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Specific RNA m6A modification sites in bone marrow mesenchymal stem cells from the jawbone marrow of type 2 diabetes patients with dental implant failure

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International Journal of Oral Science 2023年第1期15卷 50-62页

作者： Wanhao Yan Xiao Lin Yiqian Ying Jun Li Zhipeng Fan Laboratory of Molecular Signaling and Stem Cells Therapy Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionCapital Medical University School of StomatologyBeijingChina Department of Dental Implant Center Beijing Stomatological HospitalSchool of StomatologyCapital Medical UniversityBeijingChina Department of Oral Implantology Tianjin Stomatological HospitalSchool of MedicineNankai UniversityTianjinChina Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijingChina

The failure rate of dental implantation in patients with well-controlled type 2 diabetes mellitus(T2DM)is higher than that in nondiabetic patients.This due,in part,to the impaired function of bone marrow mesenchymal stem cells(BMSCs)from the jawbone marrow of T2DM patients(DM-BMSCs),limiting implant osseointegration.RNA N6-methyladenine(m6A)is important for BMSC function and diabetes regulation.However,it remains unclear how to best regulate m6A modifications in DM-BMSCs to enhance function.Based on the“m6A site methylation stoichiometry”of m6A single nucleotide arrays,we identified 834 differential m6Amethylated genes in DM-BMSCs compared with normal-BMSCs(N-BMSCs),including 43 and 790 m6A hypermethylated and hypomethylated genes,respectively,and 1 gene containing hyper-and hypomethylated m6A sites.Differential m6A hypermethylated sites were primarily distributed in the coding sequence,while hypomethylated sites were mainly in the 3’-untranslated region.The largest and smallest proportions of m6A-methylated genes were on chromosome 1 and 21,respectively.Maz F-PCR and real-time RTPCR results for the validation of erythrocyte membrane protein band 4.1 like 3,activity-dependent neuroprotector homeobox(ADNP),growth differentiation factor 11(GDF11),and regulator of G protein signalling 2 agree with m6A single nucleotide array results;ADNP and GDF11 m RNA expression decreased in DM-BMSCs.Furthermore,gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses suggested that most of these genes were enriched in metabolic processes.This study reveals the differential m6A sites of DM-BMSCs compared with N-BMSCs and identifies candidate target genes to enhance BMSC function and improve implantation success in T2DM patients.

关键词： patients impaired implantation

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Insulin-like growth factor binding proteins 7 prevents dental pulp-derived mesenchymal stem cell senescence via metabolic downregulation of p21

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Science China(Life Sciences) 2022年第11期65卷 2218-2232页

作者： Xiaoyu Li Liang Feng Chunmei Zhang Jinsong Wang Songlin Wang Lei Hu Molecular Laboratory for Gene Therapy and tooth regeneration Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionBeijing Laboratory of Oral HealthCapital Medical University School of StomatologyBeijing 100050China Immunology research Center for Oral and Systemic Health Beijing Friendship HospitalCapital Medical UniversityBeijing 100050China Department of Biochemistry and Molecular Biology Capital Medical University School of Basic MedicineBeijing 100069China Laboratory for Oral and General Health Integration and Translation Beijing Tiantan HospitalCapital Medical UniversityBeijing 100070China Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijing 100700China Department of Prosthodontics Capital Medical University School of StomatologyBeijing 100050China

Cellular senescence affects the efficacy of mesenchymal stem cells(MSCs)-mediated tissue regeneration.Insulin-like growth factor binding proteins-7(IGFBP7),as a member of the IGF family,is associated with osteogenic differentiation and the senescence of MSCs,but its exact function and mechanism remain unclear.We found IGFBP7 promoted the osteogenic differentiation and prevented the senescence of dental pulp-derived MSCs(DPSCs),as observed in the gain-of-function and lossof-function analyses,the senescence-associated marker p21 showed the most pronounced expression changes.We demonstrated that IGFBP7 activated the biological activity of SIRT1 deacetylase via metabolism,resulting in a deacetylation of H3K36ac and a decrease of the binding affinity of H3K36ac to p21 promoter,thereby reducing the transcription of p21,which ultimately prevents DPSCs senescence and promotes tissue regeneration.The activation of the mitochondrial electron transport chain(ETC)by Coenzyme Q10 could rescue the promotion of DPSC senescence induced by the knockdown of IGFBP7,whereas the inhibition of ETC by rotenone attenuated the prevention of DPSC senescence induced by IGFBP7 overexpression.In conclusion,our present results reveal a novel function of IGFBP7 in preventing DPSC senescence via the metabolism-induced deacetylation of H3K36ac and reduction of p21 transcription,suggesting that IGFBP7 is a potential target for promoting tissue regeneration in an aging environment.

关键词： dental pulp-derived mesenchymal stem cells insulin-like growth factor binding protein 7 senescence metabolism SIRT1 p21

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MLL1 inhibits the neurogenic potential of SCAPs by interacting with WDR5 and repressing HES1

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International Journal of Oral Science 2023年第4期15卷 649-663页

作者： Chen Zhang Weilong Ye Mengyao Zhao Lujue Long Dengsheng Xia Zhipeng Fan Laboratory of Molecular Signaling and Stem Cells Therapy Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionBeijing Stomatological HospitalSchool of StomatologyCapital Medical UniversityBeijingChina Department of Dental Emergency Beijing Stomatological HospitalSchool of StomatologyCapital Medical UniversityBeijingChina Beijing Laboratory of Oral Health Capital Medical UniversityBeijingChina Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijingChina

Mesenchymal stem cell(MSC)-based therapy has emerged as a promising treatment for spinal cord injury(SCI),but improving the neurogenic potential of MSCs remains a challenge.Mixed lineage leukemia 1(MLL1),an H3K4me3 methyltransferases,plays a critical role in regulating lineage-specific gene expression and influences neurogenesis.In this study,we investigated the role and mechanism of MLL1 in the neurogenesis of stem cells from apical papilla(SCAPs).We examined the expression of neural markers,and the nerve repair and regeneration ability of SCAPs using dynamic changes in neuron-like cells,immunofluorescence staining,and a SCI model.We employed a coimmunoprecipitation(Co-IP)assay,real-time RT-PCR,microarray analysis,and chromatin immunoprecipitation(ChIP)assay to investigate the molecular mechanism.The results showed that MLL1 knock-down increased the expression of neural markers,including neurogenic differentiation factor(NeuroD),neural cell adhesion molecule(NCAM),tyrosine hydroxylase(TH),βIII-tubulin and Nestin,and promoted neuron-like cell formation in SCAPs.In vivo,a transplantation experiment showed that depletion of MLL 1 in SCAPs can restore motor function in a rat SCI model.MLL1 can combine with WD repeat domain 5(WDR5)and WDR5 inhibit the expression of neural markers in SCAPs.MLL1 regulates Hairy and enhancer of split 1(HES1)expression by directly binds to HES1 promoters via regulating H3K4me3 methylation by interacting with WDR5.Additionally,HES1 enhances the expression of neural markers in SCAPs.Our findings demonstrate that MLL1 inhibits the neurogenic potential of SCAPs by interacting with WDR5 and repressing HES1.These results provide a potential therapeutic target for promoting the recovery of motor function in SCI patients.

关键词： HES1 neurogenic MLL1

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miR-140-3p enhanced the osteo/odontogenic differentiation of DPSCs via inhibiting KMT5B under hypoxia condition

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International Journal of Oral Science 2021年第4期13卷 398-407页

作者： Han Zheng Ning Wang Le Li Lihua Ge Haichao Jia Zhipeng Fan Laboratory of Molecular Signaling and Stem Cells Therapy Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionCapital Medical University School of StomatologyBeijingChina Tsinghua University Hospital Stomatological Disease Prevention and Control CenterTsinghua UniversityBeijingChina Department of Orthodontics Capital Medical University School of StomatologyBeijingChina Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijingChina

Human dental pulp stem cells(DPSCs)have emerged as an important source of stem cells in the tissue engineering,and hypoxia will change various innate characteristics of DPSCs and then affect dental tissue regeneration.Nevertheless,little is known about the complicated molecular mechanisms.In this study,we aimed to investigate the influence and mechanism of miR-140-3p on DPSCs under hypoxia condition.Hypoxia was induced in DPSCs by Cobalt chloride(CoCl_(2))treatment.The osteo/dentinogenic differentiation capacity of DPSCs was assessed by alkaline phosphatase(ALP)activity,Alizarin Red S staining and main osteo/dentinogenic markers.A luciferase reporter gene assay was performed to verify the downstream target gene of miR-140-3p.This research exhibited that miR-140-3p promoted osteo/dentinogenic differentiation of DPSCs under normoxia environment.Furthermore,miR-140-3p rescued the CoCl_(2)-induced decreased osteo/odontogenic differentiation potentials in DPSCs.Besides,we investigated that miR-140-3p directly targeted lysine methyltransferase 5B(KMT5B).Surprisingly,we found inhibition of KMT5B obviously enhanced osteo/dentinogenic differentiation of DPSCs both under normoxia and hypoxia conditions.In conclusion,our study revealed the role and mechanism of miR-140-3p for regulating osteo/dentinogenic differentiation of DPSCs under hypoxia,and discovered that miR-140-3p and KMT5B might be important targets for DPSC-mediated tooth or bone tissue regeneration.

关键词： DPSCs miR inhibiting

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Molecular mechanisms of cellular metabolic homeostasis in stem cells

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International Journal of Oral Science 2023年第4期15卷 527-539页

作者： Xiaoyu Li Ou Jiang Songlin Wang Salivary Gland Disease Center and Beijing Key Laboratory of tooth regeneration and Function Reconstruction Beijing Laboratory of Oral Health and Beijing Stomatological HospitalCapital Medical UniversityBeijingChina Department of Biochemistry and Molecular Biology School of Basic Medical SciencesCapital Medical UniversityBeijingChina Immunology research Center for Oral and Systemic Health Beijing Friendship HospitalCapital Medical UniversityBeijingChina Laboratory for Oral and General Health Integration and Translation Beijing Tiantan HospitalCapital Medical UniversityBeijingChina Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijingChina

Many tissues and organ systems have intrinsic regeneration capabilities that are largely driven and maintained by tissue-resident stem cell populations. In recent years, growing evidence has demonstrated that cellular metabolic homeostasis plays a central role in mediating stem cell fate, tissue regeneration, and homeostasis. Thus, a thorough understanding of the mechanisms that regulate metabolic homeostasis in stem cells may contribute to our knowledge on how tissue homeostasis is maintained and provide novel insights for disease management. In this review, we summarize the known relationship between the regulation of metabolic homeostasis and molecular pathways in stem cells. We also discuss potential targets of metabolic homeostasis in disease therapy and describe the current limitations and future directions in the development of these novel therapeutic targets.

关键词： homeostasis maintained mechanisms

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Primary cilia in hard tissue development and diseases

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Frontiers of Medicine 2021年第5期15卷 657-678页

作者： Sijin Li Han Zhang Yao Sun Department of Orthodontics School&Hospital of StomatologyTongji UniversityShanghai Engineering Research Center of Tooth Restoration and RegenerationShanghai 200072China Department of Implantology School&Hospital of StomatologyTongji UniversityShanghai Engineering Research Center of Tooth Restoration and RegenerationShanghai 200072China

Bone and teeth are hard tissues.Hard tissue diseases have a serious effect on human survival and quality of life.Primary cilia are protrusions on the surfaces of cells.As antennas,they are distributed on the membrane surfaces of almost all mammalian cell types and participate in the development of organs and the maintenance of homeostasis.Mutations in cilium-related genes result in a variety of developmental and even lethal diseases.Patients with multiple ciliary gene mutations present overt changes in the skeletal system,suggesting that primary cilia are involved in hard tissue development and reconstruction.Furthermore,primary cilia act as sensors of external stimuli and regulate bone homeostasis.Specifically,substances are trafficked through primary cilia by intraflagellar transport,which affects key signaling pathways during hard tissue development.In this review,we summarize the roles of primary cilia in long bone development and remodeling from two perspectives:primary cilia signaling and sensory mechanisms.In addition,the cilium-related diseases of hard tissue and the manifestations of mutant cilia in the skeleton and teeth are described.We believe that all the findings will help with the intervention and treatment of related hard tissue genetic diseases.

关键词： primary cilia bone mechanical sensing hard tissue cilium-related bone disease tooth

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IFT140^(+)/K14^(+) cells function as stem/progenitor cells in salivary glands

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International Journal of Oral Science 2022年第4期14卷 561-571页

作者： Xueming Zhang Ji Zhou Xinyu Wang Jiangyu Geng Yubei Chen Yao Sun Department of Oral and Maxillofacial Surgery Stomatological Hospital and Dental School of Tongji UniversityShanghai Engineering Research Center of Tooth Restoration and RegenerationNo.399YanChang Middle RoadShanghaiChina Department of Implantology Stomatological Hospital and Dental School of Tongji UniversityShanghai Engineering Research Center of Tooth Restoration and RegenerationNo.399YanChang Middle RoadShanghaiChina

Stem/progenitor cells are important for salivary gland development, homeostasis maintenance, and regeneration following injury.Keratin-14^(+)(K14^(+)) cells have been recognized as bona fide salivary gland stem/progenitor cells. However, K14 is also expressed in terminally differentiated myoepithelial cells;therefore, more accurate molecular markers for identifying salivary stem/progenitor cells are required. The intraflagellar transport(IFT) protein IFT140 is a core component of the IFT system that functions in signaling transduction through the primary cilia. It is reportedly expressed in mesenchymal stem cells and plays a role in bone formation. In this study, we demonstrated that IFT140 was intensively expressed in K14^(+)stem/progenitor cells during the developmental period and early regeneration stage following ligation-induced injuries in murine submandibular glands. In addition, we demonstrated that IFT140^(+)/K14^(+)could self-renew and differentiate into granular duct cells at the developmental stage in vivo. The conditional deletion of Ift140 from K14^(+)cells caused abnormal epithelial structure and function during salivary gland development and inhibited regeneration. IFT140 partly coordinated the function of K14^(+)stem/progenitor cells by modulating ciliary membrane trafficking. Our investigation identified a combined marker, IFT140^(+)/K14^(+), for salivary gland stem/progenitor cells and elucidated the essential role of IFT140 and cilia in regulating salivary stem/progenitor cell differentiation and gland regeneration.

关键词： salivary progenitor injuries

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The role of periodontal disease in atherosclerotic cardiovascular disease

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BIOCELL 2023年第7期47卷 1431-1438页

作者： XIWEI ZHAO JINSONG WANG YIFAN XU JIAN ZHOU LEI HU Beijing Laboratory of Oral Health School of Basic MedicineSchool of StomatologyCapital Medical UniversityBeijing100069China Department of Biochemistry and Molecular Biology School of Basic MedicineCapital Medical UniversityBeijing100069China Immunology research Center for Oral and Systemic Health Beijing Friendship HospitalCapital Medical UniversityBeijing100050China Research Unit of Tooth Development and Regeneration Chinese Academy of Medical SciencesBeijing100700China Department of VIP Dental Service School of StomatologyCapital Medical UniversityBeijing100050China

Atherosclerotic cardiovascular disease(ASCVD)includes a group of disorders of the heart and blood vessels and accounts for major morbidity and premature death worldwide.Periodontitis is a chronic inflammatory disease with the gradual destruction of supporting tissues around the teeth,including gingiva,periodontal ligament,alveolar bone,and cementum.Periodontitis has been found to potentially increase the risk of ASCVD.Generally,oral microorganisms and inflammation are the major factors for periodontitis to the incidence of ASCVD.Recently,evidence has shown that the loss of masticatory function is another important factor of periodontitis to the incidence of ASCVD.In this review,we illustrate the recent finding of the relationship between periodontitis and ASCVD,from a microscale perspective-oral microorganisms,inflammation,and tooth loss.With the high prevalence of periodontitis,it is important to add oral therapy as a regular ASCVD prevention strategy.Regular dental visits could be a helpful strategy for ASCVD patients or general medical practitioners.

关键词： Oral microorganisms Chronic infection tooth loss Periodontitis Atherosclerotic cardiovascular disease

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Dendritic shrinkage after injury: a cellular killer or a necessity for axonal regeneration?

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Neural regeneration research 2019年第8期14卷 1313-1316页

作者： An Beckers Lieve Moons Neural Circuit development and regeneration research Group Department of BiologyKU Leuven

Dendrites form an essential component of the neuronal circuit have been largely overlooked in regenerative research. Nevertheless, subtle changes in the dendritic arbors of neurons are one of the first stages of various neurodegenerative diseases, leading to dysfunctional neuronal networks and ultimately cellular death. Maintaining dendrites is therefore considered an essential neuroprotective strategy. This mini-review aims to discuss an intriguing hypothesis, which postulates that dendritic shrinkage is an important stimulant to boost axonal regeneration, and thus that preserving dendrites might not be the ideal therapeutic method to regain a full functional network upon central nervous system damage. Indeed, our study in zebrafish, a versatile animal model with robust regenerative capacity recently unraveled that dendritic retraction is evoked prior to axonal regrowth after optic nerve injury. Strikingly, inhibiting dendritic pruning upon damage perturbed axonal regeneration. This constraining effect of dendrites on axonal regrowth has sporadically been proposed in literature, as summarized in this short narrative. In addition, the review discusses a plausible underlying mechanism for the observed antagonistic axon-dendrite interplay, which is based on energy restriction inside neurons. Axonal injury indeed leads to a high local energy demand in which efficient axonal energy supply is fundamental to ensure regrowth. At the same time, axonal lesion is known to induce m让ochondrial depolarization, causing energy depletion in the axonal compartment of damaged neurons. Mitochondria, however, become mostly stationary after development, which has been proposed as a potential underlying reason for the low regenerative capacity of adult mammals. Per contra, upon reduced neuronal activity, mitochondrial mobility enhances. In this view, dendritic shrinkage after axonal injury in zebrafish could result in less synaptic input and hence, a release of mitochondria within the soma-dendrite compartment that then translocate to

关键词： axonal regeneration dendritic remodeling retina central nervous system zebrafish mitochondrial transport mitochondrial dynamics energy supply

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人源间充质干细胞药物的研发和转化应用

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Science Bulletin 2023年第9期68卷 860-863页

作者：刘怡 Dana T.Graves 王松灵 Laboratory of Tissue regeneration and Immunology and Department of Periodontics Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionSchool of StomatologyCapital Medical UniversityBeijing 100050China Beijing Laboratory of Oral Health Capital Medical UniversityBeijing 100069China Department of Periodontics School of Dental MedicineUniversity of PennsylvaniaPhiladelphia PA 19104USA Department of Biochemistry and Molecular Biology School of Basic Medical SciencesCapital Medical UniversityBeijing 100069China

In recent years,mesenchymal stem cells(MSCs)have been widely applied in the treatment of various clinical diseases because of multiple advantages and significant therapeutic effects.Despite the long research time and complicated approval procedures,MSC therapy has been gradually progressing and gaining popularity worldwide,including in China.

关键词：间充质干细胞 clinical diseases

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