage, providing a promising approach for cartilage regeneration.
2024-12-30 | GSE271026 | GEO
Project description:Long non-coding RNAs (lncRNAs) play pivotal roles in diseases such as osteoarthritis (OA). However, knowledge of the biological roles of lncRNAs is limited in OA. We aimed to explore the biological function and molecular mechanism of HOTTIP in chondrogenesis and cartilage degradation. We used the human mesenchymal stem cell (MSC) model of chondrogenesis, in parallel with, tissue biopsies from normal and OA cartilage to detect HOTTIP, CCL3, and miR-455-3p expression in vitro. Biological interactions between HOTTIP and miR-455-3p were determined by RNA silencing and overexpression in vitro. We evaluated the effect of HOTTIP on chondrogenesis and degeneration, and its regulation of miR-455-3p via competing endogenous RNA (ceRNA). Our in vitro ceRNA findings were further confirmed within animal models in vivo. Mechanisms of ceRNAs were determined by bioinformatic analysis, a luciferase reporter system, RNA pull-down, and RNA immunoprecipitation (RIP) assays. We found reduced miR-455-3p expression and significantly upregulated lncRNA HOTTIP and CCL3 expression in OA cartilage tissues and chondrocytes. The expression of HOTTIP and CCL3 was increased in chondrocytes treated with interleukin-1β (IL-1β) in vitro. Knockdown of HOTTIP promoted cartilage-specific gene expression and suppressed CCL3. Conversely, HOTTIP overexpression reduced cartilage-specific genes and increased CCL3. Notably, HOTTIP negatively regulated miR-455-3p and increased CCL3 levels in human primary chondrocytes. Mechanistic investigations indicated that HOTTIP functioned as ceRNA for miR-455-3p enhanced CCL3 expression. Taken together, the ceRNA regulatory network of HOTTIP/miR-455-3p/CCL3 plays a critical role in OA pathogenesis and suggests HOTTIP is a potential target in OA therapy.
2019-08-12 | PXD014988 | Pride
Project description:Osteoarthritis (OA) is a degenerative joint disease that involves destruction of articular cartilage and eventually leads to disability. Mesenchymal stem cells (MSCs) reside in healthy and diseased cartilage, and through their chondrogenic potential may provide a strategy for cartilage repair. To this end, we performed an image-based, high throughput screen and identified the small molecule, kartogenin, that promotes selective MSC differentiation into chondrocytes (EC50=100nM), shows chondroprotective effects in vitro, and is efficacious in two OA animal models. Kartogenin binds filamin A and induces chondrogenesis by regulating the CBFbeta-RUNX1 transcriptional program. This work provides new insights into the control of chondrogenesis that may ultimately lead to an effective stem-cell based therapy for osteoarthritis. one compound, three doses, two time points, a vehicle control
2012-05-01 | E-GEOD-35546 | biostudies-arrayexpress
Project description:Objective: Skeletal stem cells are known to be regulated by their local self-regulating niche, that is comprised mainly of their progeny and secreted paracrine factors. What cell intrinsic factors govern the maintenance and function of the SSC remains elusive. Methods: Using the previously characterized Tet1-/- mice we investigated the differences in skeletal stem cell lineage tree. Further we examined the ability of these cells to differentiate into osteogenic and chondrogenic lineages, both with and without injury. Transcriptomic differences and inflammatory cytokines were investigated for differences in cartilage development pathways. Results: Loss of Tet1 skews the SSC lineage tree by expanding the SSC pool at the expense of its progeny as well as by enhancing the chondrogenic differentiation of SSC and the multilineage bone cartilage stromal progenitors (BCSP). Tet1 inhibition leads to enhanced chondrogenesis in mouse BCSP upon injury, in human SSC and chondroprogenitors (CP) isolated from osteoarthritic (OA) human cartilage and in late stages of OA in a mouse model. Transcriptomic analyses of SSC and BCSP lacking Tet1 revealed dysregulated pathways including alterations in Tgfb signaling, melatonin degradation and cartilage development associated genes. Conclusion: Tet1 was identified as critical regulator of SSC fate. Loss of Tet1 selectively enhances chondrogenic ability of skeletal stem cells. Supplementation of melatonin can dampen inflammation and lead to better cartilage health.
2023-08-15 | GSE232666 | GEO
Project description:Previously, induced pluripotent stem cells (iPSCs)-derived cartilage have demonstrated superior capacity to regenerate hyaline cartilage compared to mesenchymal stromal cells (MSCs). Since many patients requiring chondral repair are young, the regenerated cartilage must maintain a hyaline cartilage phenotype and function for decades. However, most animal studies testing iPSC chondrogenesis have only conducted short-term assessments. In this study, we first performed a long-term (8 weeks) in vitro chondrogenesis of human iPSC-derived multipotent cells (iMPCs) and human MSCs, and examined cell phenotypes at various time points. The results showed that the expression levels of chondrogenic genes in MSCs peaked earlier than iMPCs. The expression levels of hypertrophy-related genes increased over time in both groups, which however were lower in the iMPC group than the MSC group. Furthermore, iMPC-derived chondrocytes exhibited significantly lower apoptosis levels than MSC-derived chondrocytes. We then implanted the in vitro-generated cartilage into osteochondral defects in normal rats up to 48 weeks (equivalent to ~30 years in humans). At both short and long-term time points (12 and 48 weeks), the cartilage tissue in the defect sites from the iMPC group maintained more cartilage matrix with less collagen type I and osteocalcin deposition than the MSC group. Importantly, at 48 weeks, the native cartilage surrounding the defect areas from the MSC group showed continuous degradation in some rats, which was not observed in the iMPC group. In conclusion, iMPCs represent a safe and effective cell source for long-term hyaline cartilage repair.
2026-01-28 | GSE288164 | GEO
Project description:Histone methylation plays a critical role in cartilage development, making it an appealing target for improving cartilage engineering protocols. In this study, we evaluated the roles of JMJD3 (KDM6B) and UTX (KDM6A), both demethylases of histone H3 at lysine 27 (H3K27), in chondrogenesis and their application in gene-modified mesenchymal stem cell therapy for cartilage tissue engineering. Our findings revealed that JMJD3 is specifically upregulated during chondrogenesis in hBM-MSC, and is crucial for this differentiation process. In contrast, UTX was found to be dispensable for chondrogenesis. Nevertheless, both JMJD3 and UTX share the ability to demethylate similar gene loci, thereby promoting the expression of chondrogenic signature genes, which suggests functional redundancy. Notably, the genes encoding these H3K27me3 demethylases emerge as strong candidates for enhancing gene-modified mesenchymal stem cell therapy for cartilage tissue engineering, as their overexpression during chondrogenesis significantly increased the formation of thicker cartilage discs enriched with type II collagen. These findings emphasize the potential of transient JMJD3 transduction, along with a lesser emphasis on UTX, as effective strategies for improving gene-modified mesenchymal stem cell therapy in cartilage tissue engineering.
2025-10-01 | GSE303471 | GEO