Adiponectin inhibits the differentiation and maturation of osteoclasts via the mTOR pathway in multiple myeloma - PubMed
Adiponectin inhibits the differentiation and maturation of osteoclasts via the mTOR pathway in multiple myeloma
Zhaoyun Liu et al. Int J Mol Med. 2020 Apr.
Abstract
The present study sought to investigate the correlation between adipose cytokines (visfatin, leptin and adiponectin) and markers of multiple myeloma bone disease, and to determine the effects and mechanism of action of adiponectin on the differentiation and maturation of osteoclasts in multiple myeloma (MM). The levels of visfatin, leptin and adiponectin were measured. Their association with the indices of myeloma tumor load and bone disease were analyzed. Reverse transcription‑quantitative PCR was used to detect the expression of receptor activator of nuclear factor‑κB ligand (RANKL), osteoclast associated Ig‑like receptor (OSCAR), tartrate‑resistant acid phosphatase (TRAP) and Cathepsin K genes. Flow cytometry was used to detect the expression of adiponectin receptor 1 (AdipoR1) and the phosphorylation of the mechanistic target of rapamycin kinase (mTOR) pathway‑associated proteins mTOR and eukaryotic translation initiation factor 4E‑binding protein (4EBP1). There were no significant correlations among leptin, visfatin and the indexes of myeloma tumor load and bone disease. Serum adiponectin levels were significantly lower in patients with newly diagnosed multiple myeloma compared with healthy volunteers (12.37±3.13 vs. 13.80±0.95; P<0.05). The number of mature osteoclasts in the adiponectin group was lower compared with in the control group. Adiponectin also inhibited the mRNA expression of the osteoclast‑associated factors RANKL, OSCAR, TRAP and Cathepsin K. Comparison between the non‑adiponectin group and the adiponectin group revealed that adiponectin increased the expression of AdipoR1 on the surface of osteoclast precursor cells (26.21±4.27% vs. 29.86±6.23%; P<0.05) and reduced the expression of phosphorylated (p‑)mTOR (7.89±1.00% vs. 5.91±1.26%; P<0.05) and p‑4EBP1 (26.78±5.00% vs. 22.49±4.24%; P<0.05). The p‑mTOR and p‑4EBP1 levels in the adiponectin + MHY1485 (an mTOR signaling pathway‑specific agonist) group were significantly higher compared with those in the adiponectin group. It was revealed that adiponectin may inhibit osteoclast differentiation and maturation via the mTOR pathway. In conclusion, adiponectin inhibits the differentiation and maturation of osteoclasts by increasing the expression of AdipoR1 and reducing the phosphorylation levels of mTOR and 4EBP1 in patients with MM.
Figures
![Figure 1](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/c4ad57145b70/IJMM-45-04-1112-g00.gif)
Visfatin, leptin, and adiponectin levels in newly diagnosed patients with MM. *P<0.05 with comparisons shown by lines. MM, multiple myeloma.
![Figure 2](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/b0b511a6d504/IJMM-45-04-1112-g01.gif)
Correlation between adiponectin, and either OCN or CTX levels in patients with multiple myeloma. (A) Serum level of adiponectin is positively correlated with OCN levels. (B) Serum level of adiponectin is negatively correlated with CTX levels. OCN, osteocalcin; CTX, carboxy-terminal cross-linking telopeptide of type I collagen.
![Figure 3](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/e3c1b5e49ac3/IJMM-45-04-1112-g02.gif)
Adiponectin inhibits osteoclast differentiation and maturation. TRAP staining was performed in (A) the control group and (B) the adipo-nectin-treated group (original magnification, ×100). (C) Quantified number of TRAP-positive multinucleated cells. *P<0.05 with comparisons shown by lines. TRAP, tartrate-resistant acid phosphatase.
![Figure 4](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/5ae04ece5276/IJMM-45-04-1112-g03.gif)
mRNA levels of the osteoclast-specific factors RANKL, OSCAR, TRAP and Cathepsin K. Expression levels of mRNA are presented for the control and adiponectin-treated group for (A) RANKL, (B) OSCAR, (C) TRAP and (D) Cathepsin K. *P<0.05 with comparisons shown by lines. RANKL, receptor activator of nuclear factor-κB ligand; OSCAR, osteoclast associated Ig-like receptor; TRAP, tartrate-resistant acid phosphatase.
![Figure 5](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/373e5803d754/IJMM-45-04-1112-g04.gif)
Adiponectin upregulates the AdipoR1 expression level. AdipoR1 levels in (A) the control group and (B) the adiponectin-treated group. (C) Quantification of the AdipoR1 levels in the two groups (26.21±4.27 vs. 29.86±6.23%). *P<0.05 with comparisons shown by lines. AdipoR1, adiponectin receptor 1.
![Figure 6](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/935f/7053860/890199fcf618/IJMM-45-04-1112-g05.gif)
Phosphorylation levels of the mTOR pathway. (A) Osteoclast precursor was revealed to be CD14-positive. (B) Isotype control of p-mTOR and p-4EBP1 marked with IgG1-PE. (C) p-mTOR levels in the control group, adiponectin-treated group and adiponectin + MHY1485 group. (D) Histogram of the phosphorylation levels of the mTOR pathway. (E) p-4EBP1 levels in the control group, adiponectin-treated group and adiponectin + MHY1485 group. *P<0.05 with comparisons shown by lines. mTOR, mechanistic target of rapamycin kinase; CD14, cluster of differentiation 14; p-, phosphorylated; 4EBP1, eukaryotic translation initiation factor 4E-binding protein; IgG1, immunoglobulin G1; PE, phycoerythrin.
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