Inverse genetics tracing the differentiation pathway of human chondrocytes
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ABSTRACT:
PROVIDER: PRJNA1089120 | ENA |
REPOSITORIES: ENA
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Project description:Objective: Mammalian somatic cells can be reprogrammed into induced pluripotent stem cells (iPSCs) via the forced expression of Yamanaka reprogramming factors. However, only a limited population of the cells that pass through a particular pathway can metamorphose into iPSCs, while the others do not. This study aimed to clarify the pathways that chondrocytes follow during the reprogramming process. Design: The fate of human articular chondrocytes under reprogramming was investigated through a time-coursed single-cell transcriptomic analysis, which we termed an inverse genetic approach. The iPS interference technique was also employed to verify that chondrocytes inversely return to pluripotency following the proper differentiation pathway . Results: We confirmed that human chondrocytes could be converted into cells with an iPSC phenotype. Moreover, it was clarified that a limited population that underwent the silencing of SOX9, a master gene for chondrogenesis, at a specific point during the proper transcriptome transition pathway, could eventually become iPS cells. Interestingly, the other cells, which failed to be reprogrammed, followed a distinct pathway toward cells with a surface zone chondrocyte phenotype. The critical involvement of cellular communication network factors (CCNs) in this process was indicated. The idea that chondrocytes, when reprogrammed into iPSCs, follow the differentiation pathway backward was supported by the successful iPS interference using SOX9. Conclusions: This inverse genetic strategy is expected to be utilized to identify the master genes for the differentiation of various somatic cells. The utility of CCNs in articular cartilage regeneration is also supported.
2024-06-27 | GSE261806 | GEO
Project description:The mannose-6-phosphate (M6P) pathway is critical for lysosome biogenesis, facilitating the trafficking of hydrolases to lysosomes to ensure cellular degradative capacity [1], [2]. Fibroblast Growth Factor (FGF) signaling, a key regulator of skeletogenesis [3], has been linked to the autophagy-lysosomal pathway in chondrocytes [4], [5], [6], but its role in lysosome biogenesis remains poorly characterized. Here, using mass spectrometry, lysosome immune-purification, and functional assays, we reveal that chondrocytes lacking FGF receptors 3 and 4 exhibit dysregulations of the M6P pathway, resulting in hypersecretion of lysosomal enzymes and impaired lysosomal function. We found that FGF receptors control the expression of M6P receptors genes in response to FGF stimulation and during cell cycle via TFEB/TFE3 activation. Notably, restoring MPRs function—either through gene expression or activation of TFEB—rescues lysosomal defects in FGFR3;4-deficient chondrocytes. These findings uncover a novel mechanism by which FGF signaling regulates lysosomal function, offering insights into the control of chondrocyte catabolism and the understanding of FGF-related human diseases.
2025-08-25 | PXD063031 | Pride
Project description:The mannose-6-phosphate (M6P) pathway is critical for lysosome biogenesis, facilitating the trafficking of hydrolases to lysosomes to ensure cellular degradative capacity [1], [2]. Fibroblast Growth Factor (FGF) signaling, a key regulator of skeletogenesis [3], has been linked to the autophagy-lysosomal pathway in chondrocytes [4], [5], [6], but its role in lysosome biogenesis remains poorly characterized. Here, using mass spectrometry, lysosome immune-purification, and functional assays, we reveal that chondrocytes lacking FGF receptors 3 and 4 exhibit dysregulations of the M6P pathway, resulting in hypersecretion of lysosomal enzymes and impaired lysosomal function. We found that FGF receptors control the expression of M6P receptors genes in response to FGF stimulation and during cell cycle via TFEB/TFE3 activation. Notably, restoring MPRs function—either through gene expression or activation of TFEB—rescues lysosomal defects in FGFR3;4-deficient chondrocytes. These findings uncover a novel mechanism by which FGF signaling regulates lysosomal function, offering insights into the control of chondrocyte catabolism and the understanding of FGF-related human diseases.
2025-08-25 | PXD063029 | Pride
Project description:The mannose-6-phosphate (M6P) pathway is critical for lysosome biogenesis, facilitating the trafficking of hydrolases to lysosomes to ensure cellular degradative capacity [1], [2]. Fibroblast Growth Factor (FGF) signaling, a key regulator of skeletogenesis [3], has been linked to the autophagy-lysosomal pathway in chondrocytes [4], [5], [6], but its role in lysosome biogenesis remains poorly characterized. Here, using mass spectrometry, lysosome immune-purification, and functional assays, we reveal that chondrocytes lacking FGF receptors 3 and 4 exhibit dysregulations of the M6P pathway, resulting in hypersecretion of lysosomal enzymes and impaired lysosomal function. We found that FGF receptors control the expression of M6P receptors genes in response to FGF stimulation and during cell cycle via TFEB/TFE3 activation. Notably, restoring MPRs function—either through gene expression or activation of TFEB—rescues lysosomal defects in FGFR3;4-deficient chondrocytes. These findings uncover a novel mechanism by which FGF signaling regulates lysosomal function, offering insights into the control of chondrocyte catabolism and the understanding of FGF-related human diseases.
2025-08-25 | PXD063036 | Pride
Project description:Inverse Agonists of the Androgen Receptor
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Project description:BBF2H7 (BBF2 human homolog on chromosome 7), an ER-resident basic leucine zipper transcription factor, is activated in response to ER stress and abundantly expresses in chondrocytes. While BBF2H7 is widely expressed in many tissues and organs, the most intense signals were detected in the proliferating zone of the cartilage. We compared gene expressions in primary cultured chondrocytes prepared from rib cartilage between WT and BBF2H7-/- mice at E18.5. Primary cultured chondrocytes were prepared from E18.5 rib cartilage of WT and BBF2H7-/- mice. Chondrocytes were isolated using 0.2% collagenase D (Roche) after adherent connective tissue was removed by 0.2% trypsin (Sigma) and collagenase pretreatment. Isolated chondrocytes were maintained in α-MEM (Gibco) supplemented with 10% FCS and 50 µg/mL ascorbic acid. Adenovirus vectors expressing the mouse p60 BBF2H7 (1-377 aa, BBF-N) were constructed with the AdenoX Expression system (Clontech), according to the manufacturer’s protocol. The cells were infected with adenoviruses 30 h before analysis. We compared gene expressions in primary cultured chondrocytes prepared from rib cartilage between WT and BBF2H7-/- mice at E18.5 using a microarray and various genes associated with protein secretory pathway and ER biogenesis were significantly down-regulated in BBF2H7-/- chondrocytes. We infected primary cultured chondrocytes prepared from BBF2H7-/- mice with adenovirus expressing p60 BBF2H7. Several genes were up-regulated and we picked up them as the direct target of BBF2H7.
2010-05-05 | E-GEOD-18052 | biostudies-arrayexpress
Project description:LMD-RNAseq of resting chondrocytes and proliferative chondrocytes
| PRJNA793434 | ENA
Project description:We investigated the molecular mechanisms that regulate the self-renewal and differentiation of chondroprogenitors by comparing the transcriptome profiles between resting chondrocytes and proliferative chondrocytes
2023-04-26 | GSE192840 | GEO
Project description:Achondroplasia, associated with gain-of-function mutations in FGFR3, causes growth plate cartilage dysfunction, resulting in short-limb dwarfism. However, its precise molecular and cellular mechanisms remain unclear. To address this, we aimed to generate knock-in mice (Fgfr3Ach) harboring the achondroplasia mutation (p.Gly380Arg). In addition to previously reported abnormalities, we observed an expansion of the resting zone. EdU labeling and lineage tracing analysis revealed disrupted stem cell properties of resting zone chondrocytes, preventing self-renewal, limiting cell divisions and migration toward the primary ossification center. Single-cell RNA-seq and immunohistochemical analysis identified a cell cluster that corresponded to the expanded resting zone. Pathway analysis and functional experiments revealed that CREB disrupts stem cell properties in resting zone chondrocytes and contributes to dwarfism. Administration of CREB inhibitor 666-15 restored growth plate pathology and bone length. These findings demonstrate that how excess FGFR3 signaling disrupts resting zone chondrocyte properties and suggest potential therapeutic targets for achondroplasia.
2025-12-21 | GSE275171 | GEO
Project description:The severity of osteoarthritis (OA) and cartilage degeneration are highly correlated with the development of synovitis, which is mediated by the activity of inflammatory macrophages. A better understanding of intercellular communication between inflammatory macrophages and chondrocytes should aid in the discovery of novel therapeutic targets. Here, we explored the pathological role of inflammatory macrophage-extracellular vesicles (EVs) in cartilage degeneration. Macrophages were stimulated by treatment with bacterial lipopolysaccharides to mimic the state of inflammatory macrophages and the resulting EVs (M-LPS EVs) were harvested for chondrocyte stimulation and intraarticular injection in a mouse model. This stimulation resulted in increased catabolism of chondrocytes and cartilage degeneration. Consistently, RNA-seq analyses of stimulated chondrocytes indicated that upregulated genes are mainly categorized into apoptotic process and TNF-signaling pathway which suggests the induction of apoptotic process. These chondrocytes exhibited a significant elevation in the expression of pyroptosis-related molecules that were correlated with the expression of chondrocyte catabolic factors. The disruption of caspase-11 significantly alleviated pyroptotic and catabolic processes in stimulated chondrocytes and the pathological changes in collagenase-induced OA model. Our results provide a new insight into the pathological mechanisms of OA and suggest that non-canonical pyroptosis signaling in chondrocytes represents an attractive therapeutic target for future treatment.
2023-09-20 | GSE199193 | GEO