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Blockade of PD-L1/PD-1 signaling promotes osteo-/odontogenic differentiation through Ras activation

International Journal of Oral Science 2022年第2期14卷 164-173页

作者： So Mi Jeon Je Sun Lim Su Hwan Park hyung Joon kim hyung-ryong kim Jong-Ho Lee Department of Health Sciences The Graduate School of Dong-A UniversityBusanRepublic of Korea Department of Oral Physiology Periodontal Diseases Signaling Network Research CenterDental and Life Science InstituteSchool of DentistryPusan National UniversityYangsanRepublic of Korea Department of Pharmacology College of DentistryJeonbuk National UniversityJeonjuRepublic of Korea Department of Biomedical Sciences Dong-A UniversityBusanRepublic of Korea

The programmed cell death ligand 1(PD-L1)and its receptor programmed cell death 1(PD-1)deliver inhibitory signals to regulate immunological tolerance during immune-mediated diseases.However,the role of PD-1 signaling and its blockade effect on human dental pulp stem cells(h DPSCs)differentiation into the osteo-/odontogenic lineage remain unknown.We show here that PD-L1 expression,but not PD-1,is downregulated during osteo-/odontogenic differentiation of h DPSCs.Importantly,PD-L1/PD-1 signaling has been shown to negatively regulate the osteo-/odontogenic differentiation of h DPSCs.Mechanistically,depletion of either PD-L1 or PD-1 expression increased ERK and AKT phosphorylation levels through the upregulation of Ras enzyme activity,which plays a pivotal role during h DPSCs osteo-/odontogenic differentiation.Treatment with nivolumab(a human anti-PD-1 monoclonal antibody),which targets PD-1 to prevent PD-L1 binding,successfully enhanced osteo-/odontogenic differentiation of h DPSCs through enhanced Ras activity-mediated phosphorylation of ERK and AKT.Our findings underscore that downregulation of PD-L1 expression accompanies during osteo-/odontogenic differentiation,and h DPSCs-intrinsic PD-1 signaling inhibits osteo-/odontogenic differentiation.These findings provide a significant basis that PD-1 blockade could be effective immunotherapeutic strategies in h DPSCs-mediated dental pulp regeneration.

关键词： death finding monoclonal

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Electrically controlled mRNA delivery using a polypyrrolegraphene oxide hybrid film to promote osteogenic differentiation of human mesenchymal stem cells

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Nano Research 2022年第10期15卷 9253-9263页

作者： Huijung kim Kübra Solak Yoojoong Han Yeon-Woo Cho Kyeong-Mo Koo Chang-Dae kim Zhengtang Luo hyungbin Son hyung-ryong kim Ahmet Mavi Tae-hyung kim School of Integrative Engineering Chung-Ang University84 Heukseuk-roDongjak-guSeoul 06974Republic of Korea Department of Nanoscience and Nanoengineering Graduate School of Natural and Applied SciencesAtatürk UniversityErzurum 25240Turkey Department of Chemical and Biological Engineering Hong Kong University of Science and TechnologyKowloonHong Kong 999077China Department of Pharmacology College of DentistryJeonbuk National UniversityJeonju 54896Republic of Korea Department of Nanoscience and Nanoengineering Institute of ScienceAtatürk UniversityErzurum 25240Turkey Department of Mathematics and Science Education Education Faculty of Kazım KarabekirAtatürk UniversityErzurum 25240Turkey

Direct messenger ribonucleic acid(mRNA)delivery to target cells or tissues has revolutionized the field of biotechnology.However,the applicability of regenerative medicine is limited by the technical difficulties of various mRNA-loaded nanocarriers.Herein,we report a new conductive hybrid film that could guide osteogenic differentiation of human adipose-derived mesenchymal stem cells(hADMSCs)via electrically controlled mRNA delivery.To find optimal electrical conductivity and mRNAloading capacity,the polypyrrole-graphene oxide(PPy-GO)hybrid film was electropolymerized on indium tin oxide substrates.We found that the fluorescein sodium salt,a molecule partially mimicking the physical and chemical properties of mRNAs,can be effectively absorbed and released by electrical stimulation(ES).The hADMSCs cultivated on the PPy-GO hybrid film loaded with pre-osteogenic mRNAs showed the highest osteogenic differentiation under electrical stimulation.This platform can load various types of RNAs thus highly promising as a new nucleic acid delivery tool for the development of stem cell-based therapeutics.

关键词： messenger ribonucleic acid(mRNA)delivery polypyrrole graphene oxide electrical release mesenchymal stem cells

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Sphingosine-1-phosphate hinders the osteogenic differentiation of dental pulp stem cells in association with AKT signaling pathways

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International Journal of Oral Science 2022年第2期14卷 192-200页

作者： Bongkun Choi Ji-Eun kim Si-On Park Eun-Young kim Soyoon Oh Hyuksu Choi Dohee Yoon Hyo-Jin Min hyung-ryong kim Eun-Ju Chang Department of Biomedical Sciences Asan Medical CenterUniversity of Ulsan College of MedicineSeoulKorea Stem Cell Immunomodulation Research Center Asan Medical CenterUniversity of Ulsan College of MedicineSeoulKorea Department of Pharmacology College of DentistryJeonbuk National UniversityJeonjuKorea Department of Biochemistry and Molecular Biology Asan Medical CenterUniversity of Ulsan College of MedicineSeoulKorea

Sphingosine-1-phosphate(S1P)is an important lipid mediator that regulates a diverse range of intracellular cell signaling pathways that are relevant to tissue engineering and regenerative medicine.However,the precise function of S1P in dental pulp stem cells(DPSCs)and its osteogenic differentiation remains unclear.We here investigated the function of S1P/S1P receptor(S1PR)-mediated cellular signaling in the osteogenic differentiation of DPSCs and clarified the fundamental signaling pathway.Our results showed that S1P-treated DPSCs exhibited a low rate of differentiation toward the osteogenic phenotype in association with a marked reduction in osteogenesis-related gene expression and AKT activation.Of note,both S1PR1/S1PR3 and S1PR2 agonists significantly downregulated the expression of osteogenic genes and suppressed AKT activation,resulting in an attenuated osteogenic capacity of DPSCs.Most importantly,an AKT activator completely abrogated the S1P-mediated downregulation of osteoblastic markers and partially prevented S1P-mediated attenuation effects during osteogenesis.Intriguingly,the pro-inflammatory TNF-αcytokine promoted the infiltration of macrophages toward DPSCs and induced S1P production in both DPSCs and macrophages.Our findings indicate that the elevation of S1P under inflammatory conditions suppresses the osteogenic capacity of the DPSCs responsible for regenerative endodontics.

关键词： pulp hinder precise

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3D cellular visualization of intact mouse tooth using optical clearing without decalcification

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International Journal of Oral Science 2019年第3期11卷 223-230页

作者： Sujung Hong Jingu Lee Jin Man kim Sun-Young kim hyung-ryong kim Pilhan kim Graduate School of Nanoscience and Technology Korea Advanced Institute of Science and Technology (KAIST)291 Daehak-roYuseong-guRepublic of Korea KI for Health Science and Technology (KIHST) Korea Advanced Institute of Science and Technology (KAIST)291 Deahak-roYuseong-guRepublic of Korea Department of Dentistry CHA Bundang Medical CenterCHA UniversitySeongnamRepublic of Korea Department of Conservative Dentistry and Dental Research Institute Seoul National University School of Dentistry101 Daehak-roJongno-guRepublic of Korea College of Dentistry Institute of Tissue Regeneration Engineering (ITREN)Dankook UniversityCheonanRepublic of Korea Graduate School of Medical Science and Engineering Korea Advanced Institute of Science and Technology (KAIST)291 Daehak-roYuseong-guRepublic of Korea

Dental pulp is composed of nerves,blood vessels,and various types of cells and surrounded by a thick and hard enamel-dentin matrix.Due to its importance in the maintenance of tooth vitality,there have been intensive efforts to analyze the complex cellularlevel organization of the dental pulp in teeth.Although conventional histologic analysis has provided microscopic images of the dental pulp,3-dimensional (3D) cellular-level visualization of the whole dental pulp in an intact tooth has remained a technically challenging task.This is mainly due to the inevitable disruption and loss of microscopic structural features during the process of mechanical sectioning required for the preparation of the tooth sample for histological observation.To accomplish 3D microscopic observation of thick intact tissue,various optical clearing techniques have been developed mostly for soft tissue,and their application for hard tissues such as bone and teeth has only recently started to be investigated.In this work,we established a simple and rapid optical clearing technique for intact mouse teeth without the time-consuming process of decalcification.We achieved 3D cellular-level visualization of the microvasculature and various immune cell distributions in the whole dental pulp of mouse teeth under normal and pathologic conditions.This technique could be used to enable diverse research methods on tooth development and regeneration by providing 3D visualization of various pulpal cells in intact mouse teeth.

关键词： blood vessels Dental pulp soft tissue

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Targeting the RAS/RAF/MAPK pathway for cancer therapy: from mechanism to clinical studies

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Signal Transduction and Targeted Therapy 2024年第1期9卷 173-210页

作者： Md Entaz Bahar Hyun Joon kim Deok ryong kim Department of Biochemistry and Convergence Medical Sciences and Institute of Medical Science Gyeongsang National UniversityCollege of MedicineJinjuSouth Korea Department of Anatomy and Convergence Medical Sciences and Institute of Medical Science Gyeongsang National UniversityCollege of MedicineJinjuSouth Korea

Metastatic dissemination of solid tumors,a leading cause of cancer-related mortality,underscores the urgent need for enhanced insights into the molecular and cellular mechanisms underlying metastasis,chemoresistance,and the mechanistic backgrounds of individuals whose cancers are prone to migration.The most prevalent signaling cascade governed by multi-kinase inhibitors is the mitogen-activated protein kinase(MAPK)pathway,encompassing the RAS–RAF–MAPK kinase(MEK)–extracellular signal-related kinase(ERK)pathway.RAF kinase is a primary mediator of the MAPK pathway,responsible for the sequential activation of downstream targets,such as MEK and the transcription factor ERK,which control numerous cellular and physiological processes,including organism development,cell cycle control,cell proliferation and differentiation,cell survival,and death.Defects in this signaling cascade are associated with diseases such as cancer.RAF inhibitors(RAFi)combined with MEK blockers represent an FDAapproved therapeutic strategy for numerous RAF-mutant cancers,including melanoma,non-small cell lung carcinoma,and thyroid cancer.However,the development of therapy resistance by cancer cells remains an important barrier.Autophagy,an intracellular lysosome-dependent catabolic recycling process,plays a critical role in the development of RAFi resistance in cancer.Thus,targeting RAF and autophagy could be novel treatment strategies for RAF-mutant cancers.In this review,we delve deeper into the mechanistic insights surrounding RAF kinase signaling in tumorigenesis and RAFi-resistance.Furthermore,we explore and discuss the ongoing development of next-generation RAF inhibitors with enhanced therapeutic profiles.Additionally,this review sheds light on the functional interplay between RAF-targeted therapies and autophagy in cancer.

关键词： RAF insight mechanism

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Pedicled abdominal flap using deep inferior epigastric artery perforators for forearm reconstruction: A case report

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World Journal of Clinical Cases 2024年第4期12卷 828-834页

作者： Jae hyung Jeon Kyung Wook kim Hong Bae Jeon Department of Plastic and Reconstructive Surgery Dankook University HospitalCheonan 31116ChungnamSouth Korea Department of Orthopaedic Surgery Dankook University HospitalDankook University College of MedicineCheonan 31116ChungnamSouth Korea Department of Plastic and Reconstructive Surgery Dankook University HospitalDankook University College of MedicineCheonan 31116ChungnamSouth Korea

BACKGROUND Pedicled abdominal flaps are a widely used surgical technique for forearm reconstruction in patients with soft tissue defects.However,some drawbacks include restricted flap size,partial flap loss,and donor-site morbidity.To address these concerns,we present a case of a pedicled abdominal flap using the deep inferior epigastric artery perforators(DIEP)for forearm reconstruction in a patient with a large soft tissue defect.CASE SUMMARY A 46-year-old male patient was admitted to our hospital with forearm injury caused by a pressing machine.A 15 cm×10 cm soft tissue defect with complete rupture of the ulnar side structures of the forearm was found.One week after orthopedic management of the neurovascular injury and fractures using the first stage of Masquelet technique,the patient was referred to the plastic and recon-structive surgery department for wound coverage.Surgical debridement and negative-pressure wound therapy revealed a 20 cm×15 cm soft tissue defect.A pedicle abdominal flap with the DIEP was used to cover the defect.Three weeks later,the flap was detached from the abdomen,and the abdominal defect was directly closed.Subsequently,the second stage of Masquelet technique was performed at the fracture site at week 10.Finally,all donor and recipient sites healed without complications,such as flap dehiscence,infection,hematoma,or necrosis.Fracture site osteosynthesis was achieved without complications.CONCLUSION Pedicled abdominal flap using the DIEP provides a reliable option for forearm reconstruction in patients with large soft tissue defects.

关键词： Forearm injury Open fracture reduction Perforator flap Deep inferior epigastric artery perforators Case report

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Boosting the catalytic activity toward oxygen reduction via a heterostructure of porous iron oxide-decorated 2D NiO/NG nanosheets

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Journal of Energy Chemistry 2024年第6期93卷 669-681,I0016页

作者： Kakali Maiti Matthew T.Curnan hyung Jun kim Kyeounghak kim Jeong Woo Han Department of Chemical Engineering Pohang University of Science and Technology(POSTECH)PohangGyeongbuk 37673Republic of Korea Department of Materials Science and Engineering Research Institute of Advanced MaterialsSeoul National UniversitySeoul 08826Republic of Korea Department of Chemical Engineering Clean-Energy Research InstituteHanyang UniversitySeoul 04763Republic of Korea CSIR-Institute of Minerals and Materials Technology RRL CampusAcharya ViharBhubaneswarOdisha 751013India

As a noble metal substitute,two-dimensional(2D)hierarchical nano-frame structures have attracted great interest as candidate catalysts due to their remarkable advantages-high intrinsic activity,high electron mobility,and straightforward surface functionalization.Therefore,they may replace Pt-based catalysts in oxygen reduction reaction(ORR)applications.Herein,a simple method is developed to design hierarchical nano-frame structures assembled via 2D NiO and N-doped graphene(NG)nanosheets.This procedure can yield nanostructures that satisfy the criteria correlated with improved electrocatalytic performance,such as large surface area,numerous undercoordinated atoms,and high defect densities.Further,porous NG nanosheet architectures,featuring NiO nanosheets densely coordinated with accessible holey Fe_(2)O_(3) moieties,can enhance mesoporosity and balance hydrophilicity.Such improvements can facilitate charge transport and expose formerly inaccessible reaction sites,maximizing active site density utilization.Density functional theory(DFT)calculations reveal favored O_(2) adsorption and dissociation on Fe_(2)O_(3) hybrid structures when supported by 2D NiO and NG nanomaterials,given 2D materials donated charge to Fe_(2)O_(3) active sites.Our systematic studies reveal that synergistic contributions are responsible for enriching the catalytic activity of Fe_(2)O_(3)@NiO/NG in alkaline media-encompassing internal voids and pores,unique hierarchical support structures,and concentrated N-dopant and bimetallic atomic interactions.Ultimately,this work expands the toolbox for designing and synthesizing highly efficient 2D/2D shelled functional nanomaterials with transition metals,endeavoring to benefit energy conversion and related ORR applications.

关键词： N-doped graphene Holey Fe_(2)O_(3)nanocrystals NiO nanosheets High catalytic performance ORR

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Je-Chun-Jun induced apoptosis of human cervical carcinoma HeLa cells

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Acta Pharmacologica Sinica 2004年第10期 132-139页

作者： Han-jung CHAE,Kyung-mi PARK,Geun-youn LEE,Gi-seup JEONG,hyung-rae PARK,hyung-min kim,Soo-wan CHAE,Shim-keun YOO,hyung-ryong kim Iksan Chonbuk 470-739 South Korea Department of Dental Pharmacology and Nano-Science & Technology Research Institute School of Dentistry Wonkwang University Iksan Chonbuk 570-749 South Korea Department of Dental Pharmacology and Nano-Science & Technology Research Institute Department of Pharmacology and the Institute of Cardiovascular Research School of Dentistry Wonkwang University Iksan Chonbuk 570-749 South Korea Department of Dental Pharmacology and Nano-Science & Technology Research Institute School of Dentistry Wonkwang University Iksan Chonbuk 570-749 School of Medicine South Korea Department of Pharmacology College of Oriental Medicine Kyung Hee University Seoul South Korea Department of Pharmacology and the Institute of Cardiovascular Research School of Medicine Chonbuk National University Jeonju Chonbuk National University Chonbuk 464-701 South Korea Department of Gynecology School of Oriental Medicine Wonkwang University Iksan Chonbuk 470-739 South Korea Department of Dental Pharmacology and Nano-Science & Technology Research Institute School of Dentistry Jeonju Wonkwang University Iksan Chonbuk 570-749 South Korea Chonbuk 464-701 South Korea Department of Gynecology School of Oriental Medicine Wonkwang University

AIM: To study the mechanism of Je-Chun-Jun (JCJ)-inducing the apoptosis of the human cervical carcinoma, HeLa cells. METHODS: The cell viability was assessed using MTT assay. The optical density was measured at 570 nm. The caspase activity was measured using 50 mmol/L of fluorogenic substrate, AC-DEVD-AMC (caspase-3), AC-VEID-AMC (caspase-8) or AC-LEHD-AFC (caspase-9). To confirm the expression of proteins, Western blotting was performed. To detect the characteristic of apoptosis chromatin condensation, HeLa cells were stained with Hoechst 33258 in the presence of JCJ. For the cell cycle analysis, HeLa cells were incubated with Propidium iodide (PI) solution. Fluorescence intensity of cell cycle was measured using flow cytometry system. RESULTS: The loss of viability occurred following the exposure of 10 g/L JCJ. Cells treated with 10 g/L JCJ for 3 d exhibited the apoptotic morphology (brightly blue-fluorescent condensed nuclei by Hoechst 33258-staining) and the reduction of cell volume. Cells incubated

关键词： herbal medicine apoptosis p38 MAPK HeLa cells

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Neural Wiskott-Aldrich syndrome protein(N-WASP)promotes distant metastasis in pancreatic ductal adenocarcinoma via activation of LOXL2

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Oncology Research 2024年第4期32卷 615-624页

作者： hyung SUN kim YUN SUN LEE SEUNG MYUNG DONG HYO JUNG kim DA EUN LEE HYEON WOONG KANG MYEONG JIN kim JOON SEONG PARK Department of Surgery Gangnam Severance HospitalYonsei University College of MedicineSeoulKorea Department of Surgery Graduate School of Medical ScienceBrain Korea 21 ProjectYonsei University College of MedicineSeoulKorea Gibiome Co. Ltd.Gyeonggi-doKorea

Pancreatic ductal adenocarcinoma(PDAC)is one of the most aggressive solid malignancies.A specific mechanism of its metastasis has not been established.In this study,we investigated whether Neural Wiskott-Aldrich syndrome protein(N-WASP)plays a role in distant metastasis of PDAC.We found that N-WASP is markedly expressed in clinical patients with PDAC.Clinical analysis showed a notably more distant metastatic pattern in the N-WASP-high group compared to the N-WASP-low group.N-WASP was noted to be a novel mediator of epithelialmesenchymal transition(EMT)via gene expression profile studies.Knockdown of N-WASP in pancreatic cancer cells significantly inhibited cell invasion,migration,and EMT.We also observed positive association of lysyl oxidase-like 2(LOXL2)and focal adhesion kinase(FAK)with the N-WASP-mediated response,wherein EMT and invadopodia function were modulated.Both N-WASP and LOXL2 depletion significantly reduced the incidence of liver and lung metastatic lesions in orthotopic mouse models of pancreatic cancer.These results elucidate a novel role for N-WASP signaling associated with LOXL2 in EMT and invadopodia function,with respect to regulation of intercellular communication in tumor cells for promoting pancreatic cancer metastasis.These findings may aid in the development of therapeutic strategies against pancreatic cancer.

关键词： Pancreatic cancer Neural Wiskott-Aldrich syndrome protein(N-WASP)signaling Metastasis Epithelial-mesenchymal transition(EMT) Lysyl oxidase-like 2(LOXL2)

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New insights into ATR inhibition in muscle invasive bladder cancer:The role of apolipoprotein B mRNA editing catalytic subunit 3B

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Oncology Research 2024年第6期32卷 1021-1030页

作者： HYUNHO kim UIJU CHO SOOK HEE HONG hyung SOON PARK IN-HO kim HO JUNG AN BYOUNG YONG SHIM JIN HYOUNG KANG Division of Medical Oncology Department of Internal MedicineSt.Vincent’s HospitalCollege of MedicineThe Catholic University of KoreaSuwonKorea Department of Pathology St.Vincent’s HospitalCollege of MedicineThe Catholic University of KoreaSuwonKorea Division of Medical Oncology Department of Internal MedicineSeoul St.Mary’s HospitalCollege of MedicineThe Catholic University of KoreaSeoulKorea

Background:Apolipoprotein B mRNA editing catalytic polypeptide(APOBEC),an endogenous mutator,induces DNA damage and activates the ataxia telangiectasia and Rad3-related(ATR)-checkpoint kinase 1(Chk1)pathway.Although cisplatin-based therapy is the mainstay for muscle-invasive bladder cancer(MIBC),it has a poor survival rate.Therefore,this study aimed to evaluate the efficacy of an ATR inhibitor combined with cisplatin in the treatment of APOBEC catalytic subunit 3B(APOBEC3B)expressing MIBC.Methods:Immunohistochemical staining was performed to analyze an association between APOBEC3B and ATR in patients with MIBC.The APOBEC3B expression in MIBC cell lines was assessed using real-time polymerase chain reaction and western blot analysis.Western blot analysis was performed to confirm differences in phosphorylated Chk1(pChk1)expression according to the APOBEC3B expression.Cell viability and apoptosis analyses were performed to examine the anti-tumor activity of ATR inhibitors combined with cisplatin.Results:There was a significant association between APOBEC3B and ATR expression in the tumor tissues obtained from patients with MIBC.Cells with higher APOBEC3B expression showed higher pChk1 expression than cells expressing low APOBEC3B levels.Combination treatment of ATR inhibitor and cisplatin inhibited cell growth in MIBC cells with a higher APOBEC3B expression.Compared to cisplatin single treatment,combination treatment induced more apoptotic cell death in the cells with higher APOBEC3B expression.Conclusion:Our study shows that APOBEC3B’s higher expression status can enhance the sensitivity of MIBC to cisplatin upon ATR inhibition.This result provides new insight into appropriate patient selection for the effective application of ATR inhibitors in MIBC.

关键词： Apolipoprotein B mRNA editing catalytic polypeptide(APOBEC) Ataxia telangiectasia and Rad3-related(ATR) Bladder cancer DNA damage response DNA replication stress

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