Publications by authors named "Jung-Woong Kim"

64 Publications

A Fully Human Monoclonal Antibody Targeting cKIT Is a Potent Inhibitor of Pathological Choroidal Neovascularization in Mice.

Pharmaceutics 2021 Aug 20;13(8). Epub 2021 Aug 20.

Department of Global Innovative Drug, Graduate School of Chung-Ang University, Seoul 06974, Korea.

Stem cell factor (SCF) and its receptor, cKIT, are novel regulators of pathological neovascularization in the eye, which suggests that inhibition of SCF/cKIT signaling may be a novel pharmacological strategy for treating neovascular age-related macular degeneration (AMD). This study evaluated the therapeutic potential of a newly developed fully human monoclonal antibody targeting cKIT, NN2101, in a murine model of neovascular AMD. In hypoxic human endothelial cells, NN2101 substantially inhibited the SCF-induced increase in angiogenesis and activation of the cKIT signaling pathway. In a murine model of neovascular AMD, intravitreal injection of NN2101 substantially inhibited the SCF/cKIT-mediated choroidal neovascularization (CNV), with efficacy comparable to aflibercept, a vascular endothelial growth factor inhibitor. A combined intravitreal injection of NN2101 and aflibercept resulted in an additive therapeutic effect on CNV. NN2101 neither caused ocular toxicity nor interfered with the early retinal vascular development in mice. Ocular pharmacokinetic analysis in rabbits indicated that NN2101 demonstrated a pharmacokinetic profile suitable for intravitreal injection. These findings provide the first evidence of the potential use of the anti-cKIT blocking antibody, NN2101, as an alternative or additive therapeutic for the treatment of neovascular AMD.
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http://dx.doi.org/10.3390/pharmaceutics13081308DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8400730PMC
August 2021

Proneural genes define ground-state rules to regulate neurogenic patterning and cortical folding.

Neuron 2021 Sep 17;109(18):2847-2863.e11. Epub 2021 Aug 17.

Department of Medical Genetics, ACHRI, HBI, University of Calgary, Calgary, AB T2N 4N1, Canada.

Asymmetric neuronal expansion is thought to drive evolutionary transitions between lissencephalic and gyrencephalic cerebral cortices. We report that Neurog2 and Ascl1 proneural genes together sustain neurogenic continuity and lissencephaly in rodent cortices. Using transgenic reporter mice and human cerebral organoids, we found that Neurog2 and Ascl1 expression defines a continuum of four lineage-biased neural progenitor cell (NPC) pools. Double NPCs, at the hierarchical apex, are least lineage restricted due to Neurog2-Ascl1 cross-repression and display unique features of multipotency (more open chromatin, complex gene regulatory network, G pausing). Strikingly, selectively eliminating double NPCs by crossing Neurog2-Ascl1 split-Cre mice with diphtheria toxin-dependent "deleter" strains locally disrupts Notch signaling, perturbs neurogenic symmetry, and triggers cortical folding. In support of our discovery that double NPCs are Notch-ligand-expressing "niche" cells that control neurogenic periodicity and cortical folding, NEUROG2, ASCL1, and HES1 transcript distribution is modular (adjacent high/low zones) in gyrencephalic macaque cortices, prefiguring future folds.
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http://dx.doi.org/10.1016/j.neuron.2021.07.007DOI Listing
September 2021

Anti-inflammatory effect of Ailanthus altissima (Mill.) Swingle leaves in lipopolysaccharide-stimulated astrocytes.

J Ethnopharmacol 2021 Jul 13:114258. Epub 2021 Jul 13.

Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul, 06974, Republic of Korea. Electronic address:

Ethnopharmacological Relevance: Activated astrocytes are involved in the progression of neurodegenerative diseases. Traditionally, Ailanthus altissima (Mill.) Swingle, widely distributed in East Asia, has been used as a medicine for the treatment of fever, gastric diseases, and inflammation. Although A. altissima has been reported to play an anti-inflammatory role in peripheral tissues or cells, its role in the central nervous system (CNS) remains unclear.

Aim Of The Study: In the present study, we investigated the anti-inflammatory effects and mechanism of action of A. altissima in primary astrocytes stimulated by lipopolysaccharide (LPS).

Materials And Methods: A nitrite assay was used to measure nitric oxide (NO) production, and the tetrazolium salt 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to determine cytotoxicity. The expression levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and mitogen-activated protein kinase (MAPK) were determined with western blotting. Reverse-transcription PCR was used to assess the expression of inflammatory cytokines. The levels of reactive oxygen species were measured using 2,7-dichlorodihydrofluorescein diacetate. Luciferase assay and immunocytochemistry were used for assessing nuclear factor-kappa B (NF-κB) transcription and p65 localization, respectively. Memory and social interaction were analyzed using the Y-maze and three-chamber tests, respectively.

Results: The ethanol extract of A. altissima leaves (AAE) inhibited iNOS and COX-2 expression in LPS-stimulated astrocytes. Moreover, AAE reduced the transcription of various proinflammatory mediators, hindered NF-κB activation, and suppressed extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) activation without p38 activation. Ultra-high performance liquid chromatography with mass spectrometry analysis revealed that AAE comprised ethyl gallate, quercetin, and kaempferol, along with luteolin, which has anti-inflammatory properties, and repressed LPS-induced nitrite levels and the nuclear translocation of p65. Finally, oral administration of AAE attenuated LPS-induced memory and social impairment in mice and repressed LPS-induced ERK and JNK activation in the cortices of mice.

Conclusion: AAE could have therapeutic uses in the treatment of neuroinflammatory diseases via suppression of astrocyte activation.
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http://dx.doi.org/10.1016/j.jep.2021.114258DOI Listing
July 2021

Microtubule Acetylation Controls MDA-MB-231 Breast Cancer Cell Invasion through the Modulation of Endoplasmic Reticulum Stress.

Int J Mol Sci 2021 Jun 2;22(11). Epub 2021 Jun 2.

Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

During aggressive cancer progression, cancer cells adapt to unique microenvironments by withstanding various cellular stresses, including endoplasmic reticulum (ER) stress. However, the mechanism whereby cancer cells overcome the ER stress to survive remains to be elucidated. Herein, we demonstrated that microtubule acetylation in cancer cells grown on a stiff matrix promotes cancer progression by preventing excessive ER stress. Downregulation of microtubule acetylation using shRNA or CRSIPR/Cas9 techniques targeting , which encodes α-tubulin N-acetyltransferase (αTAT1), resulted in the upregulation of ER stress markers, changes in ER morphology, and enhanced tunicamycin-induced UPR signaling in cancer cells. A set of genes involved in cancer progression, especially focal adhesion genes, were downregulated in both -knockout and tunicamycin-treated cells, whereas overexpression restored the gene expression inhibited by tunicamycin. Finally, the expression of and ER stress marker genes were negatively correlated in various breast cancer types. Taken together, our results suggest that disruption of microtubule acetylation is a potent therapeutic tool for preventing breast cancer progression through the upregulation of ER stress. Moreover, and ER stress marker genes may be useful diagnostic markers in various breast cancer types.
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http://dx.doi.org/10.3390/ijms22116018DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8199658PMC
June 2021

Transcriptomic profiling of three-dimensional cholangiocyte spheroids long term exposed to repetitive Clonorchis sinensis excretory-secretory products.

Parasit Vectors 2021 Apr 20;14(1):213. Epub 2021 Apr 20.

Department of Convergence Medicine, University of Ulsan College of Medicine and Asan Institute for Life Sciences, Asan Medical Center, Seoul, 05505, Republic of Korea.

Background: Biliary tract infection with the carcinogenic human liver fluke, Clonorchis sinensis, provokes chronic inflammation, epithelial hyperplasia, periductal fibrosis, and even cholangiocarcinoma. Complications are proportional to the intensity and duration of the infection. In addition to mechanical irritation of the biliary epithelia from worms, their excretory-secretory products (ESPs) cause chemical irritation, which leads to inflammation, proliferation, and free radical generation.

Methods: A three-dimensional in vitro cholangiocyte spheroid culture model was established, followed by ESP treatment. This allowed us to examine the intrinsic pathological mechanisms of clonorchiasis via the imitation of prolonged and repetitive in vivo infection.

Results: Microarray and RNA-Seq analysis revealed that ESP-treated cholangiocyte H69 spheroids displayed global changes in gene expression compared to untreated spheroids. In ESP-treated H69 spheroids, 185 and 63 probes were found to be significantly upregulated and downregulated, respectively, corresponding to 209 genes (p < 0.01, fold change > 2). RNA-Seq was performed for the validation of the microarray results, and the gene expression patterns in both transcriptome platforms were well matched for 209 significant genes. Gene ontology analysis demonstrated that differentially expressed genes were mainly classified into immune system processes, the extracellular region, and the extracellular matrix. Among the upregulated genes, four genes (XAF1, TRIM22, CXCL10, and BST2) were selected for confirmation using quantitative RT-PCR, resulting in 100% similar expression patterns in microarray and RNA-Seq.

Conclusions: These findings broaden our understanding of the pathological pathways of liver fluke-associated hepatobiliary disorders and suggest a novel therapeutic strategy for this infectious cancer.
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http://dx.doi.org/10.1186/s13071-021-04717-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8056535PMC
April 2021

Mercury Chloride but Not Lead Acetate Causes Apoptotic Cell Death in Human Lung Fibroblast MRC5 Cells via Regulation of Cell Cycle Progression.

Int J Mol Sci 2021 Mar 2;22(5). Epub 2021 Mar 2.

Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

Heavy metals are important for various biological systems, but, in excess, they pose a serious risk to human health. Heavy metals are commonly used in consumer and industrial products. Despite the increasing evidence on the adverse effects of heavy metals, the detailed mechanisms underlying their action on lung cancer progression are still poorly understood. In the present study, we investigated whether heavy metals (mercury chloride and lead acetate) affect cell viability, cell cycle, and apoptotic cell death in human lung fibroblast MRC5 cells. The results showed that mercury chloride arrested the sub-G and G/M phases by inducing cyclin B1 expression. In addition, the exposure to mercury chloride increased apoptosis through the activation of caspase-3. However, lead had no cytotoxic effects on human lung fibroblast MRC5 cells at low concentration. These findings demonstrated that mercury chloride affects the cytotoxicity of MRC5 cells by increasing cell cycle progression and apoptotic cell death.
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http://dx.doi.org/10.3390/ijms22052494DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7958599PMC
March 2021

Transcriptome Analysis Reveals HgCl Induces Apoptotic Cell Death in Human Lung Carcinoma H1299 Cells through Caspase-3-Independent Pathway.

Int J Mol Sci 2021 Feb 18;22(4). Epub 2021 Feb 18.

Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

Mercury is one of the detrimental toxicants that can be found in the environment and exists naturally in different forms; inorganic and organic. Human exposure to inorganic mercury, such as mercury chloride, occurs through air pollution, absorption of food or water, and personal care products. This study aimed to investigate the effect of HgCl on cell viability, cell cycle, apoptotic pathway, and alters of the transcriptome profiles in human non-small cell lung cancer cells, H1299. Our data show that HgCl treatment causes inhibition of cell growth via cell cycle arrest at G/G- and S-phase. In addition, HgCl induces apoptotic cell death through the caspase-3-independent pathway. Comprehensive transcriptome analysis using RNA-seq indicated that cellular nitrogen compound metabolic process, cellular metabolism, and translation for biological processes-related gene sets were significantly up- and downregulated by HgCl treatment. Interestingly, comparative gene expression patterns by RNA-seq indicated that mitochondrial ribosomal proteins were markedly altered by low-dose of HgCl treatment. Altogether, these data show that HgCl induces apoptotic cell death through the dysfunction of mitochondria.
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http://dx.doi.org/10.3390/ijms22042006DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922270PMC
February 2021

Bisphenol A Exposure Changes the Transcriptomic and Proteomic Dynamics of Human Retinoblastoma Y79 Cells.

Genes (Basel) 2021 02 11;12(2). Epub 2021 Feb 11.

Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

Bisphenol A (BPA) is a xenoestrogen chemical commonly used to manufacture polycarbonate plastics and epoxy resin and might affect various human organs. However, the cellular effects of BPA on the eyes have not been widely investigated. This study aimed to investigate the cellular cytotoxicity by BPA exposure on human retinoblastoma cells. BPA did not show cytotoxic effects, such as apoptosis, alterations to cell viability and cell cycle regulation. Comparative analysis of the transcriptome and proteome profiles were investigated after long-term exposure of Y79 cells to low doses of BPA. Transcriptome analysis using RNA-seq revealed that mRNA expression of the post-transcriptional regulation-associated gene sets was significantly upregulated in the BPA-treated group. Cell cycle regulation-associated gene sets were significantly downregulated by exposure to BPA. Interestingly, RNA-seq analysis at the transcript level indicated that alternative splicing events, particularly retained introns, were noticeably altered by low-dose BPA treatment. Additionally, proteome profiling using MALDI-TOF-MS identified a total of nine differentially expressed proteins. These results suggest that alternative splicing events and altered gene/protein expression patterns are critical phenomena affected by long-term low-dose BPA exposure. This represents a novel marker for the detection of various diseases associated with environmental pollutants such as BPA.
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http://dx.doi.org/10.3390/genes12020264DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7918513PMC
February 2021

Ablation of Reduces Cell Viability and Increases UVC-Mediated Apoptosis in Hepatocarcinoma HepG2 Cells.

Genes (Basel) 2021 01 30;12(2). Epub 2021 Jan 30.

Department of Life Science, Chung-Ang University, Seoul 06974, Korea.

Uracil is an unavoidable aberrant base in DNA sequences, the repair of which takes place by a highly efficient base excision repair mechanism. The removal of uracil from the genome requires multiple biochemical steps with conformational changes of DNA that inhibit DNA replication and interfere with transcription. However, the relevance of uracil in DNA for cellular physiology and transcriptional regulation is not fully understood. We investigated the functional roles of using knock-down (KD) and knock-out (KO) models. The proliferation ratio of KD and KO cells was decreased compared to WT control cells, and the cell cycle was arrested in the G2/M phases before the transition to mitosis. The apoptotic cell death was increased in KD and KO cell lines through the increase of BAX and active caspase 3 expression. Phospho-gamma-H2AX expression, which reflected accumulated DNA damage, was also increased in KO cells. Moreover, the apoptotic cells by DNA damage accumulation were markedly increased in KD and KO cells after ultraviolet C irradiation. Transcriptomic analysis using RNA-seq revealed that was involved in gene sets expression including cell cycle transition and chromatin silencing. Together, the results implicate SMUG1 as a critical factor in cell cycle and transcriptional regulation.
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http://dx.doi.org/10.3390/genes12020201DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7911780PMC
January 2021

Acetylation of UHRF1 Regulates Hemi-methylated DNA Binding and Maintenance of Genome-wide DNA Methylation.

Cell Rep 2020 07;32(4):107958

Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul 06974, Republic of Korea. Electronic address:

UHRF1 is a key regulator in DNA methylation maintenance. It binds histone H3K9me2/3 and hemi-methylated DNA and recruits DNMT1 to DNA replication forks during S phase. However, the regulatory mechanism of hemi-methylated DNA binding activity of UHRF1 remains unknown. In this study, we reveal that acetylation of UHRF1 is regulated by PCAF and HDAC1. We show that UHRF1 acetylation at K490 attenuates its binding affinity to hemi-methylated DNA. We analyze genome-wide DNA methylation and gene-expression patterns using stable cell lines and discover that cells where the endogenous UHRF1 is replaced with an acetyl-mimetic (UHRF1 K490Q) mutant show deficiencies in inherited DNA methylation and show different gene-expression patterns in genes related to cell survival. These results reveal that precise regulation of UHRF1 acetylation is required to maintain DNA methylation during cell division and control cell survival.
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http://dx.doi.org/10.1016/j.celrep.2020.107958DOI Listing
July 2020

Simple centrifugal fractionation to reduce the size distribution of cellulose nanofibers.

Sci Rep 2020 07 16;10(1):11744. Epub 2020 Jul 16.

Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, 100 Inha-Ro, Michuhol-Ku, Incheon, 22212, Korea.

Since cellulose nanofiber (CNF) has unique characteristics in terms of renewability, high specific elastic modulus and strength and transparency, it is attractive for a building block of future materials. CNF can be extracted from various natural resource by several means. However, the size of the extracted CNF is very broad and uniformity of the extracted CNF is very important for many applications. Thus, a fractionation process is necessary to obtain a uniformly sized CNF. In this paper, a simple centrifugal fractionation was carried out to reduce the size distribution of the extracted CNF suspension from hardwood pulp by the combination of TEMPO oxidation and aqueous counter collision methods. The original CNF suspension was diluted and centrifuged under low speed to remove cellulose microfibers then centrifuged under high speed to separate very small CNF. The centrifugation condition is 10 k rpm for 1 h followed by 45 k rpm for 4 h. The fractionated CNF was analyzed by an atomic force microscopy, and the length and width distribution histogram analysis was utilized. UV-visible analysis, FT-IR and XRD crystallinity analysis were carried out to analyze all fractionated CNFs and the original CNF. After centrifugal fractionation, the width and length distribution range were reduced by 62% and 70%, respectively. It is shown that the centrifugal fractionation is an easy and efficient method to fractionate a uniform CNF suspension.
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http://dx.doi.org/10.1038/s41598-020-68642-7DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7366905PMC
July 2020

Ethylparaben induces apoptotic cell death in human placenta BeWo cells via the Caspase-3 pathway.

Anim Cells Syst (Seoul) 2020 9;24(1):34-43. Epub 2020 Jan 9.

Department of Life Science, Chung-Ang University, Seoul, South Korea.

Parabens are generally used as preservatives in foods, pharmaceuticals, and various other commercial products. Among them, ethylparaben has weaker estrogenic characteristics than endogenous estrogen. However, growing evidence indicates that ethylparaben has an adverse effect on various human tissues. Here, we investigated whether ethylparaben induces cell death by affecting cell viability, cell proliferation, cell cycle, and apoptosis using the human placenta cell line BeWo. Ethylparaben significantly decreased cell viability in a dose-dependent manner. It caused cell cycle arrest at sub-G1 by reducing the expression of cyclin D1, whereas it decreased the cell proportion at the G0/G1 and S phases. Furthermore, we verified that ethylparaben induces apoptotic cell death by enhancing the activity of Caspase-3. Taken together, our results suggest that ethylparaben exerts cytotoxic effects in human placental BeWo cells via cell cycle arrest and apoptotic pathways.
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http://dx.doi.org/10.1080/19768354.2020.1711804DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7048193PMC
January 2020

Steered Pull Simulation to Determine Nanomechanical Properties of Cellulose Nanofiber.

Materials (Basel) 2020 Feb 5;13(3). Epub 2020 Feb 5.

Creative Research Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, 100 Inha-ro, Michuhol-ku, Incheon 22212, Korea.

Cellulose nanofiber (CNF) exhibits excellent mechanical properties, which has been extensively proven through experimental techniques. However, understanding the mechanisms and the inherent structural behavior of cellulose is important in its vastly growing research areas of applications. This study focuses on taking a look into what happens to the atomic molecular interactions of CNF, mainly hydrogen bond, in the presence of external force. This paper investigates the hydrogen bond disparity within CNF structure. To achieve this, molecular dynamics simulations of cellulose I β nanofibers are carried out in equilibrated conditions in water using GROMACS software in conjunction with OPLS-AA force field. It is noted that the hydrogen bonds within the CNF are disrupted when a pulling force is applied. The simulated Young's modulus of CNF is found to be 161 GPa. A simulated shear within the cellulose chains presents a trend with more hydrogen bond disruptions at higher forces.
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http://dx.doi.org/10.3390/ma13030710DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7041381PMC
February 2020

Effect of Process Orientation on the Mechanical Behavior and Piezoelectricity of Electroactive Paper.

Materials (Basel) 2020 Jan 3;13(1). Epub 2020 Jan 3.

CRC for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, Incheon 22212, Korea.

This paper reports the effect of process orientation on the mechanical behavior and piezoelectricity of electroactive paper (EAPap) made from natural cotton pulp. EAPap is fabricated by a casting and wet drawing of cellulose film after dissolving cotton with LiCl and DMAc solvent. During the fabrication, permanent wrinkles, a possible factor for performance deterioration, were found in the films. Finite element method was introduced to identify the formation mechanism behind the wrinkles. The simulation results show that the wrinkles were caused by buckling and are inevitable under any conditions. The tensile and piezoelectric tests show that the orientation dependency of the stretched EAPap gives the anisotropic characteristics on both mechanical and piezoelectric properties. In this research, the anisotropic elastic moduli and Poisson's ratios are reported. The piezoelectric charge constant of EAPap in the linear elastic is calculated. The piezoelectric charge constants of EAPap are associated with the alignment angle in the order of 45° > 0° > 90° due to the strong shear effect. The higher stretching ratio gives the higher piezoelectricity due to the alignment of the molecular chains and the microstructure in EAPap. The highest piezoelectric charge constant is found to be 12 pC/N at a stretching ratio of 1.6 and aligning angle of 45°.
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http://dx.doi.org/10.3390/ma13010204DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6981422PMC
January 2020

Dynein-mediated nuclear translocation of yes-associated protein through microtubule acetylation controls fibroblast activation.

Cell Mol Life Sci 2020 Oct 7;77(20):4143-4161. Epub 2020 Jan 7.

Department of Life Science, Chung-Ang University, Seoul, 06974, Republic of Korea.

Myofibroblasts are the major cell type that is responsible for increase in the mechanical stiffness in fibrotic tissues. It has well documented that the TGF-β/Smad axis is required for myofibroblast differentiation under the rigid substrate condition. However, the mechanism driving myofibroblast differentiation in soft substrates remains unknown. In this research, we demonstrated that interaction of yes-associated protein (YAP) and acetylated microtubule via dynein, a microtubule motor protein drives nuclear localization of YAP in the soft matrix, which in turn increased TGF-β1-induced transcriptional activity of Smad for myofibroblast differentiation. Pharmacological and genetical disruption of dynein impaired the nuclear translocation of YAP and decreased the TGF-β1-induced Smad activity even though phosphorylation and nuclear localization of Smad occurred normally in α-tubulin acetyltransferase 1 (α-TAT1) knockout cell. Moreover, microtubule acetylation prominently appeared in the fibroblast-like cells nearby the blood vessel in the fibrotic liver induced by CCl administration, which was conversely decreased by TGF-β receptor inhibitor. As a result, quantitative inhibition of microtubule acetylation may be suggested as a new target for overcoming fibrotic diseases.
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http://dx.doi.org/10.1007/s00018-019-03412-xDOI Listing
October 2020

Transcriptome dynamics of alternative splicing events revealed early phase of apoptosis induced by methylparaben in H1299 human lung carcinoma cells.

Arch Toxicol 2020 01 20;94(1):127-140. Epub 2019 Nov 20.

Department of Life Science, Chung-Ang University, Seoul, 06974, South Korea.

Methylparaben is most frequently used as an antimicrobial preservative in pharmaceuticals and foods. Methylparaben has been subjected to toxicological studies owing to the increasing concern regarding its possible impact on the environment and human health. However, the cytotoxicity and underlying mechanisms of methylparaben exposure in human lung cells have not been explored. Here, we investigated the effect of methylparaben on cell cycle, apoptotic pathways, and changes in the transcriptome profiles in human lung cells. Our results demonstrate that treatment with methylparaben causes inhibition of cell growth. In addition, methylparaben induced S- and G2/M-phase arrest as a result of enhanced apoptosis. Transcriptome analysis using RNA-seq revealed that mRNA expression of ER stress- and protein misfolding-related gene sets was upregulated in methylparaben-treated group. RNA splicing- and maturation-related gene sets were significantly down-regulated by methylparaben treatment. Interestingly, RNA-seq analysis at the transcript level revealed that alternative splicing events, especially retained intron, were markedly changed by a low dose of methylparaben treatment. Altogether, these data show that methylparaben induces an early phase of apoptosis through cell cycle arrest and downregulation of mRNA maturation.
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http://dx.doi.org/10.1007/s00204-019-02629-wDOI Listing
January 2020

Induction of Redox-Active Gene Expression by CoCl Ameliorates Oxidative Stress-Mediated Injury of Murine Auditory Cells.

Antioxidants (Basel) 2019 Sep 16;8(9). Epub 2019 Sep 16.

Department of Otolaryngology, University of Ulsan College of Medicine, Asan Medical Center, 88 Olympic-ro 43-gil, Songpa-Gu, Seoul 05505, Korea.

Free radicals formed in the inner ear in response to high-intensity noise, are regarded as detrimental factors for noise-induced hearing loss (NIHL). We reported previously that intraperitoneal injection of cobalt chloride attenuated the loss of sensory hair cells and NIHL in mice. The present study was designed to understand the preconditioning effect of CoCl on oxidative stress-mediated cytotoxicity. Treatment of auditory cells with CoCl promoted cell proliferation, with increases in the expressions of two redox-active transcription factors (hypoxia-inducible factor 1α, HIF-1α, nuclear factor erythroid 2-related factor 2; Nrf-2) and an antioxidant enzyme (peroxiredoxin 6, Prdx6). Hydrogen peroxide treatment resulted in the induction of cell death and reduction of these protein expressions, reversed by pretreatment with CoCl. Knockdown of HIF-1α or Nrf-2 attenuated the preconditioning effect of CoCl. Luciferase reporter analysis with a Prdx6 promoter revealed transactivation of Prdx6 expression by HIF-1α and Nrf-2. The intense immunoreactivities of HIF-1α, Nrf-2, and Prdx6 in the organ of Corti (OC), spiral ganglion cells (SGC), and stria vascularis (SV) of the cochlea in CoCl-injected mice suggested CoCl-induced activation of HIF-1α, Nrf-2, and Prdx6 in vivo. Therefore, we revealed that the protective effect of CoCl is achieved through distinctive signaling mechanisms involving HIF-1α, Nrf-2, and Prdx6.
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http://dx.doi.org/10.3390/antiox8090399DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6769615PMC
September 2019

Swelling Behavior of Polyacrylamide-Cellulose Nanocrystal Hydrogels: Swelling Kinetics, Temperature, and pH Effects.

Materials (Basel) 2019 Jun 28;12(13). Epub 2019 Jun 28.

Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, 100 Inha-Ro, Michuhol-Gu, Incheon 22212, Korea.

This paper reports swelling behavior of cellulose nanocrystal (CNC)-based polyacrylamide hydrogels prepared by a radical polymerization. The CNC acts as a nanofiller through the formation of complexation and intermolecular interaction. FTIR spectroscopy and XRD studies confirmed the formation of intermolecular bonds between the acrylamide and hydroxyl groups of CNC. The swelling ratio and water retention were studied in de-ionized (DI) water at room temperature, and the temperature effect on the swelling ratio was investigated. Further, the pH effect on the swelling ratio was studied with different temperature levels. Increasing the pH with temperature, the prepared hydrogel shows 6 times higher swelling ratio than the initial condition. The swelling kinetics of the developed hydrogels explains that the diffusion mechanism is Fickian diffusion mechanism. Since the developed hydrogels have good swelling behaviors with respect to pH and temperature, they can be used as smart materials in the field of controlled drug delivery applications.
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http://dx.doi.org/10.3390/ma12132080DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6650916PMC
June 2019

Excitatory neuron-specific SHP2-ERK signaling network regulates synaptic plasticity and memory.

Sci Signal 2019 03 5;12(571). Epub 2019 Mar 5.

Department of Physiology, Seoul National University College of Medicine, Seoul 03080, Korea.

Mutations in RAS signaling pathway components cause diverse neurodevelopmental disorders, collectively called RASopathies. Previous studies have suggested that dysregulation in RAS-extracellular signal-regulated kinase (ERK) activation is restricted to distinct cell types in different RASopathies. Some cases of Noonan syndrome (NS) are associated with gain-of-function mutations in the phosphatase SHP2 (encoded by ); however, SHP2 is abundant in multiple cell types, so it is unclear which cell type(s) contribute to NS phenotypes. Here, we found that expressing the NS-associated mutant SHP2 in excitatory, but not inhibitory, hippocampal neurons increased ERK signaling and impaired both long-term potentiation (LTP) and spatial memory in mice, although endogenous SHP2 was expressed in both neuronal types. Transcriptomic analyses revealed that the genes encoding SHP2-interacting proteins that are critical for ERK activation, such as GAB1 and GRB2, were enriched in excitatory neurons. Accordingly, expressing a dominant-negative mutant of GAB1, which reduced its interaction with SHP2, selectively in excitatory neurons, reversed SHP2-mediated deficits. Moreover, ectopic expression of GAB1 and GRB2 together with SHP2 in inhibitory neurons resulted in ERK activation. These results demonstrate that RAS-ERK signaling networks are notably different between excitatory and inhibitory neurons, accounting for the cell type-specific pathophysiology of NS and perhaps other RASopathies.
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http://dx.doi.org/10.1126/scisignal.aau5755DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6800025PMC
March 2019

Histone demethylase KDM3B regulates the transcriptional network of cell-cycle genes in hepatocarcinoma HepG2 cells.

Biochem Biophys Res Commun 2019 01 1;508(2):576-582. Epub 2018 Dec 1.

Department of Life Science, College of Natural Sciences, Chung-Ang University, 06974, Seoul, Republic of Korea. Electronic address:

Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third most lethal cancer worldwide. Although gene mutations associated with HCC development have been intensively studied, how epigenetic factors specifically modulate the functional properties of HCC by regulating target gene expression is unclear. Here we demonstrated the overexpression of KDM3B in liver tissue of HCC patients using public RNA-seq data. Ablation of KDM3B by CRISPR/Cas9 retarded the cell cycle and proliferation of hepatocarcinoma HepG2 cells. Approximately 30% of KDM3B knockout cells exhibited mitotic spindle multipolarity as a chromosome instability (CIN) phenotype. RNA-seq analysis of KDM3B knockout revealed significantly down-regulated expression of cell cycle related genes, especially cell proliferation factor CDC123. Furthermore, the expression level of Cyclin D1 was reduced in KDM3B knockout by proteosomal degradation without any change in the expression of CCND1, which encodes Cyclin D1. The results implicate KDM3B as a crucial epigenetic factor in cell cycle regulation that manipulates chromatin dynamics and transcription in HCC, and identifies a potential gene therapy target for effective treatment of HCC.
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http://dx.doi.org/10.1016/j.bbrc.2018.11.179DOI Listing
January 2019

Electroactive Hydrogels Made with Polyvinyl Alcohol/Cellulose Nanocrystals.

Materials (Basel) 2018 Sep 4;11(9). Epub 2018 Sep 4.

Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, 100 Inha-Ro, Nam-Gu, Incheon 22212, Korea.

This paper reports a nontoxic, soft and electroactive hydrogel made with polyvinyl alcohol (PVA) and cellulose nanocrystal (CNC). The CNC incorporating PVA-CNC hydrogels were prepared using a freeze⁻thaw technique with different CNC concentrations. Fourier transform infrared spectroscopy, thermogravimetric analysis, X-ray diffraction and scanning electron microscopy results proved the good miscibility of CNCs with PVA. The optical transparency, water uptake capacity and mechanical properties of the prepared hydrogels were investigated in this study. The CNC incorporating PVA-CNC hydrogels showed improved displacement output in the presence of an electric field and the displacement increased with an increase in the CNC concentration. The possible actuation mechanism was an electrostatic effect and the displacement improvement of the hydrogel associated with its enhanced dielectric properties and softness. Since the prepared PVA-CNC hydrogel is nontoxic and electroactive, it can be used for biomimetic soft robots, actively reconfigurable lenses and active drug-release applications.
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http://dx.doi.org/10.3390/ma11091615DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6163614PMC
September 2018

Structured illumination microscopy imaging reveals localization of replication protein A between chromosome lateral elements during mammalian meiosis.

Exp Mol Med 2018 08 28;50(8):1-12. Epub 2018 Aug 28.

Department of Life Sciences, Chung-Ang University, Seoul, 06974, Korea.

An important event enabling meiotic prophase I to proceed is the close juxtaposition of conjoined chromosome axes of homologs and their assembly via an array of transverse filaments and meiosis-specific axial elements into the synaptonemal complex (SC). During meiosis, recombination requires the establishment of a platform for recombinational interactions between the chromosome axes and their subsequent stabilization. This is essential for ensuring crossover recombination and proper segregation of homologous chromosomes. Thus, well-established SCs are essential for supporting these processes. The regulation of recombination intermediates on the chromosome axis/SC and dynamic positioning of double-strand breaks are not well understood. Here, using super-resolution microscopy (structured illumination microscopy), we determined the localization of the replication protein A (RPA) complex on the chromosome axes in the early phase of leptonema/zygonema and within the CEs of SC in the pachynema during meiotic prophase in mouse spermatocytes. RPA, which marks the intermediate steps of pairing and recombination, appears in large numbers and is positioned on the chromosome axes at the zygonema. In the pachynema, RPA foci are reduced but do not completely disappear; instead, they are placed between lateral elements. Our results reveal the precise structure of SC and localization dynamics of recombination intermediates on meiocyte chromosomes undergoing homolog pairing and meiotic recombination.
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http://dx.doi.org/10.1038/s12276-018-0139-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6113238PMC
August 2018

Deacetylase activity-independent transcriptional activation by HDAC2 during TPA-induced HL-60 cell differentiation.

PLoS One 2018 24;13(8):e0202935. Epub 2018 Aug 24.

Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul, Korea.

The human myeloid leukemia cell line HL-60 differentiate into monocytes following treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). However, the mechanism underlying the differentiation of these cells in response to TPA has not been fully elucidated. In this study, we performed ChIP-seq profiling of RNA Pol II, HDAC2, Acetyl H3 (AcH3), and H3K27me3 and analyzed differential chromatin state changes during TPA-induced differentiation of HL-60 cells. We focused on atypically active genes, which showed enhanced H3 acetylation despite increased HDAC2 recruitment. We found that HDAC2 positively regulates the expression of these genes in a histone deacetylase activity-independent manner. HDAC2 interacted with and recruited paired box 5 (PAX5) to the promoters of the target genes and regulated HL-60 cell differentiation by PAX5-mediated gene activation. Taken together, these data elucidated the specific-chromatin status during HL-60 cell differentiation following TPA exposure and suggested that HDAC2 can activate transcription of certain genes through interactions with PAX5 in a deacetylase activity-independent pathway.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0202935PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108480PMC
February 2019

Spindle pole body component 25 homolog expressed by ECM stiffening is required for lung cancer cell proliferation.

Biochem Biophys Res Commun 2018 06 2;500(4):937-943. Epub 2018 May 2.

Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul, 06974, Republic of Korea. Electronic address:

Accumulating evidence has shown that matrix stiffening in cancer tissue by the deposition of extracellular matrix (ECM) is closely related with severe tumor progression. However, much less is known about the genes affected by matrix stiffness and its signaling for cancer progression. In the current research, we investigated the differential gene expression of a non-small lung adenocarcinoma cell line, H1299, cultured under the conditions of soft (∼0.5 kPa) and stiff (∼40 kPa) matrices, mimicking the mechanical environments of normal and cancerous tissues, respectively. For integrated transcriptome analysis, the genes identified by ECM stiffening were compared with 8248 genes retrieved from The Cancer Genome Atlas Lung Adenocarcinoma (TCGA). In stiff matrix, 29 genes were significantly upregulated, while 75 genes were downregulated. The screening of hazard ratios for these genes using the Kaplan-Meier Plotter identified 8 genes most closely associated with cancer progression under the condition of matrix stiffening. Among these genes, spindle pole body component 25 homolog (SPC25) was one of the most up-regulated genes in stiff matrix and tumor tissue. Knockdown of SPC25 in H1299 cells using shRNA significantly inhibited cell proliferation with downregulation of the expression of checkpoint protein, Cyclin B1, under the condition of stiff matrix whereas the proliferation rate in soft matrix was not affected by SPC25 silencing. Thus, our findings provide novel key molecules for studying the relationship of extracellular matrix stiffening and cancer progression.
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http://dx.doi.org/10.1016/j.bbrc.2018.04.205DOI Listing
June 2018

Cellulose nanofibers isolated by TEMPO-oxidation and aqueous counter collision methods.

Carbohydr Polym 2018 Jul 7;191:65-70. Epub 2018 Mar 7.

Creative Research center for Nanocellulose Future Composite, Inha University, Incheon, South Korea. Electronic address:

In this research, cellulose nanofiber (CNF) was isolated by the combination of chemical 2,2,6,6-tetramethylpiperidine-1-oxylradical (TEMPO)-oxidation and physical aqueous counter collision (ACC) methods The combination of TEMPO-oxidation and ACC is an efficient method to isolate CNFs by reducing chemical usage in TEMPO-oxidation and saving energy in ACC along with controlling the size of CNFs. Two cellulose sources, hardwood bleached kraft pulp (HW) and softwood bleached kraft pulp (SW), were used for the CNF isolation with different TEMPO oxidation time and a defined number of ACC pass. The CNF properties were investigated and compared in term of morphology, crystallinity index, transparency and birefringence. The width of the isolated CNFs from HW is in the range of 15.1 nm-17.5 nm, and that of the SW CNFs is between 18.4 nm and 22 nm depending on the TEMPO oxidation time. This difference is due to the fact that SW is less oxidized than HW under the same chemical dosage, which results in larger width of SW-CNFs than HW-CNFs. The HW-CNF treated with TEMPO for over 2 h and isolated using ACC with 5 pass offers almost 90% transparency. Birefringence of CNFs exhibits that HW-CNFs show better birefringence phenomenon than SW-CNFs. The combination of TEMPO-oxidation and ACC methods is useful for isolating CNFs with its size control.
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http://dx.doi.org/10.1016/j.carbpol.2018.03.008DOI Listing
July 2018

Developmental biology and potential use of (Annelida: Hirudinea) as an "Evo-Devo" model organism.

Front Zool 2017 28;14:60. Epub 2017 Dec 28.

School of Biological Sciences, College of Natural Sciences, Chungbuk National University, Cheongju, Chungbuk 28644 Republic of Korea.

Background: The need for the adaptation of species of annelids as "Evo-Devo" model organisms of the superphylum Lophotrochozoa to refine the understanding of the phylogenetic relationships between bilaterian organisms, has promoted an increase in the studies dealing with embryonic development among related species such as leeches from the Glossiphoniidae family. The present study aims to describe the embryogenesis of (Oka, 1910), a freshwater glossiphoniid leech, chiefly distributed in East Asia, and validate standard molecular biology techniques to support the use of this species as an additional model for "Evo-Devo" studies.

Results: undergoes direct development, and follows the highly conserved clitellate annelid mode of spiral cleavage development; the duration from the egg laying to the juvenile stage is ~7.5 days, and it is iteroparous, indicating that it feeds and deposits eggs again after the first round of brooding, as described in several other glossiphoniid leech species studied to date. The embryos hatch only after complete organ development and proboscis retraction, which has not yet been observed in other glossiphoniid genera. The phylogenetic position of within the Glossiphoniidae family has been confirmed using cytochrome c oxidase subunit 1 (CO1) sequencing. Lineage tracer injections confirmed the fates of the presumptive meso- and ectodermal precursors, and immunostaining showed the formation of the ventral nerve system during later stages of development. Further, the spatiotemporal expression of an EF-hand calcium-binding protein Calsensin ortholog was characterized, which showed a specific pattern in both the ventral and peripheral nervous systems during the later stages.

Conclusions: Our description of the embryonic development of under laboratory conditions provides new data for further comparative studies with other leech and lophotrochozoa model organisms. Moreover, it offers a basis for the establishment of this species as a model for future "Evo-Devo" studies.
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http://dx.doi.org/10.1186/s12983-017-0240-yDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5745604PMC
December 2017

Cellulose long fibers fabricated from cellulose nanofibers and its strong and tough characteristics.

Sci Rep 2017 12 15;7(1):17683. Epub 2017 Dec 15.

Creative Research Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, Incheon, 222212, Republic of Korea.

Cellulose nanofiber (CNF) with high crystallinity has great mechanical stiffness and strength. However, its length is too short to be used for fibers of environmentally friendly structural composites. This paper presents a fabrication process of cellulose long fiber from CNF suspension by spinning, stretching and drying. Isolation of CNF from the hardwood pulp is done by using (2, 2, 6, 6-tetramethylpiperidine-1-yl) oxidanyl (TEMPO) oxidation. The effect of spinning speed and stretching ratio on mechanical properties of the fabricated fibers are investigated. The modulus of the fabricated fibers increases with the spinning speed as well as the stretching ratio because of the orientation of CNFs. The fabricated long fiber exhibits the maximum tensile modulus of 23.9 GPa with the maximum tensile strength of 383.3 MPa. Moreover, the fabricated long fiber exhibits high strain at break, which indicates high toughness. The results indicate that strong and tough cellulose long fiber can be produced by using ionic crosslinking, controlling spinning speed, stretching and drying.
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http://dx.doi.org/10.1038/s41598-017-17713-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5732198PMC
December 2017

Transcriptome analysis for UVB-induced phototoxicity in mouse retina.

Environ Toxicol 2018 Jan 17;33(1):52-62. Epub 2017 Oct 17.

Department of Life Science, College of Natural Sciences, Chung-Ang University, Seoul, 06974, Republic of Korea.

Throughout life, the human eye is continuously exposed to sunlight and artificial lighting. Ambient light exposure can lead to visual impairment and transient or permanent blindness. To mimic benign light stress conditions, Mus musculus eyes were exposed to low-energy UVB radiation, ensuring no severe morphological changes in the retinal structure post-exposure. We performed RNA-seq analysis to reveal the early transcriptional changes and key molecular pathways involved before the activation of the canonical cell death pathway. RNA-seq analysis identified 537 genes that were differentially modulated, out of which 126 were clearly up regulated (>2-fold, P < .01) and 51 were significantly down regulated (<2-fold, P < .01) in response to UVB irradiation in the mouse retina. Gene ontology analysis revealed that UVB exposure affected pathways for cellular stress and signaling (eg, Creb3, Ddrgk1, Grin1, Map7, Uqcc2, Uqcrb), regulation of chromatin and gene expression (eg, Chd5, Jarid2, Kat6a, Smarcc2, Sumo1, Zfp84), transcription factors (eg, Asxl2, Atf7, Per1, Phox2a, Rxra), RNA processing, and neuronal genes (eg, B4gal2, Drd1, Grm5, Rnf40, Rnps1, Usp39, Wbp4). The differentially expressed genes from the RNA-seq analysis were validated by quantitative PCR. Both analyses yielded similar gene expression patterns. The genes and pathways identified here improve the understanding of early transcriptional responses to UVB irradiation. They may also help in elucidating the genes responsible for the inherent susceptibility of humans to UVB-induced retinal diseases.
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http://dx.doi.org/10.1002/tox.22494DOI Listing
January 2018

Performance improvement of miniaturized ZnO nanowire accelerometer fabricated by refresh hydrothermal synthesis.

R Soc Open Sci 2017 Sep 13;4(9):170557. Epub 2017 Sep 13.

Creative Research Center for Nanocellulose Future Composites, Department of Mechanical Engineering, Inha University, 100 Inha-Ro, Nam-Gu, Incheon 22212, Korea.

Miniaturized accelerometers are necessary for evaluating the performance of small devices, such as haptics, robotics and simulators. In this study, we fabricated miniaturized accelerometers using well-aligned ZnO nanowires. The layer of ZnO nanowires is used for active piezoelectric layer of the accelerometer, and copper was chosen as a head mass. Seedless and refresh hydrothermal synthesis methods were conducted to grow ZnO nanowires on the copper substrate and the effect of ZnO nanowire length on the accelerometer performance was investigated. The refresh hydrothermal synthesis exhibits longer ZnO nanowires, 12 µm, than the seedless hydrothermal synthesis, 6 µm. Performance of the fabricated accelerometers was verified by comparing with a commercial accelerometer. The sensitivity of the fabricated accelerometer by the refresh hydrothermal synthesis is shown to be 37.7 pA g, which is about 30 times larger than the previous result.
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http://dx.doi.org/10.1098/rsos.170557DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627100PMC
September 2017

Regulation of Noncoding Transcriptome in Developing Photoreceptors by Rod Differentiation Factor NRL.

Invest Ophthalmol Vis Sci 2017 09;58(11):4422-4435

Neurobiology-Neurodegeneration and Repair Laboratory, National Eye Institute, National Institutes of Health, Bethesda, Maryland, United States.

Purpose: Transcriptome analysis by next generation sequencing allows qualitative and quantitative profiling of expression patterns associated with development and disease. However, most transcribed sequences do not encode proteins, and little is known about the functional relevance of noncoding (nc) transcriptome in neuronal subtypes. The goal of this study was to perform a comprehensive analysis of long noncoding (lncRNAs) and antisense (asRNAs) RNAs expressed in mouse retinal photoreceptors.

Methods: Transcriptomic profiles were generated at six developmental time points from flow-sorted Nrlp-GFP (rods) and Nrlp-GFP;Nrl-/- (S-cone like) mouse photoreceptors. Bioinformatic analysis was performed to identify novel noncoding transcripts and assess their regulation by rod differentiation factor neural retina leucine zipper (NRL). In situ hybridization (ISH) was used for validation and cellular localization.

Results: NcRNA profiles demonstrated dynamic yet specific expression signature and coexpression clusters during rod development. In addition to currently annotated 586 lncRNAs and 454 asRNAs, we identified 1037 lncRNAs and 243 asRNAs by de novo assembly. Of these, 119 lncRNAs showed altered expression in the absence of NRL and included NRL binding sites in their promoter/enhancer regions. ISH studies validated the expression of 24 lncRNAs (including 12 previously unannotated) and 4 asRNAs in photoreceptors. Coexpression analysis demonstrated 63 functional modules and 209 significant antisense-gene correlations, allowing us to predict possible role of these lncRNAs in rods.

Conclusions: Our studies reveal coregulation of coding and noncoding transcripts in rod photoreceptors by NRL and establish the framework for deciphering the function of ncRNAs during retinal development.
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http://dx.doi.org/10.1167/iovs.17-21805DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5584472PMC
September 2017
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