CORO7에 의한 Hippo 신호전달체계 조절에 관한 연구

학위논문 (석사)-- 서울대학교 대학원 : 자연과학대학 생명과학부, 2018. 8. 정종경.From Drosophila to mammals, the Hippo pathway plays crucial roles in control of organ growth and tissue homeostasis. Through rigorous studies for the last two decades, the core kinase cassette comprised of two kinases (MST1/2 and LATS1/2) and two scaffolds (SAV1 and MOB1) has been discovered to be essential for the regulation of Hippo signaling. However, how and where its formation and activation happens are not fully understood. Here, I identify that CORO7 positively regulates the Hippo pathway through the analysis of protein interaction database and genetic screening in Drosophila. I demonstrate that CORO7 is necessary for Hippo signaling activation induced by serum deprivation, contact inhibition and cytoskeleton damage. I also show that CORO7 physically interacts with Hippo core components and functions as a scaffold for the core kinase cassette. Interaction of CORO7 with SAV1 and MST1/2 is regulated by Hippo activating signals, suggesting a multi-step binding regulation of Hippo core kinase cassette by CORO7. My results provide molecular insights of Hippo signaling regulation and have implications for understanding Hippo-related diseases.INTRODUCTION 1 RESULTS 4 CORO7 is a positive regulator of the activation of the Hippo pathway in mammalian cells 4 Physical interaction of CORO7 with Hippo pathway components 10 CORO7 is important for the formation of the Hippo core kinase cassette 19 Regulation of CORO7 interaction with Hippo components by Hippo activating signals 20 SAV1 binds to NF2 instead of CORO7 upon the Hippo pathway activation 23 DISCUSSION 26 MATERIALS and METHODS 31 REFERENCES 35 국문 초록 41Maste

Three Cases of Gastroesophageal Reflux Disease Treated by Laparoscopic Fundoplication

Medical treatment so far takes the major portion in the treatment of gastroesophageal reflux disease (GERD). Histamine-2 receptor antagonists, proton pump inhibitors and antacids, which are used in the medical treatment of GERD, decreases intragastric acidity, and therefore decreases acid reflux. However, recurrences are frequently observed after the cessation of medication. On the other hand, fundoplication, the surgical management of GERD, aims for physiologic reconstruction of esophagogastric junction, and for correcting the pathogenetic cause of GERD. But fundoplication is linked to risks related to surgery and general anesthesia. Laparoscopic funcoplication minimizes these risks. We recently experienced three cases of GERD improved by laparoscopic 270o fundoplication. One patient was intolerant of long-term proton pump inhibitor treatment, and two patients had hiatal hernia, and one of them showed failure to medical treatment. This report describes these three cases with a brief review of literatures survey.ope

Implication of IGFBP-3 Promoter Polymorphism in Type 2 DM in Elderly

Background: Insulin-like growth factor (IGF) binding protein-3 (IGFBP-3) is a major determinant of circulating levels of the IGFs and is clinically useful for the evaluation of GH deficiency and for predicting the response to GH treatment. Circulating levels of IGF-1 and IGFBP-3 vary considerably among individuals and have been implicated in various medical conditions. We investigated the IGFBP-3 genotype distribution in Korean elderly and relations between IGFBP-3 promoter -202 locus A/C polymorphism and medical conditions. Methods: Single nucleotide polymorphism at the -202 locus of the IGFBP-3 promoter was genotyped for 100 samples from a study population who is older than 60 years. Results: The frequency of each polymorphic variation at the -202 locus in these study population was AA=43 (43%), AC=48 (48%), and CC=8 (8%). In both genders, the subjects with the CC genotype were taller, whereas those with AC genotype had higher weight, BMI and BSA comparing those with the other genotypes, but these facts did not reach statistical significance. The prevalence of type 2 DM and the mean fasting blood glucose level did not differ among genotypes in the female gender. In the male population, however, the prevalence of type 2 DM was higher in the subjects with the AC genotype (P=0.016) which is reflected by the elevated mean fasting glucose level (P=0.106). Conclusion: IGFBP-3 promoter -202 locus A/C polymorphism, which reflects its serum level and further implicates IGFs bioactivity, was associated with a higher prevalence of type 2 DM in the male gender, suggesting IGFBP-3 might play a role in blood glucose homeostasis.ope

The Frequency of Anti-GAD Antibody in Non-obese, Adult-onset Type 2 Diabetes in Korea and Clinical and Biological Characteristics According to Anti-GAD Antibody

Background: Some of the characteristic features of diabetes mellitus in Korea are that 70-80% of patients are non-obese or adult-onset type, and type 1 diabetes is very rare. Occasionally, autoantibodies to glutamic acid decarboxylase(GAD) are found in typical, type 2 diabetes mellitus patients(T2DM). The role of the autoantibody to GAD in T2DM is unknown. The aim of this study was to determine the clinical and biochemical characteristics between GAD-positive and GAD-negative non-obese, adult-onset diabetics in Korea. Methods: A cohort of 428 type 2 diabetes patients was included. The measured autoantibodies to GAD were measured, and the C-peptide and HbA1c levels, anthropometric data(weight, height, body mass index and waist circumference), blood pressure and lipid profiles compared between the two groups. Results: Compared to the antibody-negative group(n=374; 87.4%), patients with the anti-GAD antibody(n=54; 12.6%) had significantly lower C-peptide levels and were significantly younger. The anti GAD-positive group had a lower BMI, more frequently needed insulin supplements, and a lower prevalence of hypertension. There were no significant differences in gender and family history of diabetes between the two groups. Conclusion: The presence of the autoantibody to GAD allowed the group with more deteriorated beta-cell function and more frequent need for insulin supplements, but a lower prevalence of obesity and hypertension to be determined.ope

Apolipoprotein E and ACE genetic polymorphism and nephropathy in type 2 diabetic patients

Background : The aim of this study was to investigate the association between apo E and ACE genetic polymorphism and diabetic nephropathy. Methods : One hundred eighteen patients with type 2 diabetes who had a duration of diabetes longer than 8 years were divided into the three apo E groups (E2, E3, E4) and three ACE groups (II, ID, DD). Plasma levels of lipids were measured. The frequency of diabetic nephropathy and clinical and biochemical characteristics were compared among the Apo E and ACE genotype groups. Results : The frequency of overt nephropathy was significantly greater in apo E2 patients with diabetes (46.7%) than apo E3 (16.7%) or apo E4 patients (10.5%). Logistical regression analysis showed that odds ratio of apo E2 and apo E4 genotypes for the presence of overt nephropathy were 4.779 (p<0.01) and 0.643 (p=0.583), respectively. Plasma TG levels were significantly greater in apo E2 patients. This study did not show an association between ACE gene polymorphism and diabetic nephropathy, and no interaction between Apo E and ACE gene polymorphism. Conclusion : Apo E2 is a prognostic risk factor for diabetic nephropathy in Korean type 2 diabetes. TG may have an important role of diabetic nephropathy. There were not synergistic effect between Apo E and ACE gene polymorphism in diabetic nephropathyope

Role of Activation of NF- B and AP-1 by Oxidative Stress in Atherosclerosis in Diabetic Patients

Background: The aim of this study was to evaluate the possible role of NF- B activation and AP-1 by oxidative stress in atherosclerosis in diabetic patients by measuring the carotid intima-media thickness, intracellular ROS generation and activation of transcription factors, including nuclear factor-kappa B (NF- B) and activator protein-1 (AP-1). Methods: Sixty-six patients (28 males, 38 females; age 56.1 13.4 years; duration of diabetes 115.7 83.4 months) with type 2 diabetes mellitus (DM) were selected for this study. The DM patients included in this study were divided into those with a normal carotid intima-media thickness (Group II) and those with an increased intima-media thickness (Group III). 57 healthy controls matched for age and sex with the DM patients (Group I) were randomly selected. Dichlorodifluorescein (DCF)-sensitive intracellular ROS was measured by fluorescent spectrometry. The activities of NF- B and AP-1 in PBMCs were measured by an electrophoretic mobility shift assay. Results: No differences were evident between the groups in terms of gender, age, BMI, blood pressure, total cholesterol, triglyceride, LDL-cholesterol and HDL-cholesterol. Spontaneous and H2O2 (or phorbol-12-myristate-13-acetate, PMA) stimulated ROS were significantly higher in the PBMCs from the DM patients with an increased intima-media thickness (Group III) than in those without (Group II), and were also higher in the control group (Group I). Moreover, the activities of NF- B and AP-1 were significantly higher in Group III than in Groups I or II. Conclusion: The present study demonstrates that intracellular ROS generation, and NF- B and AP-1 activation in PBMCs strongly correlates with the carotid artery IMT. These clinical results suggest that increased oxidative stress in PBMCs may play a role in the pathogenesis of atherosclerosis in DM patientsope

Risk Factors for Asymptomatic Atherosclerosis in Elderly Diabetic Patients without Diabetic Microvascular Complications

Background: Diabetes Mellitus is a major independent risk factor for atherosclerosis. We have no information on the prevalence of asymptomatic atherosclerosis in diabetic patients before the onset of microvascular diseases. The aims of this study was to evaluate risk factors for the atherosclerosis in elderly type 2 diabetic patients without the microvascular diseases. Methods: We evaluated atherosclerotic change of carotid arteries in 60 elderly Korean type 2 diabetic patients who had neither atherosclerotic vascular diseases nor diabetic microvascular complications.When atherosclerotic change was defined as the mean intima-media thickness (IMT) of ＞1.0 mm and/or the presence of plaque lesion, 28.3% of patients had atherosclerosis of the carotid arteries. Results: Risk factors for the carotid atherosclerosis were age, low-density lipoprotein (LDL)-cholesterol, hypertension, and diabetes treatment. Age and LDL-cholesterol were associated with mean IMT. Fasting plasma glucose, glycosylated hemoglobin (HbA1c), and known diabetes duration remained unassociated with any parameters of asymptomatic atherosclerosis of the carotid arteries. Conclusion: Glycemic control is unrelated with asymptomatic atherosclerosis in elderly type 2 diabetic patients without diabetic microvascular complications. Conventional risk factors and diabetes treatment are independently associated with atherosclerosis of the carotid arteries in these patients.ope

The Degree of Atherosclerosis and the Metabolic Characteristics according to the Abdominal Obesity in Type 2 Diabetic Patients

Background: Many of the maturity-onset type 2 diabetic patients with hypertension and dyslipidemia in Korea are not associated with obesity. However, these patients are at risk for developing macrovascular complications such as atherosclerosis due to hyperinsulinemia, insulin resistance and abdominal obesity. The aims of this study were to compare the clinical and biochemical differences between the type 2 diabetic patients that are with and without abdominal obesity, and we also wished to investigate the degree of insulin resistance and atherosclerosis in these patients. Methods: Among 530 type 2 diabetes mellitus (DM) patients, the percentages of under-weight (UW), normal-weight (NW), over-weight (OW) and obese (OB) (BMI or =30, respectively) subjects were 8.9%, 62.1%, 25.1% and 3.9%, respectively. To evaluate the severity of their atherosclerosis, the coronary artery calcification (CAC) score was measured by electron beam computed tomography, and the intima-media thickness (IMT) of the common carotid artery and the ankle-brachial pressure index (ABPI) were also measured. The Insulin sensitivity index (ISI) was measured by the plasma glucose disappearance rate (kitt: %/min). Results: 1. There were no differences in age, duration of DM and the HbA1c levels according to BMI for both the men and women, but the waist-hip ratio (WHR) and systolic blood pressure (SBP) were significantly different among each group. Serum triglyceride (TG), HDL-cholesterol (HDL-C), free fatty acid (FFA), fibrinogen, and fasting c-peptide levels, {excluding total cholesterol (TC)}, were also significantly different. The ISI, which is a marker for insulin resistance, as well correlated with the patients’ BMI. Subjects having an with ISI above 2.5%/min were considered as having insulin resistance, and 28%, 60%, 68% and 75% of patients in the UW, NW, OW and OB groups, respectively, demonstrated insulin resistance. The visceral fat area/subcutaneous fat area ratio and visceral fat area/thigh muscle area ratio also increased with BMI. 2. The median values of the WHR were 0.95 for the men and 0.91 for the women. There were no significant differences for age, BMI, duration of DM and HbA1c between patients with and without abdominal obesity, but the SBP, TG, HDL-C, FFA, fibrinogen and ISI were significantly different between those two groups. 3. For the OW group as well as the NW group, the carotid IMT, ABPI and CAC scores were significantly different between the patients with and without abdominal obesity. However, there were no differences between the NW group and the OW group. Conclusion: In conclusion, those patients with abdominal obesity, regardless of their BMIs, have a higher prevalence for atherosclerosis, dyslipidemia, and hypertension, compared to those patients without abdominal obesity. Therefore, it is important to screen for atherosclerosis and to manage it accordingly, for the patients with insulin resistance or abdominal obesity in order to decrease their risk of developing atherosclerotic events.ope

Relationship of LDL Particle Size to IMT and Insulin Resistance in Non-Diabetic Adult

Background: The aims of this study were to investigate the predictor of the low density lipoprotein(LDL) particle size and the relationship of the LDL particle size to the levels of insulin resistance and the carotid intima-media thickness (IMT) in healthy Koreans. Methods: The subjects were 47 and 89 clinically healthy males and females, aged between 32 and 70years, without medications that could potentially alter glucose and lipid metabolisms. The mean LDL particle size was determined by polyacrylamide tube gel electrophoresis(LipoprintⓇ LDL, Quantimetrix), the insulin resistance using a short insulin tolerance test kit, and the subclinical atherosclerosis from the carotid intima-media thickness. Results: The LDL particle size was found to be significantly correlated with insulin resistance using a simple Pearson’s correlation(r=0.233, P<0.01), but the independent predictors of the LDL particle size, as determined by a multiple stepwise regression analysis, were serum triglyceride(TG), high density lipoprotein(HDL) cholesterol level and age(beta=-0.403, P=< 0.001; beta=0.309, P=0.003; beta=-0.219, P=0.016, respectively). Significant relationships were found between an increasing IMT and the traditional risk factors of atherosclerosis: age, LDL cholesterol, HDL cholesterol, systolic and diastolic blood pressure(r=0.490, P<0.001; r=-0.251, P<0.01; r=0.211, P<0.05; r=0.298, P<0.01; r=0.263, P<0.01, respectively). However, no significant correlation was found between an increasing IMT and the LDL particle size (r=-0.172, P=0.075). Conclusion: The best predictors for the LDL particle size were the serum TG level, HDL cholesterol level and age. Insulin resistance was not found to be an independent predictor of the LDL particle size. Small dense LDL was not found to be a predictor of the IMT in healthy Koreans.ope

Prevalence of Diabetes Mellitus(Fasting Plasma Glucose by the ADA Criteria) and Impaired Fasting Glucose according to Anthropometric Characteristics and Dietary Habits

Background: The study is based on the National Health and Nutrition Examination Survey in Korea(1998). With these data, we want to predict the prevalence of diabetes mellitus(DM) and impaired fasting glucose(IFG), By investigating anthropometric characteristics and dietary intake habits, we also wanted to analyze any significant correlation between those factors and the prevalences of DM and IFG. Methods: The study group was comprised of 8,166 people, a representative group of Koreans, who had undergone a health check-up and food intake survey among the total 39,331 members of 12,189 families who were surveyed. Results: The final results are as the follows. 1) The peak prevalence of DM was 15.92% among women in their sixties and 18.21% among men in their fifties, and that of IFG was found to be 16.27% of women in their seventies and 14.09% of men in their sixties. 2) When analyzing the eating habits and the prevalences of DM and IFG, we found that women with more glucose intake had a lesser risk of DM, but this was of no statistical significance. 3) In men, age, total cholesterol, triglyceride(TG), and hypertension(HTN) were revealed as meaningful factors and in women, age, TG, and HTN were revealed as meaningful factors. As to the IFG, in females, age and TG were meaningful factors, and in males, age, TG, the waist/hip ratio (WHR), and body mass index (BMI) were meaningful factors. Conclusion: Although this study could not demonstrate meaningful correlation between diet habits and DM, the prevalence of IFG and the recent increase in the prevalence of DM in Koreans, owing to alterations in their diet habits, demands further organized group study for a better understanding of their relationshipope