Related Experiment Video
Updated: Aug 5, 2026
Real-Time Imaging of CCL5-Induced Migration of Periosteal Skeletal Stem Cells in Mice
Published on: September 16, 2020
NLRC5 Deficiency Delays Bone Healing by Inhibiting Osteogenic Differentiation of Bone Marrow-Derived Stem Cells and
Peiying Lyu1, Jianru Liu1, Yuanbo Wang1
1Department of Periodontology, Peking University School and Hospital of Stomatology, National Center for Stomatology, National Clinical Research Center for Oral Diseases, National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing 100081, China.
Abstract:
Modulating the immune microenvironment has become an emerging strategy for promoting functional bone regeneration, identifying key therapeutic targets remains challenging. Our previous work showed that Nucleotide-binding oligomerization domain-like receptor family caspase recruitment domain containing protein 5 (NLRC5) is involved in bone destruction associated with periodontitis, but its potential and mechanism in regulating bone tissue repair and regeneration have not been fully elucidated. A monocortical bone defect model was established in the mouse femur to assess the impact of NLRC5 on in situ bone healing and regeneration. Mouse bone marrow-derived mesenchymal stem cells (BMSCs) were isolated to evaluate the effects of NLRC5 on osteogenic differentiation, proliferation, and migration. RNA sequencing was used to explore the direct regulatory mechanism of NLRC5 on osteogenic differentiation of mouse BMSCs. Mass cytometry was employed to examine the effect of NLRC5 on the bone marrow immune microenvironment, followed by in vitro validation experiments. Loss of NLRC5 impaired the healing and regeneration of femoral bone defects in mice, and led to a high inflammatory state in the early stage of healing. The absence of NLRC5 inhibited the osteogenic differentiation ability of BMSCs, and could be restored by NLRC5 overexpression, which was achieved through activation of the phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT) signaling pathway. Mass cytometry data revealed that NLRC5 may serve as an important factor in maintaining the differentiation and maturation of regulatory T cells (Tregs). By modulating the levels of inflammatory cytokines, NLRC5 further influences the osteogenic differentiation of BMSCs. NLRC5 serves as a key regulator and promising candidate for bone repair. NLRC5 contributes to bone regeneration through a dual mechanism: it promotes BMSCs osteogenic differentiation, at least in part via the PI3K/AKT/β-catenin signaling pathway, and indirectly modulates the local immune microenvironment to facilitate bone repair.
Insights
Nucleotide-binding oligomerization domain-like receptor family caspase recruitment domain containing protein 5 (NLRC5) is crucial for bone regeneration. It promotes osteogenic differentiation and modulates the immune microenvironment, offering a promising therapeutic target for bone repair.
Area of Science:
- Immunology
- Regenerative Medicine
- Orthopedics
Background:
- Modulating the immune microenvironment is key for bone regeneration, but therapeutic targets are unclear.
- Nucleotide-binding oligomerization domain-like receptor family caspase recruitment domain containing protein 5 (NLRC5) was previously linked to bone destruction in periodontitis.
- The role of NLRC5 in bone tissue repair and regeneration remains largely unknown.
Purpose of the Study:
- To investigate the impact and mechanism of NLRC5 on bone healing and regeneration.
- To assess NLRC5's effect on mesenchymal stem cell osteogenic differentiation, proliferation, and migration.
- To explore NLRC5's role in regulating the bone marrow immune microenvironment.
Main Methods:
- Established a monocortical bone defect model in mouse femur.
- Isolated mouse bone marrow-derived mesenchymal stem cells (BMSCs) for in vitro studies.
- Utilized RNA sequencing and mass cytometry to analyze molecular mechanisms and immune cell populations.
Main Results:
- Loss of NLRC5 impaired femoral bone defect healing and increased early-stage inflammation.
- NLRC5 deficiency inhibited BMSC osteogenic differentiation, which was restored by NLRC5 overexpression via the PI3K/AKT pathway.
- NLRC5 influences regulatory T cell differentiation and modulates inflammatory cytokines, impacting BMSC osteogenesis.
Conclusions:
- NLRC5 is a critical regulator of bone regeneration, promoting BMSC osteogenic differentiation partly through the PI3K/AKT/β-catenin pathway.
- NLRC5 indirectly modulates the local immune microenvironment to enhance bone repair.
- NLRC5 represents a promising therapeutic candidate for bone repair strategies.