Long non-coding RNA Malat1 fine-tunes bone homeostasis and repair by orchestrating cellular crosstalk and β-catenin-OPG/Jagged1 pathway
- Arthritis and Tissue Degeneration Program and David Z. Rosensweig Genomics Research Center, Hospital for Special Surgery, United States
- Department of Medicine, Weill Cornell Medical College, United States
- Research Institute, Hospital for Special Surgery, United States
- Department of Orthopaedic Surgery, Weill Cornell Medicine, United States
- Department of Cell and Developmental Biology, Cancer center at Illinois, University of Illinois at Urbana-Champaign, United States
- Institute for Tumor Biology and Experimental Therapy, Germany
- RNA Biology Laboratory, Faculty of Pharmaceutical Sciences, Hokkaido University, Japan
- Orthopaedic Trauma Service, Hospital for Special Surgery & NewYork-Presbyterian Hospital, United States
- Graduate Program in Cell and Development Biology, Weill Cornell Graduate School of Medical Sciences, United States
Figures
Figure 1 with 1 supplement
Malat1 deficiency disrupts bone remodeling and results in osteoporosis through reduced osteoblastic bone formation and increased osteoclastic bone resorption.
(A) μCT images and (B) bone morphometric analysis of trabecular bone of the distal femurs isolated from the 12-week-old-male (n=11, upper panel) and female (n=9, lower panel) WT and Malat1-/- …
-
Figure 1—source data 1
Malat1 deficiency disrupts bone remodeling and results in osteoporosis through reduced osteoblastic bone formation and increased osteoclastic bone resorption.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Malat1 deficiency does not affect mouse body weight and cortical bone.
(A) Body weight of 12-week-old male WT and Malat1-/- littermates. n=6 / group. (B) μCT images and bone morphometric analysis of cortical bone of the mid-shaft femurs isolated from 12-week-old male …
-
Figure 1—figure supplement 1—source data 1
Malat1 deficiency does not affect mouse body weight and cortical bone.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig1-figsupp1-data1-v1.xlsx
Figure 2 with 1 supplement
Specific deletion of Malat1 in osteoblasts leads to reduced bone mass and defects in bone formation.
(A) μCT images and (B) bone morphometric analysis of trabecular bone of the distal femurs isolated from the 12-week-old male Malat1f/f and Malat1 cKOOcn littermate mice (n = 14/group). (C) Images of …
-
Figure 2—source data 1
Specific deletion of Malat1 in osteoblasts leads to reduced bone mass and defects in bone formation.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Malat1 deficiency in Malat1ΔOcn mice does not affect body weight and cortical bone.
(A) Body weight of 12-week-old male Malat1f/f and Malat1 cKOOcn littermate mice. n=5/group. (B) μCT images and bone morphometric analysis of cortical bone of the mid-shaft femurs isolated from …
-
Figure 2—figure supplement 1—source data 1
Malat1 deficiency in Malat1 cKOOcn mice does not affect body weight and cortical bone.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig2-figsupp1-data1-v1.xlsx
Figure 3 with 2 supplements
Malat1 binds to β-catenin to positively regulate canonical Wnt/ β-catenin signaling pathway.
(A) ChIRP analysis of the specificity and efficiency of the Malat1 probe. Mouse Malat1 or the control GFP probes were used to pull down endogenous Malat1 from MC3T3-E1 cells, followed by qPCR …
-
Figure 3—source data 1
Malat1 binds to β-catenin to positively regulate canonical Wnt/ β-catenin signaling pathway.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig3-data1-v1.xlsx
-
Figure 3—source data 2
PDF file containing original western blots for Figure 3B and D, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig3-data2-v1.pdf
-
Figure 3—source data 3
Original files for western blot analysis displayed in Figure 3B and D.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig3-data3-v1.zip
Figure 3—figure supplement 1
Murine and human Malat1 probe sequences and their complementary Malat1 sequences.
Figure 3—figure supplement 2
Immunofluorescence staining of β-catenin.
Immunofluorescence staining of β-catenin (green) translocation into nuclei with or without Wnt3a treatment for 1 hr in calvarial osteoblasts isolated from the WT and Malat1-/- mice. Arrows: nuclear …
Figure 4
Malat1 is not an intrinsic regulator of osteoclast differentiation.
(A) Osteoclast differentiation using BMMs obtained from WT and Malat1-/- mice stimulated with RANKL for 3 days. TRAP staining (left panel) was performed and the area of TRAP-positive MNCs (≥3 …
-
Figure 4—source data 1
Malat1 is not an intrinsic regulator of osteoclast differentiation.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig4-data1-v1.xlsx
-
Figure 4—source data 2
PDF file containing original western blots for Figure 4D, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig4-data2-v1.pdf
-
Figure 4—source data 3
Original files for western blot analysis displayed in Figure 4D.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig4-data3-v1.zip
Figure 5
Malat1 promotes OPG expression in osteoblasts to suppress osteoclastogenesis.
(A) TRAP staining (left) and histomorphometric analysis (right) of histological sections obtained from the metaphysis region of distal femurs from the 12-week-old male Malat1f/f and Malat1 cKOOcn …
-
Figure 5—source data 1
Malat1 promotes OPG expression in osteoblasts to suppress osteoclastogenesis.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig5-data1-v1.xlsx
-
Figure 5—source data 2
PDF file containing original western blots for Figure 5H, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig5-data2-v1.pdf
-
Figure 5—source data 3
Original files for western blot analysis displayed in Figure 5H.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig5-data3-v1.zip
Figure 6 with 2 supplements
Malat1 enhances OPG and Jagged1 expression in chondrocytes.
(A) UMAP plot analysis of the bone and bone marrow datasets of scRNAseq based on GSE128423. (B) UMAP plot of the expression of Tnfrsf11b (encoding OPG) in bone and bone marrow cells. (C) Violin …
-
Figure 6—source data 1
Malat1 enhances OPG and Jagged1 expression in chondrocytes.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig6-data1-v1.xlsx
-
Figure 6—source data 2
PDF file containing original western blots for Figure 6G and I, indicating the relevant bands.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig6-data2-v1.pdf
-
Figure 6—source data 3
Original files for western blot analysis displayed in Figure 6G and I.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig6-data3-v1.zip
Figure 6—figure supplement 1
Bioinformatic analysis of the scRNAseq dataset GSE128423.
(A) A clustering tree of the scRNAseq dataset (GSE128423) across resolutions. (B) Dot plots of several typical marker gene expression for each cell type across the listed scRNAseq clusters. Cell …
Figure 6—figure supplement 2
Verification of the cellular characteristics of the primary chondrocytes isolated from mouse knees.
(A) Alcian blue staining of primary chondrocytes isolated from the WT and Malat1-/- littermate mice. (B) Immunofluorescence staining of aggrecan (green) of primary chondrocytes derived from WT and Ma…
Figure 7
A model illustrating a Malat1-centered molecular and cellular network in bone remodeling.
Malat1 binds to β-catenin, regulating its transcriptional activity on downstream target genes, such as Tnfrsf11b (encoding OPG) and Jag1 (encoding Jagged1), both of which are osteoclastogenic …
Figure 8
Malat1 enhances bone regeneration in fracture healing.
(A) Representative photograph of femur fracture callus. (B, D) representative μCT images of femurs isolated from the indicated mice at day 21 post-fracture. (C, E) μCT analysis of BV/TV in callus …
-
Figure 8—source data 1
Malat1 enhances bone regeneration in fracture healing.
- https://cdn.elifesciences.org/articles/98900/elife-98900-fig8-data1-v1.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (M. musculus) | Malat1-/- | PMID:22718948 | ||
| Genetic reagent (M. musculus) | Malat1flox/flox | PMID:22858678 | ||
| Genetic reagent (M. musculus) | Osteocalcin cre | PMID:12215457 | RRID:IMSR_JAX:019509 | |
| Genetic reagent (M. musculus) | Lyz2 cre | PMID:10621974 | RRID:IMSR_JAX:004781 | |
| Antibody | β-catenin rabbit polyclonal | Cell Signalling Technology | Cat# 9562 RRID:AB_331149 | WB (1:1000) |
| Antibody | Jag1 Rabbit monoclonal | Cell Signalling Technology | Cat# 70109 RRID:AB_331149 | WB (1:1000) |
| Antibody | Nfatc1 Mouse monoclonal | BD Biosciences | Cat# 556602 RRID:AB_331149 | WB (1:1000) |
| Antibody | Blimp1 Rat monoclonal | Santa Cruz Biotechnology | Cat# sc-47732 RRID:AB_628168 | WB (1:1000) |
| Antibody | c-Fos Rabbit polyclonal | Santa Cruz Biotechnology | Cat# sc-52 RRID:AB_2106783 | WB (1:1000) |
| Antibody | OPG/Osteoprotegerin Mouse monoclonal | Santa Cruz Biotechnology | Cat# sc-390518 RRID:AB_2891104 | WB (1:1000) |
| Antibody | p38α Rabbit polyclonal | Santa Cruz Biotechnology | Cat# sc-535 RRID:AB_632138 | WB (1:1000) |
| Antibody | Aggrecan Rabbit polyclonal | ABclonal | Cat# A8536 RRID:AB_632138 | IF (1:100) |
| Antibody | goat anti-Rabbit Alexa Fluor 488 | ThermoFisher Scientific | Cat# A-11008 RRID:AB_143165 | IF (1:500) |
| Antibody | ProLong Gold Antifade Mountant with DAPI | ThermoFisher Scientific | Cat# P36941 | |
| Peptide, recombinant protein | Recombinant Human sRANK Ligand | PeproTech | Cat# 310–01 | 40 ng/mL |
| Peptide, recombinant protein | Murine M-CSF | PeproTech | Cat# 315–02 | 20 ng/ml |
| Peptide, recombinant protein | Recombinant human OPG | PeproTech | Cat# 450–14 | Refer to Figure legends |
| Chemical compound, drug | prostaglandin E2 | MilliporeSigma | Cat# P0409 | 1 μM |
| Chemical compound, drug | 1α,25-Dihydroxyvitamin D3 | MilliporeSigma | Cat# D1530 | 10 nM |
| Chemical compound, drug | Collagenase | Worthington | Cat#LS004177 | 1 mg/ml |
| Chemical compound, drug | Dispase | Thermo Fisher Scientific | Cat#17105041 | 2 mg/ml |
| Commercial assay or kit | Mouse Tartrate Resistant Acid Phosphatase (TRAP) ELISA Kit | MyBioSource.com | MBS1601167 | |
| Commercial assay or kit | Mouse Osteoprotegerin ELISA Kit | MilliporeSigma | RAB0493 | |
| Commercial assay or kit | Mouse PINP ELISA Kit | MyBioSource.com | MBS2500076 | |
| Commercial assay or kit | Dual-Luciferase Reporter Assay system | Promega | E1910 | |
| Software, algorithm | Seurat | PMID:29608179 | RRID:SCR_016341 | https://satijalab.org/seurat/get_started.html |
| Software, algorithm | GraphPad Prism 8 | GraphPad Software | RRID:SCR_002798 | |
| Software, algorithm | ZEN (blue edition) version 3.4 | ZEN (blue edition) | RRID:SCR_013672 | https://www.zeiss.com/microscopy/en/products/software/zeiss-zen.html |
