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An iPSC-based model of Progressive Pseudorheumatoid Arthropathy of Childhood identifies transcriptional changes in patient-iPSC-specific cartilage tissues

ABSTRACT: Bulk RNA-seq is used to study transcriptomic differences between WISP3-deficient and WISP3-sufficient articular cartilage derived from hPSCs in vitro

ORGANISM(S): Homo sapiens

PROVIDER: GSE230194 | GEO | 2023/08/24

REPOSITORIES: GEO

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Human PSC-derived articular cartilage chondrocytes scRNA-seq

Project description:Objectives Traditional approaches to study Progressive Pseudorheumatoid Arthropathy of Childhood (PPAC) failed to uncover how loss-of-function mutations in Wnt inducible secreted protein 3 (WISP3) cause premature cartilage failure in children. We developed a human induced pluripotent stem cell (hiPSC)-based model of cartilage development to elucidate pathological changes in WISP3-deficient articular cartilage tissues compared to WISP3-sufficient isogenic cartilage tissues. Methods We generated iPSCs from 2 patients with PPAC, and we corrected the disease-causing mutation in one of these lines using CRISPR/Cas9. We also created a WISP3-knockout human embryonic stem cell line. We generated articular cartilage tissues from these hPSCs by established directed differentiation methods. Bioinformatic analyses were used to identify differentially expressed genes (DEGs) and cell type abundances, which were validated by quantitative RT-qPCR and in situ hybridization. Results WISP3-deficient articular cartilage tissues exhibited significantly different transcriptomic profiles and cellular composition compared with their isogenic controls. WISP3-deficient cartilage transcriptomes exhibited enriched biological processes such as TGFb signaling and epithelial to mesenchymal transition. We validated increased expression of TGFb-related genes in WISP3-deficient cartilage, and determined that the abundance of chondrocyte subtypes was altered compared with isogenic controls. Conclusions We developed a robust and reproducible model of iPSC-derived cartilage development to better understand the role of WISP3 in cartilage biology, and the pathology of cartilage failure associated with loss of WISP3 in patients with PPAC. We identified several altered biological processes and signaling pathways in WISP3-deficient cartilage that can be validated in the future with additional patient-derived cell lines, or large animal models.

2023-08-24 | GSE230195 | GEO

Expression data from articular and growth plate cartilage

Project description:The aim of the current study was to identify molecular markers for articular cartilage that can be used for the quality control of tissue engineered cartilage. Therefore a genom-wide expression analysis was performed using RNA isolated from articular and growth plate cartilage, both extracted from the knee joints of minipigs. Keywords: Native material or primary cells isolated from articular cartilage and growth plate cartilage Articular and growth plate cartilage were taken for RNA extraction and hybridization on Affymetrix microarrays. Furthermore chondrocytes from each type of cartilage were isolated and cell culture was started and terminated at day 10 or day 20. Total RNA from cultivated cells was extracted, and hybridization on Affymetrix microarrays was performed.

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Effect of different hydrostatic pressures on mouse metatarsal and hip cartilage tissues

Project description:Chondrocytes are subject to continuous loads placed upon them throughout development and physical activity. Normal physiological loads enable the maintenance of the articular cartilage health, however abnormal loads contribute to pathological joint ageing. Similarly, the growth plate cartilage is exposed to a number of loads during growth and development. Due to the high-water content of cartilage, hydrostatic pressure is considered one of the main biomechanical influences on chondrocytes and it plays an important role in the mechanoregulation of cartilage. Therefore in this study we conducted RNA-seq analysis of ex vivo hip cap (articular) and metatarsal (growth) cartilage cultures after physiological and injurious hydrostatic pressure. Gene expression in response to 5mPa (physiological) or 50mPa (injurious) hydrostatic pressure was quantified by transcriptome analysis using the Illumina platform

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Expression data from articular and growth plate cartilage

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Differential Gene Expression Associated With Postnatal Articular Cartilage Maturation

Project description:Chondrocyte gene expression was analyzed to study mechanisms involved in the structural and functional adaptation of articular cartilage during postnatal maturation. Transcriptional profiling was used to compare articular chondrocytes between four neonatal and four adult horses. Expressional differences featured matrix proteins and matrix-modifying enzymes reflecting the transition from cartilage growth to cartilage homeostasis. Keywords: articular cartilage, maturation, horse, cDNA microarray

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Engraftment of allogeneic iPS cell-derived cartilage organoid in a primate model of articular cartilage defect

Project description:Induced pluripotent stem cells (iPSCs) are a promising resource for allogeneic cartilage transplantation to treat articular cartilage defects that do not heal spontaneously and often progress to debilitating conditions, such as osteoarthritis. However, to the best of our knowledge, allogeneic cartilage transplantation into primate models has never been assessed. Here, we show that allogeneic iPSC-derived cartilage organoids survive and integrate as well as function as articular cartilage in a primate model of chondral defects in the knee joints. Histological analysis revealed that allogeneic iPSC-derived cartilage organoids in chondral defects elicited no immune reaction and directly contributed to the tissue repair for at least four months. iPSC-derived cartilage organoids integrated with the host native articular cartilage and prevented degeneration of the surrounding cartilage. Single-cell RNA-sequence analysis indicated that iPSC-derived cartilage organoids differentiated after transplantation, acquiring expression of PRG4 that is crucial for joint lubrication. Pathway analysis suggested the involvement of SIK3 inactivation, verified through molecular experiments. Our study outcomes suggest that allogeneic transplantation of iPSC-derived cartilage organoids may be clinically applicable for the treatment of patients with chondral defects of the articular cartilage.

2023-01-23 | GSE206120 | GEO

Comparison of Transient + articular cartilage (TC+AC) over pure transient cartilage (TC) from both E12 and E14 chicken tibia

Project description:During the past two decades although many genes e.g.,Gdf5, Wnt9a, Noggin etc. have been identified and characterized in joint development, still a comprehensive understanding of molecular network operational in articular cartilage morphogenesis is far from being drawn. This might be due to incompleteness in the number of molecules identified. We used microarray profiling to identify chicken articular cartilage specific genes. We decided to conduct microarray-based transcriptome comparison of HH38 (E12) and HH40 (E14). As it is difficult to clearly dissect out articular cartilage even at these late stages we decided to identify the transcripts that are enriched in the transient + articular cartilage (TC+AC) over pure transient cartilage (TC) thus using TC as control or reference samples

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An iPSC-based model of Progressive Pseudorheumatoid Arthropathy of Childhood identifies transcriptional changes in patient-iPSC-specific cartilage tissues

Project description:An iPSC-based model of Progressive Pseudorheumatoid Arthropathy of Childhood identifies transcriptional changes in patient-iPSC-specific cartilage tissues

| PRJNA957811 | ENA

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