Upregulation of miR-23b Enhances the Autologous Therapeutic Potential for Degenerative Arthritis by Targeting PRKACB in Synovial Fluid-Derived Mesenchymal Stem Cells from Patients

함온주,이창연,송병욱,이세연,김란,박준희,이지현,서향희,이채연,정용안,맹이소,이민영,김종민,황지환,우동균,장우철 한국분자세포생물학회 2014 Molecules and cells Vol.37 No.6

The use of synovial fluid-derived mesenchymal stem cells (SFMSCs) obtained from patients with degenerative arthropathy may serve as an alternative therapeutic strategy in osteoarthritis (OA) and rheumatoid arthritis (RA). For treatment of OA and RA patients, autologous transplantation of differentiated MSCs has several beneficial effects for cartilage regeneration including immunomodulatory activity. In this study, we induced chondrogenic differentiation of SFMSCs by inhibiting protein kinase A (PKA) with a small molecule and microRNA (miRNA). Chondrogenic differentiation was confirmed by PCR and immunocytochemistry using probes specific for aggrecan, the major cartilaginous proteoglycan gene. Absorbance of alcian blue stain to detect chondrogenic differentiation was increased in H-89 and/or miRNA-23b-transfected cells. Furthermore, expression of matrix meta¬lloproteinase (MMP)-9 and MMP-2 was decreased in treated cells. Therefore, differentiation of SFMSCs into chondrocytes through inhibition of PKA signaling may be a therapeutic option for OA or RA patients.

In vitro Organoid model of human lungs from pluripotent stem cells

강지수,함온주,이영선,이미옥 환경독성보건학회 2022 한국독성학회 심포지움 및 학술발표회 Vol.2022 No.10

TCF4-Targeting miR-124 is Differentially Expressed amongst Dendritic Cell Subsets

Sun Murray Han,나혜영,함온주,최완호,손모아,류슬혜,인현주,황기철,박채규 대한면역학회 2016 Immune Network Vol.16 No.1

Dendritic cells (DCs) are professional antigen-presenting cells that sample their environment and present antigens to naïve T lymphocytes for the subsequent antigen-specific immune responses. DCs exist in a range of distinct subpopulations including plasmacytoid DCs (pDCs) and classical DCs (cDCs), with the latter consisting of the cDC1 and cDC2 lineages. Although the roles of DC-specific transcription factors across the DC subsets have become understood, the posttranscriptional mechanisms that regulate DC development are yet to be elucidated. MicroRNAs (miRNAs) are pivotal posttranscriptional regulators of gene expression in a myriad of biological processes, but their contribution to the immune system is just beginning to surface. In this study, our in-house probe collection was screened to identify miRNAs possibly involved in DC development and function by targeting the transcripts of relevant mouse transcription factors. Examination of DC subsets from the culture of mouse bone marrow with Flt3 ligand identified high expression of miR-124 which was able to target the transcript of TCF4, a transcription factor critical for the development and homeostasis of pDCs. Further expression profiling of mouse DC subsets isolated from in vitro culture as well as via ex vivo purification demonstrated that miR-124 was outstandingly expressed in CD24 + cDC1 cells compared to in pDCs and CD172α+ cDC2 cells. These results imply that miR-124 is likely involved in the processes of DC subset development by posttranscriptional regulation of a transcription factor(s).

Novel Lactam Type Pyridine Derivatives Improves Myocardium Dysfunction Derived from Ischemic Injury

김경은,최은주,장양수,송희상,차민지,송병욱,함온주,이장연,최성용,이세연,황기철 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13

The extended acute myocardial ischemia (AMI) results in cardiac myocytes death. In the present study, we show that lactam pyridine derivative, SK-D80375, have the effects on cell survival in hypoxic cardiomyocytes and might be used as an anti-ischemic drug. The lactam pyridine derivatives are inhibitors of the late sodium current, which decreases sodium-dependent intracellular calcium overload in ischemia/reperfusion-injured hearts. We found that pretreatment with SK-D80375 significantly decreased the level of intracellular Ca2+ and the expression level of the Na+-Ca2+ exchanger by 39±2.5% and 19±0.5%, respectively in hypoxic cardiomyocytes compared to untreated controls. In addition, the expression level of sarcoplasmic reticulum Ca2+ ATPase 2a was significantly increased by 37±1.5% in SK-D80375-treated hypoxic cardiomyocytes compared to untreated controls. The induction of Hsp70 was observed in SK-D80375-treated hypoxic cardiomyocytes with dose-dependent manner and the highest level of Hsp70 was induced at the concentration of 2.5 μM SK-D80375. The echocardiographic analysis showed that heart function was significantly improved in SK-D80375-injected ischemic hearts. These results demonstrate that lactam pyridine derivative, SK-D80375, have beneficial effects on hypoxia-induced cell death, therefore, might be used as a novel anti-ischemia drug.

Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 TargetedmicroRNA- 221

장우철,이소명,김란,박상인,정유진,함온주,이지현,김지형,오세경,이민영,김종민,박문서,정용안,황기철,맹이소 한국분자세포생물학회 2015 Molecules and cells Vol.38 No.7

The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule- 1 was upregulated by microRNA-221 in mesenchymalstem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targetedintercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

저산소 상태의 백서 심근세포에서 Regucalcin의 특이적 발현 및 역할

임소연,최은주,장양수,송병욱,차민지,이창연,함온주,최성용,황기철,이세연 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13

Regucalcin (SMP30) has been proposed to be involved in the maintenance of calcium homeostasis. Although the expression of regucalcin was regulated in the liver and kidney during the embryogenesis and maturation of these tissues, the roles of regucalcin were not defined yet in heart. This study focused on the investigation of the differential expression changes in regucalcin and its function in hypoxic cardiomyocytes. The expression level of regucalcin was the highest in 7 days after neonatal stage of rat heart. In hypoxic condition, reactive oxygen species (ROS) production and calcium level were decreased in regucalcin-overexpressed cardiomyocytes about 60% compared to normal cells. Regucalcin-transfected cells were slowly induced cell death in H2O2 treated condition and not reduced proliferation in hypoxic condition. In proliferation-related signals, PKC-MEK1/2-ERK1/2 activation of regucalcin-overexpressed cells was recovered in hypoxia, compared to hypoxic cardiomyocytes, and expression of proto-oncogene was only affected c-myc by regucalcin. Transfected cardiomyocytes demonstrated that apoptotic signal was hampered in regucalcin-regulated manner. In conclusion, these data suggest that regucalcin may regulate proliferation and cell death in cardiomyocytes via calcium homeostasis. Regucalcin (SMP30) has been proposed to be involved in the maintenance of calcium homeostasis. Although the expression of regucalcin was regulated in the liver and kidney during the embryogenesis and maturation of these tissues, the roles of regucalcin were not defined yet in heart. This study focused on the investigation of the differential expression changes in regucalcin and its function in hypoxic cardiomyocytes. The expression level of regucalcin was the highest in 7 days after neonatal stage of rat heart. In hypoxic condition, reactive oxygen species (ROS) production and calcium level were decreased in regucalcin-overexpressed cardiomyocytes about 60% compared to normal cells. Regucalcin-transfected cells were slowly induced cell death in H2O2 treated condition and not reduced proliferation in hypoxic condition. In proliferation-related signals, PKC-MEK1/2-ERK1/2 activation of regucalcin-overexpressed cells was recovered in hypoxia, compared to hypoxic cardiomyocytes, and expression of proto-oncogene was only affected c-myc by regucalcin. Transfected cardiomyocytes demonstrated that apoptotic signal was hampered in regucalcin-regulated manner. In conclusion, these data suggest that regucalcin may regulate proliferation and cell death in cardiomyocytes via calcium homeostasis.