1. Academic Validation
  2. Differentiation of Hypertrophic Chondrocytes from Human iPSCs for the In Vitro Modeling of Chondrodysplasias

Differentiation of Hypertrophic Chondrocytes from Human iPSCs for the In Vitro Modeling of Chondrodysplasias

  • Stem Cell Reports. 2021 Mar 9;16(3):610-625. doi: 10.1016/j.stemcr.2021.01.014.
Yann Pretemer 1 Shunsuke Kawai 2 Sanae Nagata 1 Megumi Nishio 3 Makoto Watanabe 4 Sakura Tamaki 5 Cantas Alev 6 Yoshihiro Yamanaka 6 Jing-Yi Xue 7 Zheng Wang 8 Kenichi Fukiage 9 Masako Tsukanaka 10 Tohru Futami 10 Shiro Ikegawa 7 Junya Toguchida 11
Affiliations

Affiliations

  • 1 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan.
  • 2 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Department of Regeneration Science and Engineering, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan; Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
  • 3 Department of Regeneration Science and Engineering, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.
  • 4 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Life Science Research Center, Technology Research Laboratory, Shimadzu Corporation, Kyoto, Japan.
  • 5 Department of Regeneration Science and Engineering, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan; Institute for Advancement of Clinical and Translational Sciences, Kyoto University Hospital, Kyoto University, Kyoto, Japan.
  • 6 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Institute for the Advanced Study of Human Biology, Kyoto University, Kyoto, Japan.
  • 7 Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Tokyo, Japan.
  • 8 Laboratory for Bone and Joint Diseases, RIKEN Center for Integrative Medical Sciences, Tokyo, Japan; McKusick-Zhang Center for Genetic Medicine and State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, China.
  • 9 Department of Pediatric Orthopaedics, Shiga Medical Center for Children, Moriyama, Japan; Department of Orthopaedic Surgery, Bobath Memorial Hospital, Osaka, Japan.
  • 10 Department of Pediatric Orthopaedics, Shiga Medical Center for Children, Moriyama, Japan.
  • 11 Department of Cell Growth and Differentiation, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan; Department of Regeneration Science and Engineering, Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan; Department of Orthopaedic Surgery, Graduate School of Medicine, Kyoto University, Kyoto, Japan; Institute for Advancement of Clinical and Translational Sciences, Kyoto University Hospital, Kyoto University, Kyoto, Japan. Electronic address: [email protected].
Abstract

Chondrodysplasias are hereditary diseases caused by mutations in the components of growth cartilage. Although the unfolded protein response (UPR) has been identified as a key disease mechanism in mouse models, no suitable in vitro system has been reported to analyze the pathology in humans. Here, we developed a three-dimensional culture protocol to differentiate hypertrophic chondrocytes from induced pluripotent stem cells (iPSCs) and examine the phenotype caused by MATN3 and COL10A1 mutations. Intracellular MATN3 or COL10 retention resulted in increased ER stress markers and ER size in most mutants, but activation of the UPR was dependent on the mutation. Transcriptome analysis confirmed a UPR with wide-ranging changes in bone homeostasis, extracellular matrix composition, and lipid metabolism in the MATN3 T120M mutant, which further showed altered cellular morphology in iPSC-derived growth-plate-like structures in vivo. We then applied our in vitro model to drug testing, whereby trimethylamine N-oxide led to a reduction of ER stress and intracellular MATN3.

Keywords

COL10A1; MATN3; chondrodysplasia; iPSC; unfolded protein response.

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