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Published on: August 2, 2022
Role of IFITM2 in osteogenic differentiation of C3H10T1/2 mesenchymal stem cells
Yongtao Zhang1, Xiangdong Li2, Shanshan Zhang1
1Key Laboratory for Biotech Drugs of the National Health Commission, Key Laboratory for Rare & Uncommon Diseases of Shandong Province, Biomedical Sciences College & Shandong Medicinal Biotechnology Centre, Shandong First Medical University & Shandong Academy of Medical Sciences, Ji'nan, Shandong, China.
Abstract:
Interferon-inducible transmembrane (IFITM) are a family of small proteins localized to plasma and endolysosomal membranes. Their functions beyond restricting viral entry and replication have been revealed in recent years. IFITM5 is involved in bone mineralization and is an osteogenic cell surface marker. IFITM1 and 3 interact with desmin and myosin, and are involved in myogenic differentiation. This study found upregulation of Ifitm2 during osteogenic differentiation of C3H10T1/2 cells. This positively correlated to the expression of osteogenic differentiation markers Col1a1, Alp, Runx2, and Ocn. Knockdown of Ifitm2 by siRNAs inhibited osteogenic differentiation, calcium deposition, and osteogenic marker expression of C3H10T1/2 cells. The osteoblast transcriptome revealed that knocking down Ifitm2 affected the expression Wnt signaling pathway-related genes, including Wnt family members, their receptors Lrp, Frizzled, and Lgr, and transmembrane molecule Rnf43 that suppresses the Wnt signaling pathway. Luciferase assays indicated enhancement of canonical Wnt signaling pathways by Ifitm2 overexpression. Furthermore, IFITM2 was colocalized in the metaphyseal bone and growth plate of the mouse tibial bone with SP7, a transcription factor essential for osteoblast differentiation and bone formation. These findings reveal a possible novel function and potential mechanisms of Ifitm2 in osteogenic differentiation.
Insights
Interferon-inducible transmembrane protein 2 (IFITM2) promotes osteogenic differentiation and bone formation by regulating the Wnt signaling pathway. Knocking down IFITM2 inhibits bone mineralization and osteogenic marker expression.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Interferon-inducible transmembrane (IFITM) proteins are known for roles in viral restriction.
- Recent studies reveal broader functions, including IFITM5 in bone mineralization and IFITM1/3 in muscle differentiation.
- The specific role of IFITM2 in osteogenesis was previously unclear.
Purpose of the Study:
- To investigate the role of Interferon-inducible transmembrane protein 2 (IFITM2) in osteogenic differentiation.
- To elucidate the molecular mechanisms underlying IFITM2's function in bone formation.
Main Methods:
- Upregulation analysis of Ifitm2 during osteogenic differentiation of C3H10T1/2 cells.
- siRNA-mediated knockdown of Ifitm2 to assess effects on differentiation and gene expression.
- Transcriptome analysis of osteoblasts with Ifitm2 knockdown.
- Luciferase assays to evaluate Wnt signaling pathway activity.
- Immunohistochemical colocalization studies in mouse bone tissue.
Main Results:
- Ifitm2 expression was upregulated during osteogenic differentiation, correlating with osteogenic markers (Col1a1, Alp, Runx2, Ocn).
- Ifitm2 knockdown inhibited osteogenic differentiation, calcium deposition, and expression of osteogenic markers.
- Knockdown of Ifitm2 altered the expression of Wnt signaling pathway components (Wnt ligands, receptors Lrp/Frizzled/Lgr, Rnf43).
- Ifitm2 overexpression enhanced canonical Wnt signaling.
- IFITM2 colocalized with the osteogenic transcription factor SP7 in mouse bone growth plates.
Conclusions:
- IFITM2 plays a novel and positive role in osteogenic differentiation.
- IFITM2 promotes bone formation, at least partly, through the regulation of the Wnt signaling pathway.
- IFITM2 is a potential therapeutic target for bone-related disorders.
