GNAS/PKA signaling promotes aberrant osteochondral differentiation of Gli1+ tendon sheath progenitors

  • EMBO J. 2025 Sep 1. doi: 10.1038/s44318-025-00553-7.
Lijun Chen  #  1  2  3 ,  Chao Peng  #  2 ,  Lanyi Chai  #  2 ,  Renjie Zhang  #  1 ,  Chenghang Zhu  2 ,  Hailin Wang  2 ,  Qirong Cheng  2 ,  Yan Yan  4 ,  Cailiang Shen  1 ,  Hong Zheng  2 ,  Jiazhao Yang  #  5 ,  Haitao Fan  #  6 ,  Chen Kan  #  7  8  9
Affiliations
  • 1. Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, 230032, Hefei, China.
  • 2. Department of Pathophysiology, School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230022, Hefei, China.
  • 3. Department of Pathology, the Second People's Hospital of Hefei, Hefei Hospital Affiliated to Anhui Medical University, Guangde Street, 230011, Hefei, China.
  • 4. Laboratory Animal Research Center, School of Basic Medical Sciences, Anhui Medical University, He Fei, China.
  • 5. Department of Orthopedics, The First Affiliated Hospital of USTC, 17 Lujiang Road, 230001, Hefei, China. [email protected].
  • 6. Department of Orthopedics, The First Affiliated Hospital of Ningbo University, 59 Liuting Street, 315010, Ningbo, China. [email protected].
  • 7. Department of Orthopedics, The First Affiliated Hospital of Anhui Medical University, 218 Jixi Road, 230032, Hefei, China. [email protected].
  • 8. Department of Pathophysiology, School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230022, Hefei, China. [email protected].
  • 9. Laboratory Animal Research Center, School of Basic Medical Sciences, Anhui Medical University, He Fei, China. [email protected].
  • # Contributed equally.
PMID: 40890486 DOI: 10.1038/s44318-025-00553-7
Abstract

Tendon injury promotes aberrant osteochondral differentiation of tendon stem cells (TSCs) and results in disability. However, the cellular subsets within the osteochondral lineage involved in this process and associated mechanisms remain unclear. Here, we found that, following Achilles tenotomy, murine Gli1+ tendon sheath cells expanded rapidly, transitioning into tenogenic and osteochondrogenic cells. Lineage tracing, together with single-cell RNA Sequencing, revealed that osteochondrogenic Gli1+ tendon sheath cells originate from Scx+ tendon stem/progenitor cells, preferentially differentiate into osteochondral lineage tendon progenitors at 7 dpi, subsequently undergoing aberrant chondrogenesis and osteogenesis at 21dpi and 63dpi, respectively. In addition, Acvr1R206H/+ robustly accelerates osteochondral differentiation in Gli1+ tendon sheath progenitors. Furthermore, GNAS/PKA signaling was significantly activated in osteochondral differentiation of Gli1+ tendon sheath progenitors. Alternatively, treatment with the Gsα antagonist, NF449, or genetic inhibition of the PKA subunit, Prkaca, in Gli1+ sheath progenitors significantly alleviated aberrant osteochondral differentiation. NF449 also prevented osteochondral differentiation of human tendon stem cells. These findings identify Gli1+ tendon sheath progenitors with osteochondral differentiation capacity during heterotopic ossification via activation of GNAS/PKA signaling, suggesting PKA as a potentially effective therapeutic target to treat tendon ossification.

Keywords
Glioma-associated Oncogene Homolog 1 (Gli1); Guanine Nucleotide-Binding Protein (s) Alpha (Gnas); Heterotopic Ossification; Protein Kinase A (PKA); Tendon Sheath Progenitors.
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