
Human cartilage model of the precocious osteoarthritis-inducing COL2A1 p.R719C reveals pathology-driving matrix defects and a failure of the ER proteostasis network to recognize the defective procollagen-II

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Kathryn M. Yammine, Sophia Mirda Abularach, Michael Xiong, Seo-yeon Kim, Agata A. Bikovtseva, Vincent L. Butty, Richard P. Schiavoni, John F. Bateman, Shireen R. Lamandé, and Matthew D. Shoulders
A dbGaP Submission is underway and metadata will be updated at the completion of submission. The remaining of the data and metadata are available here.
DOI: 10.1101/2024.11.07.622468.
Abstract:
Objectives Mutations in the procollagen-II gene (COL2A1) often cause chondrodysplasias, including the precocious osteoarthritis-inducing p.Arg719Cys. Understanding the molecular basis of such diseases has long been challenging, owing to a lack of models accurately reflecting disease genotypes and phenotypes. To address this challenge, we develop and characterize in vitro human cartilage derived from wild-type and disease-causing Arg719Cys COL2A1 isogenic induced pluripotent stem cell (iPSC) lines.
Methods Using directed differentiation of iPSCs to chondrocytes, we generated cartilage from wild-type and Arg719Cys COL2A1 lines. We compared the resulting protein, cell, and tissue properties using immunohistochemistry, electron microscopy, SDS-PAGE, RNA-sequencing, and quantitative interactomics.
Results While both wild-type and disease lines deposited a cartilage matrix, the Arg719Cys matrix was deficient. Arg719Cys collagen-II was excessively post-translationally modified and modestly intracellularly retained, leading to endoplasmic reticulum (ER) distention suggestive of an ER storage defect. Interactomic studies indicated that Arg719Cys procollagen-II was not differentially engaged by the ER proteostasis network. RNA-sequencing showed that the ER storage defect engendered by Arg719Cys procollagen-II also did not activate cellular stress responses, including the unfolded protein response. These data suggest that cells fail to properly recognize Arg719Cys-associated procollagen-II defects.
Conclusions A failure to identify and rectify defective procollagen-II folding in cells expressing Arg719Cys procollagen-II leads to the deposition of a sparse and defective collagen-II matrix, culminating in pathology. Combined with the highly expandable human cartilage disease model reported here, this work provides motivation and a platform to discover therapeutic strategies targeting procollagen folding, quality control, and secretion in this collagenopathy and others.
SEEK ID: https://fairdomhub.org/studies/1368
Shoulders Lab Individual Publications
Projects: Shoulders Lab
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Created: 3rd Jun 2025 at 15:41
Last updated: 9th Jun 2025 at 14:07

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Projects: Hi-IMPAcTB, MIT SRP, MetNet, Endometriosis, Cancer Systems Biology Consortium (CSBC), Shoulders Lab
Institutions: Massachusetts Institute of Technology
Projects: MIT SRP, Hi-IMPAcTB, MetNet, Endometriosis, Cancer Systems Biology Consortium (CSBC), Shoulders Lab
Web page: https://nextseek.mit.edu/
Shoulders Lab Collagen Publications
Programme: NExtSEEK
Public web page: Not specified
Organisms: Not specified
Submitter: Charles Demurjian
Studies: Human cartilage model of the precocious osteoarthritis-inducing COL2A1 p..., In vivo directed evolution of an ultrafast Rubisco from a semianaerobic ...
Assays: Cell Imaging - Data Linked, Gene Expression Analysis - Data Linked, LC - MS/MS (Proteomics) - Data Linked, Library Creation - Metadata, Proteomics Analysis - Data Linked, RNA Extraction - Metadata, Short Read Sequencing - Data Linked, iPSC Culture and Chondronoid Formation - Metadata
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Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_Imaging.docx, P.SHO-250605-V1_Collagen_Proteomics.docx, P.SHO-250605-V1_Collagen_RNAseq.docx, P.SHO-250605-V1_Collagen_iPSC.docx
Data files: samples-250609-SC-R719C-CEL.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_Imaging.docx
Data files: TUNEL_quantification.xlsx, samples-250609-SC-R719C-CEL.xlsx, samples-250609-SC-R719C-D.IMG.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_RNAseq.docx
Data files: samples-250609-SC-R719C-CEL.xlsx, samples-250609-SC-R719C-RNA.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_RNAseq.docx
Data files: samples-250609-SC-R719C-DNA.xlsx, samples-250609-SC-R719C-RNA.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_RNAseq.docx
Data files: samples-250609-SC-R719C-D.SEQ.xlsx, samples-250609-SC-R719C-DNA.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_RNAseq.docx
Data files: samples-250609-SC-R719C-A.GEX.xlsx, samples-250609-SC-R719C-D.SEQ.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_Proteomics.docx
Data files: samples-250609-SC-R719C-CEL.xlsx, samples-250609-SC-R719C-D.MSP.xlsx
Snapshots: No snapshots
Submitter: Charles Demurjian
Assay type: Experimental Assay Type
Technology type: Technology Type
Investigation: Shoulders Lab Individual Publications
Organisms: No organisms
SOPs: P.SHO-250605-V1_Collagen_Proteomics.docx
Data files: samples-250609-SC-R719C-A.MSP.xlsx, samples-250609-SC-R719C-D.MSP.xlsx
Snapshots: No snapshots
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Studies: Human cartilage model of the precocious osteoar...
Assays: Cell Imaging - Data Linked
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Studies: Human cartilage model of the precocious osteoar...
Assays: Cell Imaging - Data Linked
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Studies: Human cartilage model of the precocious osteoar...
Assays: Library Creation - Metadata, RNA Extraction - Metadata
Batch sample publishing
Creator: Charles Demurjian
Submitter: Charles Demurjian
Creator: Charles Demurjian
Submitter: Charles Demurjian
Creator: Charles Demurjian
Submitter: Charles Demurjian
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications
Studies: Human cartilage model of the precocious osteoar...
Assays: Gene Expression Analysis - Data Linked, Library Creation - Metadata, RNA Extraction - Metadata, Short Read Sequencing - Data Linked, iPSC Culture and Chondronoid Formation - Metadata
Creator: Charles Demurjian
Submitter: Charles Demurjian
Investigations: Shoulders Lab Individual Publications