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Nature communications
01 13, 2021;12 (1): 362.

Single cell transcriptomic analysis of human pluripotent stem cell chondrogenesis.

Chia-Lung Wu, Amanda Dicks, Nancy Steward, Ruhang Tang, Dakota B Katz, Yun-Rak Choi, Farshid Guilak
Author Information
  1. Chia-Lung Wu: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  2. Amanda Dicks: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  3. Nancy Steward: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  4. Ruhang Tang: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  5. Dakota B Katz: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  6. Yun-Rak Choi: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA.
  7. Farshid Guilak: Dept. of Orthopaedic Surgery, Washington University in Saint Louis, St. Louis, MO, 63110, USA. guilak@wustl.edu. ORCID
PMID: 33441552 DOI: 10.1038/s41467-020-20598-y

Abstract

The therapeutic application of human induced pluripotent stem cells (hiPSCs) for cartilage regeneration is largely hindered by the low yield of chondrocytes accompanied by unpredictable and heterogeneous off-target differentiation of cells during chondrogenesis. Here, we combine bulk RNA sequencing, single cell RNA sequencing, and bioinformatic analyses, including weighted gene co-expression analysis (WGCNA), to investigate the gene regulatory networks regulating hiPSC differentiation under chondrogenic conditions. We identify specific WNTs and MITF as hub genes governing the generation of off-target differentiation into neural cells and melanocytes during hiPSC chondrogenesis. With heterocellular signaling models, we further show that WNT signaling produced by off-target cells is responsible for inducing chondrocyte hypertrophy. By targeting WNTs and MITF, we eliminate these cell lineages, significantly enhancing the yield and homogeneity of hiPSC-derived chondrocytes. Collectively, our findings identify the trajectories and molecular mechanisms governing cell fate decision in hiPSC chondrogenesis, as well as dynamic transcriptome profiles orchestrating chondrocyte proliferation and differentiation.

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Grants

  1. P30 AR073752/NIAMS NIH HHS
  2. R00 AR075899/NIAMS NIH HHS
  3. T32 DK108742/NIDDK NIH HHS
  4. R01 AG015768/NIA NIH HHS
  5. R21 AR067467/NIAMS NIH HHS
  6. K99 AR075899/NIAMS NIH HHS
  7. T32 EB018266/NIBIB NIH HHS
  8. R21 AR065956/NIAMS NIH HHS
  9. R01 AG046927/NIA NIH HHS
  10. P30 AR074992/NIAMS NIH HHS

MeSH Term

Animals
Benzeneacetamides
Cell Differentiation
Cell Line
Cells, Cultured
Chondrogenesis
Computational Biology
Gene Expression Profiling
Humans
Induced Pluripotent Stem Cells
Mice, Inbred NOD
Mice, Knockout
Mice, SCID
Pluripotent Stem Cells
Pyridines
Single-Cell Analysis
Transcriptome
Mice

Chemicals

2-(4-(2-methylpyridin-4-yl)phenyl)-N-(4-(pyridin-3-yl)phenyl)acetamide
Benzeneacetamides
Pyridines
Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.
Article has an altmetric score of 35

Word Cloud

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