Deubiquitinase YOD1 potentiates YAP/TAZ activities through enhancing ITCH stability

Kim, Youngeun,Kim, Wantae,Song, Yonghee,Kim, Jeong-Rae,Cho, Kyungjoo,Moon, Hyuk,Ro, Simon Weonsang,Seo, Eunjeong,Ryu, Yeon-Mi,Myung, Seung-Jae,Jho, Eek-Hoon National Academy of Sciences 2017 Proceedings of the National Academy of Sciences Vol.114 No.18

<P>Hippo signaling controls the expression of genes regulating cell proliferation and survival and organ size. The regulation of core components in the Hippo pathway by phosphorylation has been extensively investigated, but the roles of ubiquitination-deubiquitination processes are largely unknown. To identify deubiquitinase(s) that regulates Hippo signaling, we performed unbiased siRNA screening and found that YOD1 controls biological responses mediated by YAP/TAZ. Mechanistically, YOD1 deubiquitinates ITCH, an E3 ligase of LATS, and enhances the stability of ITCH, which leads to reduced levels of LATS and a subsequent increase in the YAP/TAZ level. Furthermore, we show that the miR-21-mediated regulation of YOD1 is responsible for the cell-density-dependent changes in YAP/TAZ levels. Using a transgenic mouse model, we demonstrate that the inducible expression of YOD1 enhances the proliferation of hepatocytes and leads to hepatomegaly in a YAP/TAZ-activity-dependent manner. Moreover, we find a strong correlation between YOD1 and YAP expression in liver cancer patients. Overall, our data strongly suggest that YOD1 is a regulator of the Hippo pathway and would be a therapeutic target to treat liver cancer.</P>

Assessment of foodservice quality and identification of improvement strategies using hospital foodservice quality model

Kyungjoo Kim,Minyoung Kim,Kyung-Eun Lee 한국영양학회 2010 Nutrition Research and Practice Vol.4 No.2

The purposes of this study were to assess hospital foodservice quality and to identify causes of quality problems and improvement strategies. Based on the review of literature, hospital foodservice quality was defined and the Hospital Foodservice Quality model was presented. The study was conducted in two steps. In Step 1, nutritional standards specified on diet manuals and nutrients of planned menus, served meals, and consumed meals for regular, diabetic, and low-sodium diets were assessed in three general hospitals. Quality problems were found in all three hospitals since patients consumed less than their nutritional requirements. Considering the effects of four gaps in the Hospital Foodservice Quality model, Gaps 3 and 4 were selected as critical control points (CCPs) for hospital foodservice quality management. In Step 2, the causes of the gaps and improvement strategies at CCPs were labeled as “quality hazards” and “corrective actions”, respectively and were identified using a case study. At Gap 3, inaccurate forecasting and a lack of control during production were identified as quality hazards and corrective actions proposed were establishing an accurate forecasting system, improving standardized recipes, emphasizing the use of standardized recipes, and conducting employee training. At Gap 4, quality hazards were menus of low preferences, inconsistency of menu quality, a lack of menu variety, improper food temperatures, and patients’ lack of understanding of their nutritional requirements. To reduce Gap 4, the dietary departments should conduct patient surveys on menu preferences on a regular basis, develop new menus, especially for therapeutic diets, maintain food temperatures during distribution, provide more choices, conduct meal rounds, and provide nutrition education and counseling. The Hospital Foodservice Quality Model was a useful tool for identifying causes of the foodservice quality problems and improvement strategies from a holistic point of view.

Establishment of 3D Multicellular Tumor Spheroids (MCTS) using Hepatocellular Carcinoma and Stromal cells for Screening Anti-cancer Therapeutic Agents

( Kyungjoo Cho ),( Hyuk Moon ),( Soonyoung Shin ),( Simon W. Ro ),( Hye Won Lee ),( Beom Kyung Kim ),( Do Young Kim ),( Kwang-hyub Han ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer in adults and the second leading cause of cancer-related deaths worldwide. Tumor microenvironment is composed of myofibroblasts, fibroblasts, immune-inflammatory cells, extracellular matrix, blood vessels, etc. and closely involved in multiple facets of tumorigenesis. Studies have shown that response to chemotherapy is highly affected by drug penetration through tumor tissue, highlighting the role of tumor microenvironment in cancer chemotherapy. Compared with tumor cell monolayer culture, multi-cellular tumor spheroid (MCTS) is superior in mimicking tumor microenvironment and thus a suitable model for studying drug penetration into tumor. The purpose of this study is to establish the MCTS model to investigate the interaction with the microenvironment in tumor and to investigate the effect of microenvironment on drug permeability. Methods: To generate multicellular tumor spheroids, HCC cells were seeded at a density of 6 × 103 cells/well in 96-well round-bottom ultra-low attachment microplates. The plates were incubated for 3 days at 37 °C in a humidified atmosphere of 5% CO2. To generate MCTS, HCC cells and stromal cells (LX2, WI38, and HUVECs) were mixed at a 1:1 ratio. Diameter was measured using a confocal microscope and an image analyzer to determine the compactness of spheroid. Protein expression levels in MCTS were determined by immunoblots. For drug penetration study, fluorescent chemicals (e.g., verteporfin) were used and the distribution of drug within MCTS was determined by a LSM700 confocal microscope with a 425 to 440nm excitation and a 700 to 730nm emission filter set. Results: Various multi-cellular tumor spheroid (MCTS) models were developed using HCC cell lines with various degrees of differentiation such as SNU449 (Well differentiated), SNU3059 and SNU3160 (moderately differentiated), and Hep3B (poorly differentiated). The volume, shape, and compactness of HCC MCTS were heterogeneous depending on the differentiation degree. Well differentiated SNU449 MCTS showed the least compactness of tumor spheroids, while Hep3B MCTS had the highest compactness. Of note, YAP/TAZ levels in HCC MCTS were significantly different. SNU449 MCTS model had a low level of YAP/TAZ, while Hep3B MCTS model had the highest level of YAP/TAZ expression. Drug penetration into tumor spheroids was significantly retarded due to the multi-cellular components within HCC MCTS. HCC MCTS with higher YAP/TAZ levels increased the compactness and inhibited drug penetration. Conclusions: In this study, diverse MCTS models have been developed using HCC of different degrees of differentiation and stromal cells such as HSCs, fibroblasts, and endothelial cells. MCTS with poorly differentiated HCC showed an increased compactness of spheroids, an elevated level of YAP/TAZ and a limited drug penetration. Reducing tumor compactness or stromal activation should be considered to improve a response to chemotherapy in patients with advanced HCC.

Media Politics in Japan and Korea-Japan Relations

KIM KYUNGJOO 한양대학교 일본학국제비교연구소 2008 비교일본학 Vol.18 No.-

T58A Mutation Does Not Enhance Tumorigenic Potentials of C-MYC in the Liver

( Kyungjoo Cho ),( Sook In Chung ),( Hyuk Moon ),( Daeyoung Kim ),( Do Young Kim ),( Simon W. Ro ),( Kwang-hyub Han ) 대한간학회 2017 춘·추계 학술대회 (KASL) Vol.2017 No.1

Aims: Liver cancer is a major health concern worldwide, ranking the second among cancer-related mortality. The c-Myc gene is epigenetically altered in almost 50% of human liver cancers, leading to persistent over-expression of c-Myc. Mutation at codon 58 of c-Myc (c-Myc^T58A) can enhance oncogenic potentials of c-Myc through suppressing apoptotic signaling cascades or stabilizing the oncoprotein. In this study, we compared tumorigenic potentials between c-Myc^T58A and the wild-type (WT) c-Myc in the liver. Methods: Transgenic mouse models expressing c-Myc^T58A and WT c-Myc were developed using hydrodynamic transfection. Transposons encoding an activated from of human H-RAS were mixed with transposons encoding either c-Myc^T58A or WT c-Myc. The DNA mixtures were injected into the lateral tail veins of 6-week-old C57BL/6 mice. Mice were monitored at least twice per week and sacrificed when moribund. Tumor-bearing livers were formalin fixed for hematoxylin- eosin staining and immunohistochemistry. Results: Hepatocellular carcinomas (HCC) were induced by co- expression of HRAS with either c-Myc^T58A or WT c-Myc with 100% penetration. There was no significant difference in animal survivals between the c-Myc^T58A and WT c-Myc groups. The numbers and sizes of tumors were similar between the two groups. Cellular proliferation (determined by Ki-67 staining) and apoptosis levels (by TUNEL assay) were also similar between c-Myc^T58A and WT c-Myc groups. Finally, there was no difference in phenotypes of malignant hepatocytes between the two groups. Conclusions: T58A mutation does not enhance tumorigenic potentials of c-MYC in our transgenic mouse models. No downregulation of apoptosis was detected in c-Myc^T58A, compared with WT c-Myc.

납, 카드뮴, 니켈 공존 하에서의 자성중심 덴드리머의 경쟁 흡착에 관한 연구

김경주(Kyungjoo Kim),박재우(Jaewoo Park) 한국지반환경공학회 2015 한국지반환경공학회 학술발표회논문집 Vol.2015 No.09

Kinase Suppressor of Ras 1 Promotes YAP/TAZ-Mediated Tumorigenesis in the Liver

( Hyuk Moon ),( Kyungjoo Cho ),( Soonyoung Shin ),( Simon W. Ro ),( Beom Kyung Kim ),( Seung Up Kim ),( Jun Yong Park ),( Sang Hoon Ahn ),( Do Young Kim ),( Kwang-hyub Han ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Liver cancer is the second most common cause of cancer-related deaths worldwide. The Hippo signaling pathway is tumor suppressive, as its inactivation leads to tissue overgrowth and tumor formation via YAP- or TAZ-mediated transcriptional activation. YAP is overexpressed in 62% of patients with hepatocellular carcinoma (HCC) and in almost 90% of human cholangiocarcinoma (CCC). Kinase Suppressor of Ras 1 (KSR1) is a scaffold protein for the Ras/Raf/MEK/ERK signaling pathway, promoting activation of MEK and ERK. Pro-tumorigenic roles of KSR1 in Ras-activated cancers have been verified in murine models for lung, skin, and pancreatic cancers. In this study, we have investigated the role of KSR1 in YAP/TAZ-mediated hepatocarcinogenesis. Methods: Transposons were constructed encoding KSR1 and an activated from of TAZ (TAZ^S89A). Transposons were hydrodynamically delivered to livers of 6-week-old C57BL/6 mice. Mice were monitored at least twice per week and sacrificed when moribund. Tumor-bearing livers were formalin fixed for hematoxylin-eosin staining and immunohistochemistry. Results: Analysis of gene expression levels in human HCC and CCC samples deposited in The Cancer Genome Atlas (TCGA) revealed that KSR1 was significantly upregulated both in human HCC and CCC, compared with non-tumoral surrounding livers (P<0.0001 in HCC and P=0.0063 in CCC). Co-expression of KSR1 and an activated form of TAZ (TAZ^S89A) led to the development of both HCC and CCC in the murine livers, while expression of TAZ^S89A alone failed to induce hepatic tumors. Conclusions: KSR1 promotes hepatocarcinogenesis, both in HCC and CCC. Suppression of KSR1 might be an attractive therapeutic option for both types of hepatic malignancies.

Inhibition of Dickkopf-1 Enhances Antitumor Efficacy of Sorafenib in Hepatocellular Carcinoma

( Sanghyun Seo ),( Kyungjoo Cho ),( Hye Jung Park ),( Hye Won Lee ),( Beom Kyung Kim ),( Jun Yong Park ),( Do Young Kim ),( Sang Hoon Ahn ),( Seung Up Kim ) 대한간학회 2021 춘·추계 학술대회 (KASL) Vol.2021 No.1

F-122 Is Inhaled corticosteroid associated with less exacerbation in subjects with mild to moderate airflow limitation?

( Soo Jung Kim ),( Jin Hwa Lee ),( Yon Ju Ryu ),( Jung Hyun Chang ),( Chin Kook Rhee ),( Kyungjoo Kim ),( Kwang Ha Yoo ),( Ki-suck Jung ) 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.-

Background: It has been unclear whether inhaled corticosteroids (ICS) clinically benefit patients with early-stage of COPD. Nevertheless, in the real world, ICS have been prescribed to patients with FEV1 ≥60% predicted. The objective of this study was to evaluate whether ICS was associated with reduced exacerbations in patients with FEV1 ≥60% predicted. Methods: A retrospective cohort was constructed from the combination of data from both the Korean National Health and Nutrition Examination Survey and the national health insurance reimbursement databases during 2007-2012. Patients aged ≥20 years with a pre-bronchodilator FEV1 ≥60% predicted and FEV1/FVC <0.7 were included. Acute exacerbations requiring hospitalization, intensive care unit (ICU) admission, and emergency room (ER) visits were evaluated. Results: Among 365 patients with FEV1 ≥60% predicted, ICS were prescribed for 88 patients (24%). ICS users showed lower FEV1 and higher household income compared to non-users. Adjusted for age, sex, BMI, FEV1, and household income, use of ICS was not associated with decreased risk of acute exacerbation. However, ICU use was associated with increased risk of hospitalization due to pneumonia (adjusted odds ratio, 2.86; 95% CI 1.39 to 6.04), but neither ICU admission nor ER visits. Additionally, ICS use was not associated with increased medical cost. Conclusions: In patients with FEV1 ≥60% predicted, ICS may not reduce acute exacerbation, but increase the risk of hospitalization due to pneumonia. Therefore, prescribing ICS to these patients requires careful consideration.

Autophagy Inhibitor, Chloroquine Did Not Affect Tumor Development in a Transgenic Mouse Model of HCC

( Soonyoung Shin ),( Hyuk Moon ),( Kyungjoo Cho ),( Simon W. Ro ),( Beom Kyung Kim ),( Seung Up Kim ),( Do Young Kim ),( Kwang-hyub Han ) 대한간학회 2018 춘·추계 학술대회 (KASL) Vol.2018 No.1

Aims: Autophagy is an intracellular recycling process by which damaged or superfluous proteins are delivered to lysosomes for degradation, and then utilized as energy resources and macromolecular precursors. Autophagy in cancer is a highly debated subject. Research has shown that autophagy can become either tumor-promoting or tumor-suppressive depending on cellular or genetic context. Here, we investigated the role of autophagy in hepatocellular carcinoma (HCC) by applying an autophagy inhibitor, chloroquine to a transgenic mouse model of HCC. Methods: Transposons were constructed encoding an activated from of RAS (HRASG12V) and short hairpin suppressing P53 (shp53). Transposons were hydrodynamically delivered to livers of 6-week-old C57BL/6 mice. Mice were administered intraperitoneally with chloroquine at a daily dose of 60mg/kg for five weeks. Control mice were given a phosphate buffered saline (PBS). Mice were monitored at least twice per week. Results: The sizes and numbers of tumor nodules were similar between chloroquine group and control when livers were harvested at 5 weeks after the delivery of oncogenes. Animal survivals were not significantly different between the two groups, suggesting that the treatment with chloroquine does not affect liver tumorigenesis induced by HRASG12V plus shp53. Conclusions: Our study suggests that autophagy inhibition has a minimal role in HCC under the genetic context of RAS signaling activation and P53 downregulation.