Publications by authors named "Qinyu Ma"

12 Publications

  • Page 1 of 1

Osteoclast-derived small extracellular vesicles induce osteogenic differentiation via inhibiting ARHGAP1.

Mol Ther Nucleic Acids 2021 Mar 4;23:1191-1203. Epub 2021 Feb 4.

Department of Biomedical Materials Science, Third Military Medical University, Chongqing 400038, China.

Activated osteoclasts release large amounts of small extracellular vesicles (sEVs) during bone remodeling. However, little is known about whether osteoclast-derived sEVs affect surrounding cells. In this study, osteoclasts were generated by stimulating bone marrow macrophages (BMMs) with macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear actor κB ligand (RANKL). We performed microarray analysis of sEV-microRNAs (miRNAs)s secreted from osteoclast at different stages and identified four miRNAs that were highly expressed in mature osteoclast-derived sEVs. One of these miRNAs, miR-324, significantly induced osteogenic differentiation and mineralization of primary mesenchymal stem cells (MSCs) by targeting , a negative regulator of osteogenic differentiation. We next fabricated an sEV-modified scaffold by coating decalcified bone matrix (DBM) with osteoclast-derived sEVs, and the pro-osteogenic regeneration activities of the sEV-modified scaffold were validated in a mouse calvarial defect model. Notably, miR-324-enriched sEV-modified scaffold showed the highest capacity on bone regeneration, whereas inhibition of miR-324 in sEVs abrogated these effects. Taken together, our findings suggest that miR-324-contained sEVs released from mature osteoclast play an essential role in the regulation of osteogenic differentiation and potentially bridge the coupling between osteoclasts and MSCs.
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http://dx.doi.org/10.1016/j.omtn.2021.01.031DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7900016PMC
March 2021

Small extracellular vesicles deliver osteolytic effectors and mediate cancer-induced osteolysis in bone metastatic niche.

J Extracell Vesicles 2021 Feb 18;10(4):e12068. Epub 2021 Feb 18.

Department of Orthopedics Southwest Hospital Third Military Medical University Chongqing 400038 China.

Extracellular vesicles (EVs) play critical roles in regulating bone metastatic microenvironment through mediating intercellular crosstalks. However, little is known about the contribution of EVs derived from cancer cells to the vicious cycle of bone metastasis. Here, we report a direct regulatory mode between tumour cells and osteoclasts in metastatic niche of prostate cancer via vesicular miRNAs transfer. Combined analysis of miRNAs profiles both in tumour-derived small EVs (sEVs) and osteoclasts identified miR-152-3p as a potential osteolytic molecule. sEVs were enriched in miR-152-3p, which targets osteoclastogenic regulator MAFB. Blocking miR-152-3p in sEVs upregulated the expression of MAFB and impaired osteoclastogenesis in vitro. In vivo experiments of xenograft mouse model found that blocking of miR-152-3p in sEVs significantly slowed down the loss of trabecular architecture, while systemic inhibition of miR-152-3p using antagomir-152-3p reduced the osteolytic lesions of cortical bone while preserving basic trabecular architecture. Our findings suggest that miR-152-3p carried by prostate cancer-derived sEVs deliver osteolytic signals from tumour cells to osteoclasts, facilitating osteolytic progression in bone metastasis.
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http://dx.doi.org/10.1002/jev2.12068DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7892803PMC
February 2021

Osteoclast-derived apoptotic bodies couple bone resorption and formation in bone remodeling.

Bone Res 2021 Jan 11;9(1). Epub 2021 Jan 11.

Department of Orthopedics, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China.

Bone remodeling is precisely coordinated by bone resorption and formation. Apoptotic osteoclasts generate large amounts of apoptotic bodies (ABs) marking the end of the bone resorption phase, whereas the functions of osteoclast-derived ABs remain largely unknown. Here, we identified the molecular profile of ABs derived from osteoclasts at distinct differentiation stages and investigated their corresponding functions. ABs were isolated from apoptotic bone marrow macrophages, preosteoclasts, and mature osteoclasts induced by staurosporine. Proteomic signature analysis with liquid chromatography-tandem mass spectrometry suggested marked protein cargo differences among the different ABs. Further bioinformatic analysis showed that the proteomic signatures of the ABs were highly similar to those of their parental cells. Functionally, pOC-ABs induced endothelial progenitor cell differentiation and increased CD31Emcn endothelial cell formation in a murine bone defect model via their PDGF-BB cargo. mOC-ABs induced osteogenic differentiation of mesenchymal stem cells and facilitated osteogenesis via RANKL reverse signaling. In summary, we mapped the detailed proteomic landscapes of ABs derived from osteoclasts and showed that their potential biological roles are important in coupling bone formation with resorption during bone remodeling.
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http://dx.doi.org/10.1038/s41413-020-00121-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7801485PMC
January 2021

Osteoclast-derived apoptotic bodies show extended biological effects of parental cell in promoting bone defect healing.

Theranostics 2020 22;10(15):6825-6838. Epub 2020 May 22.

Department of Orthopedics, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.

Apoptotic bodies (ABs) traditionally considered as garbage bags that enclose residual components of dead cells are gaining increasing attentions due to their potential roles in intercellular communications. In bone turn over, at the end of bone resorption phase, most osteoclasts undergo apoptosis, generating large amounts of ABs. However, it remains unclear of the role of osteoclast-derived ABs in bone remodeling. : Staurosporine (STS) was used to apoptotic induction and differential centrifugation was used to isolate ABs. Western blotting, flowcytometry and Transmission electron microscopy (TEM) were performed for ABs identification, while whole transcriptome of ABs from osteoclasts at different stages was detected by RNA-seq. VENN analysis and gene set enrichment analysis (GSEA) were performed to compare the profile similarities between ABs and parental cells. efficacy of ABs on angiogenesis and osteogenesis were evaluated by tube formation assay and ALP staining. , calvarial defect mice model was used to assess the effects of ABs-modified decalcified bone matrix (DBM) scaffolds on angiogenesis and osteogenesis. : Here we mapped the whole transcriptome paralleled with small RNA profiling of osteoclast derived ABs at distinct differentiation stages. Whole transcriptome analysis revealed significant differences in RNA signatures among the ABs generated from osteoclasts at different stages. By comparing with parental osteoclast RNA profiles, we found that the transcriptome of ABs exhibited high similarities with the corresponding parental cells. Functionally, and studies showed that similar with the parental cells, pOC-ABs potentiated endothelial progenitor cell proliferation and differentiation, whereas mOC-ABs promoted osteogenic differentiation. The inherited biological effects of ABs were shown mediated by several enriched lncRNAs of which the interference abolished AB functions. : Our study revealed the total RNA profiles of osteoclast derived ABs and demonstrated their biological functions. Both gene set and functional analysis indicated that osteoclast derived ABs are biologically similar with the parental cells suggesting their bridging role in osteoclast-osteoblast coupling in bone remodeling.
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http://dx.doi.org/10.7150/thno.45170DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7295057PMC
May 2020

Non-coenzyme role of vitamin B1 in RANKL-induced osteoclastogenesis and ovariectomy induced osteoporosis.

J Cell Biochem 2020 07 26;121(7):3526-3536. Epub 2020 Feb 26.

Department of Orthopedics, Southwest Hospital, Third Military Medical University, Chongqing, China.

Vitamins B are co-enzymes participating in energy metabolic pathways. While some vitamins B are known affecting bone homeostasis, the effects of vitamin B1 (thiamine) on bone health remains unclear. In our study, we used cell counting kit-8, tartrate-resistant acid phosphatase stain, actin cytoskeleton stain, and pit formation assay to evaluate the effect of thiamine on osteoclast differentiation, formation, and function, respectively. Then we used dichloro-dihydro-fluorescein diacetate assay to investigate reactive oxygen species (ROS) generation and removal. Osteoporosis model by ovariectomy was established for animal experiments. We found that thiamine had inhibitory effect on osteoclast differentiation. And its inhibitory role on osteoclast differentiation is in a dose-dependent way. Mechanistically, ThDP suppresses intracellular ROS accumulation and unfolded protein response signaling during osteoclastogenesis via inhibiting Rac-Nox1/2/4 and intracellular inositol-requiring protein-1α/X-box-binding protein pathways, respectively. Osteoporotic mice treated with thiamine rich dietary showed better bone strength relative to thiamine deficient dietary. Our study explored the non-coenzyme inhibitory functions of B1 vitamin in receptor activator of nuclear factor κB ligand induced osteoclastogenesis and uncovered the significance of B1 vitamin in bone health.
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http://dx.doi.org/10.1002/jcb.29632DOI Listing
July 2020

Physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via regulating calcium signaling.

BMB Rep 2020 Mar;53(3):154-159

Department of Blood Purification, General Hospital of Shenyang Military Area Command, Shenyang 110000, China.

We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKLevoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulindependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway. [BMB Reports 2020; 53(3): 154-159].
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7118355PMC
March 2020

Vitamin B5 inhibit RANKL induced osteoclastogenesis and ovariectomy induced osteoporosis by scavenging ROS generation.

Am J Transl Res 2019 15;11(8):5008-5018. Epub 2019 Aug 15.

Department of Orthopedics, Southwest Hospital, Third Military Medical University (Army Medical University) Chongqing 400038, China.

B vitamins are a class of water-soluble vitamins that play important roles in cell metabolism. The participation of B vitamins in bone health has been recognized for decades. Pantothenic acid (vitamin B5) is mainly known for its wide variety of sources. However, the potential role of pantothenic acid in bone health and metabolism is still unclear. In this study, we found pantothenic acid has a dual effect on RANKL-induced osteoclastogenesis. Tartrate-resistant acid phosphatase (TRAP) stain shows that osteoclastogenesis was remarkably induced in a lower dosage of pantothenic acid (< 200 mM) and significantly inhibited while the pantothenic acid concentration increases to a certain extent (> 500 mM). We further confirmed this dual effect of pantothenic acid in osteoclastogenesis by detecting osteoclast formation and bone resorption using focal adhesion stain and pit formation, respectively. Mechanistically, we found phosphatidylinositol 3 kinase-protein kinase B (PI3K-Akt) pathway was activated in pre-osteoclasts (pOCs) after cultured with lower dosage of pantothenic acid; while the ROS generation was eliminated with upregulation of forkhead box O1 (FoxO1), forkhead box O2 (FoxO2) and NF-E2-related factor 2 (Nrf2) in pOCs after cultured with higher dosage of pantothenic acid. Finally, we used ovariectomized (OVX) mice to explore the potential role of pantothenic acid rich dietary in regulating bone metabolism , the result shows that pantothenic acid rich dietary can protect bone loss from estrogen deficiency. In brief, our study identified a new understanding of pantothenic acid in regulating osteoclastogenesis, revealed a therapeutic potential of pantothenic acid in prevention of bone loss related disorders.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6731427PMC
August 2019

Mature osteoclast-derived apoptotic bodies promote osteogenic differentiation via RANKL-mediated reverse signaling.

J Biol Chem 2019 07 5;294(29):11240-11247. Epub 2019 Jun 5.

Department of Orthopedics, Southwest Hospital, Third Military Medical University, Chongqing 400038, China

In bone remodeling, after a lifespan of ∼2 weeks, osteoclasts undergo apoptosis in each bone turnover cycle, resulting in generation of a large number of apoptotic bodies (ABs). However, the biological roles of osteoclast-derived ABs (OC-ABs) in bone remodeling have not been investigated and remain unknown. In this study, we stimulated bone marrow macrophages with receptor activator of NF-κB ligand (RANKL) to obtain both preosteoclasts and mature osteoclasts (mOCs). We then used alendronate to induce apoptosis in preosteoclasts and mOCs and generate the respective ABs and used flow cytometry and immunoblotting to characterize the sizes and immunogenic characteristics of the extracted ABs. We show that mOC-ABs are engulfed by preosteoblastic MC3T3-E1 cells and promote the viability of these cells. Among all osteoclast-derived extracellular vesicles, mOC-ABs had the highest osteogenic potency. We further observed that mOC-ABs had the highest vesicular receptor activator of NF-κB (RANK) levels among all types of osteoclast-derived extracellular vesicles. Of note, masking of vesicular RANK by soluble RANKL strongly abolished the osteogenic potency of osteoclast-derived ABs. Mechanistically, we found that mOC-ABs induce osteoblast differentiation by activatingPI3K/AKT/mechanistic target of rapamycin (mTOR)/ribosomal protein S6 kinase signaling. In conclusion, OC-ABs promote osteogenic differentiation by stimulating osteoblast differentiation via activation of RANKL reverse signaling. These findings provide important insights into the reversal phase between the bone resorption and formation stages during bone remodeling and identify an AB-dependent cellular signaling mechanism in osteoclast-osteoblast coupling.
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http://dx.doi.org/10.1074/jbc.RA119.007625DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6643026PMC
July 2019

IL-11 is essential in promoting osteolysis in breast cancer bone metastasis via RANKL-independent activation of osteoclastogenesis.

Cell Death Dis 2019 04 30;10(5):353. Epub 2019 Apr 30.

Department of Biomedical Materials Science, Third Military Medical University, Chongqing, 400038, China.

A variety of osteolytic factors have been identified from breast cancer cells leading to osteolysis, but less is known about which factor plays an essential role in the initiation process prior to the overt vicious osteolytic cycle. Here, we present in vitro and in vivo evidences to clarify the role of interleukin-11 (IL-11) as an essential contributor to breast cancer bone metastasis mediated osteolysis. Animal studies showed that bone specific metastatic BoM-1833 cells induce earlier onset of osteolysis and faster tumor growth compared with MCF7 and parental MDA-MB-231 cells in BALB/c-nu/nu nude mice. IL-11 was further screened and identified as the indispensable factor secreted by BoM-1833 cells inducing osteoclastogenesis independently of receptor activator of nuclear factor κB ligand (RANKL). Mechanistic investigation revealed that the JAK1/STAT3 signaling pathway as a downstream effector of IL-11, STAT3 activation further induces the expression of c-Myc, a necessary factor required for osteoclastogenesis. By inhibiting STAT3 phosphorylation, AG-490 was shown effective in reducing osteolysis and tumor growth in the metastatic niche. Overall, our results revealed the essential role and the underlying molecular mechanism of IL-11 in breast cancer bone metastasis mediated osteolysis. STAT3 targeting through AG-490 is a potential therapeutic strategy for mitigating osteolysis and tumor growth of bone metastatic breast cancer.
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http://dx.doi.org/10.1038/s41419-019-1594-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6491651PMC
April 2019

The role of physical forces in osteoclastogenesis.

J Cell Physiol 2019 08 8;234(8):12498-12507. Epub 2019 Jan 8.

Department of Biomedical Materials Science, Third Military Medical University, Chongqing, China.

The movements of life at every level from organs, tissues, cells to sub-cells, are all conducted in certain physical environments. In the human body, skeletal tissue among all connective tissues is influenced the most by physical forces. Studying the biological behavior of bone cells under different physical environments is helpful in further understanding bone homeostasis and metabolism. Among all bone cells, osteoclast (OC) and OC steered bone remodeling is one of the key points in bone metabolism. In the past few decades, people's understanding of OC was mostly limited to its involvement of bone resorption under physiological and pathological conditions. However, more and more studies started to focus on how physical forces affect the formation and differentiation of OC. This review tries to illustrate the knowledge up to date about how osteoclastogenesis is regulated by physical forces through direct and indirect ways, including fluid shear force, compressive force, and microgravity. The direct way describes the straightforward effects produced by different forces in osteoclastogenesis, whereas the indirect way describes the effects of different forces in osteoclastogenesis through regulation of other bone cells when a certain force is applied. Molecular mechanisms were analyzed and reviewed in both direct and indirect regulation by different forces. Finally, we discussed the status quo and tendency of related research, as well as other unresolved issues, and some future prospects.
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http://dx.doi.org/10.1002/jcp.28108DOI Listing
August 2019

Alendronate induces osteoclast precursor apoptosis via peroxisomal dysfunction mediated ER stress.

J Cell Physiol 2018 09 30;233(9):7415-7423. Epub 2018 Mar 30.

Department of Orthopedics, Southwest Hospital, Third Military Medical University (Army Medical University), Chongqing, China.

Nitrogen-containing bisphosphonates including alendronate (ALN) are the current first line antiresorptive drug in treating osteoporosis. In our study, we found that ALN administration impaired the secretion of platelet derived growth factor-BB (PDGF-BB), the most important angiogenic cytokines produced by preosteoclast (POC), in both sham and ovariectomized (OVX) mice. To further understand this phenomenon, we induced bone marrow macrophages (BMMs) to POCs in vitro and detected the effects of ALN particularly in POCs. The proapoptotic effect of ALN in POCs was confirmed by flow cytometry. On the molecular level, we found that farnesyl diphosphate synthase (FDPS) inhibition of ALN led to peroxisomal dysfunction and up regulation of cytoprotective protein glucose-regulated protein (GRP) 78. Peroxisomal dysfunction further induced endoplasmic reticulum (ER) stress in POCs and finally resulted in cell apoptosis marked by reduced expression of B-cell lymphoma 2 (Bcl-2) and increased expressions of CCAAT/enhancer binding protein homologous protein (CHOP), Bcl2 associated X (Bax), and cleaved caspase-3. We concluded that ALN has no selectivity in inhibiting POC and mature osteoclast. For POCs, ALN inhibition of FDPS leads to peroxisomal dysfunction, which further mediates ER stress and finally causes cell apoptosis. Considering that decreased angiogenesis is also an important issue in treating osteoporosis, how to preserve pro-angiogenic POCs while depleting mature osteoclasts is a problem worthy to be solved.
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http://dx.doi.org/10.1002/jcp.26587DOI Listing
September 2018

Estrogen Deficiency-Mediated M2 Macrophage Osteoclastogenesis Contributes to M1/M2 Ratio Alteration in Ovariectomized Osteoporotic Mice.

J Bone Miner Res 2018 05 23;33(5):899-908. Epub 2018 Mar 23.

Department of Biomedical Materials Science, School of Biomedical Engineering, Third Military Medical University, Chongqing, China.

In this study, for the first time we discovered that the M1/M2 macrophage phenotype ratio is increased in bone marrow of ovariectomized (OVX) osteoporotic C57BL/6 mice. Considering estrogen is the main variable, we assumed that estrogen participated in this alteration. To determine whether and how estrogen contributes to the change of the M1/M2 ratio, we first isolated bone marrow macrophages (BMMs) from mice femur and stimulated the cells with lipopolysaccharide (LPS)/interferon γ (IFN-γ) for M1 polarization and interleukin 4 (IL-4)/IL-13 for M2 polarization. M1 and M2 macrophages were then exposed to RANKL stimulation, we found that M2 macrophage but not M1 macrophage differentiated into functional osteoclast leading to increased M1/M2 ratio. Intriguingly, 17β-estradiol (E2) pretreatment prevented osteoclastogenesis from M2 macrophages. By constructing shRNA lentivirus interfering the expression of different estrogen receptors in M2 macrophages, we found that estrogen protects M2 macrophage from receptor activator of nuclear factor κB ligand (RANKL) stimulation selectively through estrogen receptor α (ERα) and the downstream blockage of NF-κB p65 nuclear translocation. Animal studies showed that ERα selective agonist 4,4',4″-(4-propyl-[1H]-pyrazole-1,3,5-triyl) trisphenol (PPT) was able to replicate the therapeutic effects of E2 in treating osteoporotic OVX mice. Together, our findings reveal that estrogen deficiency-mediated M2 macrophage osteoclastogenesis leads to increased M1/M2 ratio in OVX mice. Reducing the M1/M2 ratio is a potential therapeutic target in treating postmenopausal osteoporosis. © 2017 American Society for Bone and Mineral Research.
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http://dx.doi.org/10.1002/jbmr.3364DOI Listing
May 2018