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Suppression of TLR4 prevents diabetic bone loss by regulating FTO-mediated m6A modification
Ximei Shen1, Chao Lan1, Youfen Lin1
1Department of Endocrinology, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China; Department of Endocrinology, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou 350212, China; Clinical Research Center for Metabolic Diseases of Fujian Province, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China; Fujian Key Laboratory of Glycolipid and Bone Mineral Metabolism, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China; Diabetes Research Institute of Fujian Province, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China; Metabolic Diseases Research Institute, The First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China.
Abstract:
Toll-like receptor-4 (TLR4) has been implicated in the development and progression of diabetic osteoporosis. However, the mechanisms underlying TLR4-regulated bone metabolism in diabetes are yet to be fully understood. Epigenetic modifications have been indicated as a possible mechanism leading to increased risk of osteoporosis and bone fracture. As N6-methyladenosine (m6A) is the most common epigenetic modification in eukaryotic mRNAs, we hypothesized that TLR4 regulates m6A modification in bone tissues of diabetic rats, thereby potentially explaining the pathogenesis of diabetic bone loss. m6A sequencing (m6A-seq) was performed in samples of the femur of TLR4-wild type (TLR4WT) and TLR4-knockout (TLR4KO) diabetic rats to identify genes with differential m6A modifications that may be associated with the bone loss phenotype. We found that in TLR4KO rats, the rapid weight loss of diabetic rats was prevented, and bone mineral density (BMD) was significantly increased. m6A-seq and Gene Ontology enrichment analysis revealed that m6A-modified genes in the femur of TLR4KO diabetic rats were associated with regulation of biological processes such as osteoclast differentiation. qRT-PCR analysis on the expression levels of the m6A-modified methyltransferases and demethylases demonstrated that only the m6A demethylase fat mass and obesity-associated protein（FTO）was decreased. Using an osteoclast cell model, we confirmed that TLR4-mediated osteoclast differentiation was induced by glycolipid toxicity via inhibition of FTO expression. Taken together, these results suggest that inhibition of TLR4 may prevent diabetic bone loss via regulation of FTO-mediated m6A modification.
Insights
Toll-like receptor-4 (TLR4) inhibition prevents diabetic bone loss by regulating N6-methyladenosine (m6A) modification. This epigenetic change impacts osteoclast differentiation and bone mineral density in diabetic rats.
Area of Science:
- Molecular Biology
- Endocrinology
- Bone Metabolism
Background:
- Diabetic osteoporosis is a growing concern, with Toll-like receptor-4 (TLR4) implicated in its development.
- The precise mechanisms of TLR4's role in diabetic bone metabolism and the contribution of epigenetic modifications remain unclear.
- N6-methyladenosine (m6A) is a prevalent mRNA epigenetic modification potentially linked to osteoporosis risk.
Purpose of the Study:
- To investigate whether TLR4 regulates m6A modification in the bone tissue of diabetic rats.
- To elucidate the role of TLR4-mediated m6A changes in the pathogenesis of diabetic bone loss.
Main Methods:
- N6-methyladenosine sequencing (m6A-seq) was performed on femur samples from wild-type and TLR4-knockout diabetic rats.
- Gene Ontology enrichment analysis was used to identify differentially modified genes and associated biological processes.
- Quantitative reverse transcription PCR (qRT-PCR) assessed the expression of m6A methyltransferases and demethylases.
- An in vitro osteoclast cell model was utilized to confirm TLR4-mediated effects.
Main Results:
- TLR4 knockout in diabetic rats prevented weight loss and significantly increased bone mineral density (BMD).
- m6A-seq identified differential m6A modifications in genes associated with osteoclast differentiation in TLR4-knockout rats.
- The m6A demethylase, fat mass and obesity-associated protein (FTO), showed decreased expression in TLR4-knockout diabetic rats.
- TLR4-mediated osteoclast differentiation was confirmed to be induced by glycolipid toxicity through FTO inhibition.
Conclusions:
- TLR4 plays a critical role in regulating m6A modification within bone tissue during diabetes.
- Inhibition of TLR4 may serve as a therapeutic strategy to prevent diabetic bone loss.
- The FTO-mediated m6A pathway is a key mechanism through which TLR4 influences diabetic bone metabolism.
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