Osteogenic growth peptide is a potent anti-inflammatory and bone preserving hormone via cannabinoid receptor type 2
- Department of Anatomy & Anthropology, Sackler Faculty of Medicine, Tel Aviv University, Israel
- Bone Laboratory, Institute of Dental Sciences, Faculty of Dental Medicine, Hebrew University of Jerusalem, Israel
- Institute for Drug Research, Faculty of Medicine, Hebrew University of Jerusalem, Israel
- Institute of Medical Sciences, University of Aberdeen, United Kingdom
- Department of Neurobiology, Weizmann Institute of Science, Israel
- Department of Oral and Maxillofacial Surgery, Hadassah-Hebrew University Hospital, Israel
- Institute of Molecular Psychiatry, University of Bonn, Germany
Figures
Figure 1 with 1 supplement
Aging differences in the endocannabinoid (EC) system.
(A) Literature scan isolated osteogenic growth peptide (OGP) as the only peptide that is mitogenic in osteoblasts and whose levels decline with age. (B–E) Age-related bone loss (decrease in …
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Figure 1—source data 1
BV/TV female and male mice 6 and 3 month old.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig1-data1-v2.xlsx
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Figure 1—source data 2
Cnr2 expression male and female 3 and 6 month mice.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig1-data2-v2.xlsx
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Figure 1—source data 3
Endocannabinoid levels in 3 and 6 month old mice.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig1-data3-v2.xlsx
Figure 1—figure supplement 1
Age-related differences in femoral bone.
Femurs were collected from female and male mice 6 month (6 mo) vs. 3-month-old (3 mo). Trabecular parameters were evaluated in the distal metaphysis; cortical parameters in the mid-diaphysis. (A) …
Figure 2 with 2 supplements
Osteogenic growth peptide (OGP) attenuates forskolin (FSK)-stimulated cAMP levels via cannabinoid receptor type 2 (CB2).
(A) CHO cells mock transfected with an empty vector (that does not express CB2) were treated with vehicle (Veh) or forskolin (FSK) to induce cAMP levels (with Veh only). FSK-treated cells were also …
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Figure 2—source data 1
Mock and hCB2-CHO cAMP levels (binding assay).
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig2-data1-v2.xlsx
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Figure 2—source data 2
cAMP levels in hCB2-CHO with CB2 antagonist (SR2).
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig2-data2-v2.xlsx
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Figure 2—source data 3
hCB1-HEK293 cAMP levels.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig2-data3-v2.xlsx
Figure 2—figure supplement 1
Osteogenic growth peptide (OGP) inhibits the forskolin-stimulated cAMP in CHO-hCB2 cells.
OGP and SR144528 (selective CB2 antagonist) were added at the indicated concentrations. The amount of [3H]cAMP in the absence of forskolin was subtracted. Data are the mean ± S.E.M. of two …
Figure 2—figure supplement 2
Osteogenic growth peptide (OGP) does not activate cannabinoid receptor type 1 (CB1) internal signaling in hCB1-HEK293 cells.
Forskolin (FSK) and the CB1 agonist HU210 increased cAMP levels in HEK293 cells transfected with human CNR1. Treatment with OGP had no effect on these CNR1-transfected cells. Data are mean ± SD …
Figure 3 with 1 supplement
Interaction of the osteogenic growth peptide (OGP) with the allosteric site of human cannabinoid receptor type 2 (CB2).
(A) and (B) represent conformation of the peptide when OGP is bound to the extracellular surface in the absence (A) and presence (B) of CP55940. The conformations presented are taken from the last …
Figure 3—figure supplement 1
Molecular dynamics (MD) energy image.
Total energy of the system during MD simulation of osteogenic growth peptide at the ECL site of the inactive model of human cannabinoid receptor type 2 (CB2) in the absence (A) and presence (B) of …
Figure 4 with 2 supplements
The effect of osteogenic growth peptide (OGP) in bone cells is dependent on cannabinoid receptor type 2 (CB2).
(A–B) Proliferative activity of HU910, a synthetic CB2 selective agonist (A), and of OGP (B) in wild type (WT) and Cnr2-/--derived murine osteoblasts. Data are the mean ± SD obtained in triplicate …
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Figure 4—source data 1
Murine WT and Cnr2-/- osteoblasts proliferation.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data1-v2.xlsx
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Figure 4—source data 2
Human osteoblasts proliferation with CB2 antagonist.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data2-v2.xlsx
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Figure 4—source data 3
Murine WT and Cnr2-/- osteoclasts differentiation.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data3-v2.xlsx
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Figure 4—source data 4
BrdU on WT osteoblasts with OGP.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data4-v2.xls
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Figure 4—source data 5
BrdU on Cnr2-/- osteoblasts with OGP.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data5-v2.xls
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Figure 4—source data 6
Murine osteoblasts with OGP, HU910 and CB2 antagonist.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig4-data6-v2.xlsx
Figure 4—figure supplement 1
The effect of osteogenic growth peptide (OGP) in murine bone cells is dependent on CB2.
Bromodeoxyuridin assay determination of the proliferative activity of OGP in wild type (WT) and Cnr2-/--derived new-born mouse calvarial osteoblasts. Data are the mean ± SD obtained in six …
Figure 4—figure supplement 2
The effect of osteogenic growth peptide (OGP) in murine bone cells is dependent on cannabinoid receptor type 2 (CB2).
(A,B) The proliferative activity of HU910 (A) and OGP (B) in murine osteoblasts is abrogated by the CB2 selective antagonist SR144528. Data are mean ± SD obtained in triplicates. *p<0.05 vs. vehicle …
Figure 5 with 1 supplement
Osteogenic growth peptide (OGP) effect on bone recovery in an ovariectomized (OVX) model is cannabinoid receptor type 2 dependent.
Rescue of wild type (WT) (A–F) but not Cnr2-/- (G–L) OVX-induced bone loss by OGP administration. 8-week old mice were OVX (or Sham-OVX), and treatment with OGP or vehicle started after 6 weeks for …
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Figure 5—source data 1
MicroCT morphometric data of OVX and Sham-OVX mice.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig5-data1-v2.xlsx
Figure 5—figure supplement 1
Osteogenic growth peptide (OGP) effect on bone recovery in ovariectomized (OVX) mice is cannabinoid receptor type 2 dependent.
Data from the same mice as in Figure 5. 8-week old mice were OVX (or Sham-OVX), and treatment with OGP or vehicle (Veh) started after 6 weeks for 6 weeks. (A, C) Cortical mid-diaphyseal diameter …
Figure 6
In vitro anti-inflammatory activity of osteogenic growth peptide (OGP) in wild type (WT) but not in Cnr2-/- macrophage cultures.
(A–B) Lipopolysaccharide (LPS)-induced expression of Tnfa (A) and Il1b (B) in WT macrophages pre-treated with OGP. (C–D) LPS-induced expression of Tnfa (C) and Il1b (D) in Cnr2-/- macrophages …
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Figure 6—source data 1
Murine WT and Cnr2 macrophages cytokine levels.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig6-data1-v2.xlsx
Figure 7
Osteogenic growth peptide (OGP) attenuates xylene-induced ear swelling.
(A) Wild type (WT) mice and (B) Cnr2-/- mice treated with PBS (vehicle [Veh]), indomethacin (positive control), HU910 (a selective CB2 agonist and positive control), or OGP prior to xylene …
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Figure 7—source data 1
Ear edema in WT and Cnr2-/- mice.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig7-data1-v2.xlsx
Figure 8 with 1 supplement
Osteogenic growth peptide (OGP[1-14]) levels in women and the effect of OGP on age-related bone loss in male mice.
(A) Immunoreactive (ir) OGP(1-14) levels were measured in serum from human female subjects aged 18–49 years. irOGP(1-14) levels were significantly lower in the fifth decade compared with the third …
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Figure 8—source data 1
OGP serum levels in women.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig8-data1-v2.xlsx
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Figure 8—source data 2
MicroCT data for 3- and 6-mo mice treated with OGP.
- https://cdn.elifesciences.org/articles/65834/elife-65834-fig8-data2-v2.xlsx
Figure 8—figure supplement 1
Effect of osteogenic growth peptide (OGP) on age-related cortical bone loss in male mice.
(A, B) Data are the mean ± SD for mid-diaphyseal diameter (A) and medullary diameter from the same mice as in Figure 8 showing a significant cortical expansion, n=8. *p<0.05 vs. vehicle …
Additional files
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Supplementary file 1
Literature search results summary, indicating the results of a Pubmed search (keywords [‘peptide’ or ‘protein’] and [‘Gi protein’ or ‘G(i)’ or ‘GPCR’ or ‘G*coupled receptor’] and [osteoblast* or osteoclast* OR osteocyte*]) where the activator interacts with a Gi-GPCR (26 agonists in total).
The 10 agonists that are also endogenous proteins/peptides are highlighted in bold fonts. PTX, pertussis toxin.
- https://cdn.elifesciences.org/articles/65834/elife-65834-supp1-v2.docx
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Supplementary file 2
Glide docking score (in kcal/mol) obtained for binding OGP in active and inactive models of CB2.
- https://cdn.elifesciences.org/articles/65834/elife-65834-supp2-v2.docx
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Supplementary file 3
Percent identity and similarity between CB1 and CB2 calculated using the online resource by Munk et al., 2019.
- https://cdn.elifesciences.org/articles/65834/elife-65834-supp3-v2.docx
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Transparent reporting form
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