초록 ▼

목적 및 방법: 미토콘드리아 기능이상과 근감소증이 노인성 당뇨병 발생의 대사 지표와 관련성을 보이는지 확인함. 노화표적 단백질을 발굴하여 기능을 평가하고, 미토콘드리아 기능 이상을 개선하는 신약 타겟의 효과를 검증함.

결과: 노인에서 근감소증과 복부 비만이 동시에 있을 경우 인슐린저항성이 증가하고 당뇨병을 포함한 심혈관 대사질환의 위험이 증가함. 노인 코호트 추적 관찰을 통해 혈당 조절이 불량한 경우 근육량의 변화없이 근력의 감소가 가속화됨을 확인함. 더불어 근육량이 적은 당뇨병 노인의 혈액과 근육조직에서 표지자로서 C

목적 및 방법: 미토콘드리아 기능이상과 근감소증이 노인성 당뇨병 발생의 대사 지표와 관련성을 보이는지 확인함. 노화표적 단백질을 발굴하여 기능을 평가하고, 미토콘드리아 기능 이상을 개선하는 신약 타겟의 효과를 검증함.

결과: 노인에서 근감소증과 복부 비만이 동시에 있을 경우 인슐린저항성이 증가하고 당뇨병을 포함한 심혈관 대사질환의 위험이 증가함. 노인 코호트 추적 관찰을 통해 혈당 조절이 불량한 경우 근육량의 변화없이 근력의 감소가 가속화됨을 확인함. 더불어 근육량이 적은 당뇨병 노인의 혈액과 근육조직에서 표지자로서 C1QTNF5, SFRP5, PRDM16, FGF21, PLA2G7을 발굴함. 특히 C1QTNF5의 경우 유산소 운동을 통해 혈중 농도가 감소하고, C1QTNF5의 농도와 mtDNA density가 음의 상관성을 확인함. 당뇨병 환자 골격근의 단백질발현 통합분석을 통해 미토콘드리아 기능 관련 표적 표지후보단백질들을 발굴하였으며 미토콘드리아 기능이상과 당뇨병의 새로운 연결고리로써 RXRalpha의 역할을 규명하였고, 당뇨병에서 ERK-cMyc 경로를 통한 미토콘드리아 단백발현조절의 이상을 규명함. 미토콘드리아 기능개선 후보물질로서 천연물 스크리닝을 통해 발굴한 2’4’- dihydroxy- 6’-methoxy -3’5’- dimethylchalcone(DMC)와 ginsenoside Rg3, L-carnitine, Coenzyme Q10의 기능과 연관 기전을 확인하고, 노화 당뇨병 동물 모델에서 DMC, L-carnitine과 Rg3의 인슐린 저항성 및 미토콘드리아 기능 개선 효과를 확인함. 또 미토콘드리아 기능을 회복하는 표적으로서 SENP2를 확인하고 세포 및 동물모델에서 검증함.

( 출처 : 요약서 3p )

Abstract ▼

Ⅳ. Research and development results
1. Aging cohort study about sarcopenia and its link with metabolic deterioration
• In KLoSHA study, 287 men and 278 women aged 65 or older were recruited. They were evaluated with standardized clinical evaluation and blood samples were collected.
• The HO

Ⅳ. Research and development results
1. Aging cohort study about sarcopenia and its link with metabolic deterioration
• In KLoSHA study, 287 men and 278 women aged 65 or older were recruited. They were evaluated with standardized clinical evaluation and blood samples were collected.
• The HOMA-IR of subjects with sarcopenic obesity was higher and they were at higher risk for metabolic syndrome (odds ratio 8.28 [95% CI 4.45- 15.40]). We established parameters and cut-off values of sarcopenia and abdominal obesity.
• Plasma adiponectin levels and severity of sarcopenic obesity showed negative correlation,and lower adiponectin with elevated RBP4 levels increased cardiometabolic risk as much as 6 times higher.
• On 2nd wave of the KLoSHA, subjects with HbA1c over 8.5% showed decrease in muscle mass. Sarcopenia defined by muscle mass and handgrip strength predicted 3.4-fold increase in mortality, and sarcopenia defined by muscle mass, handgrip strength,and walking velocity predicted 5.6-fold increase.

2. Investigation of mitochondrial dysfunction and discovery of biomarkers in elderly diabetes
• Decrease in quantity and quality of mitochondrial function was observed in patients with diabetes. This change was more significant in elderly patients; additionally their mitochondria were morphologically destroyed.
• In elderly diabetes, blood levels of IL-6 was increased. OLETF, insulin resistant animal model, showed increased C1QTNF5 concentration in DNA-injured muscle tissue.C1QTNF5 might be an indicator mediating mitochondrial dysfunction and insulin resistance. Clusterin, nampt(visfatin), GPx3 levels were comparable but PLGA2G7 was higher in blood samples of elderly diabetes compared to elderly non-diabetics. In addition, FGF21, PRDM16, SFRP5, and C1QTNF5 were decreased. In addition, C1QTNF5, SFRP5, PRDM16, and FGF21 levels were increased and Follistatin-like 1 level was decreased in muscle samples of elderly diabetes compared to elderly non-diabetic counterpart. C1QTNF5, SFRP5, PRDM16, and FGF21 levels were increased and Follistatin-like 1 level was decreased in elderly diabetes compared to elderly non-diabetic counterpart. Therefore, these markers might have a role as biomarkers for aging diabetes.
• After aerobic exercise, C1QTNF5 significantly decreased in young and elderly women. mtDNA was initially lower in elderly women than young women, but it increased significantly after exercise. In aging model of Fisher Rat, reversal of increased muscular cytokine, decreased catalase/Gpx, increased ROS were demonstrated by exercise. Also, expression of gene coding mitochodrial ribosome decreases in elderly, but increases with exercise. In aging animal model, myostatin was increased and follistatin was decreased after exercise.

3. Discovery of targets about mitochondrial dysfunction through muscle tissue analysis and invention of novel drug targets
• By analyzing the gene expression, using affymetrix chip, in the skeletal muscle of human and animal models, candidate genes were found as follows. Elderly group vs.young group (Human- 473 count increase, 180 count decrease in elderly gene expression; SD rat- 2224 count increase, 2565 count decrease in elderly gene expression) Elderly control group vs. diabetes group (Diabetes group- 2336 count increase, 2600 count decrease). The candidate gene related to the muscle mass or mitochondria has shown an increase in clusterin, PLA2G7, MYH3, MYH10, and GPx3 while showing a decrease in myostatin, IRS, UCP3, ARRDc2 and no significant change in Sirts in the elderly group.
• LC-MS/MS proteome profiling was adopted to evaluate muscle tissues between diabetes and normal counterpart. Total 2,179 proteins were isolated and we tested their expression in muscle tissue, related genetic changes, and cellular mechanisms. Mitochondrial MRM (multiple reaction monitoring) system revealed over 300 proteins related with mitochondria and they were validated with patients' samples.
• Microarray meta-analysis using 5 human muscle tissues showed hub regulator gene sets which could regulate insulin resistance in muscle tissue. In elderly diabetes, proteins related with mitrochondria were decreased which was similar to genetic expression of mitochondria. The unbiased integrative approach of transcriptional profiles revealed ERK-c-Myc pathways as a novel pathway for skeletal muscle insulin resistance. Also,RXRα overexpression or activation alleviates insulin resistance by increasing IRS1 expression.

4. Development of therapeutics improving mitochondrial function and evaluation the effects on animal models
• Flag-HA-nampt expression sequence was inserted into adenoviral shuttle vector (pAd Track CMV) and expression of nampt was validated. Using this system 50 products from ginseng leaf(G1-13), plant compounds(C1-7), ginsenoside (cpp501-530) and over 500 products from natural extracts were tested. Finally C5 from Cleistocalyx operculatus and Rg3 form ginseng were discovered for novel drug targets.
• Vietnam plant-derived, C5, Cleistocalyx operculatus, increased PPARɣ activity. Increase in glucose uptake in muscle and improvements in insulin resistance by accelerating fatty acid oxidation was observed.
• Injection of Rg3 increased mitochondrial extent, and increase in associated genes such as PGC1α, NRF-1, and Tfam were observed. Rg3 improves both quantity and quality of mitochondria, thereby improving insulin resistance.
• Injection of L-carnitine improved glycemic profile, and increase in autophagy/mitophagy resulted in increased mitochondrian ATP synthesis and improvement in fatty acid oxidation. The effect of L-carnitine was also evaluated in an aging diabetes animal model.
• Coenzyme Q10 administration showed decreased glucose excursion and its effect on mitochondrial function and insulin resistance were evaluated.
• In overexpression of SENP2 (SENP2 transgenic mouse, TG), increased expression of genes for fatty acid oxidation, decreased weight and fat mass, improvements in glycemic indices and insulin resistance were observed. Mitochondrial extent was significantly increased and intramyocellular fat was also decreased.

( 출처 : SUMMARY 10p )

목차 Contents

• 표지 ... 1제 출 문 ... 2보고서 요약서 ... 3요 약 문 ... 4SUMMARY ... 9C O N T E N T S ... 14목차 ... 15제1장 연구개발과제의 개요 ... 16 제1절. 연구개발의 목적 ... 16 제2절. 연구 개발의 필요성 ... 17 제3절. 연구 개발의 범위 ... 21제2장 국내외 기술개발 현황 ... 25제3장 연구개발수행 내용 및 결과 ... 27 제1절. 전단계 연구개발수행 내용 및 결과 ... 27 제2절. 미토콘드리아 기능 이상 회복을 통한 당뇨병 예방기술 개발 ... 35 제3절. 미토콘드리아 기능 이상의 조기진단 및 치료효과 평가 기술 개발 - 노인 당뇨병과 관련된 미토콘드리아 표지자 선별 및 검증 ... 46 제4절. 미토콘드리아 기능 조절 물질 및 기능이상을 개선하는 신약의 효과를 동물 모델에서 검증 ... 63제4장 목표달성도 및 관련분야에의 기여도 ... 81 제1절. 3단계 목표 달성도 및 관련분야 기여도 ... 81제5장 연구개발결과의 활용계획 ... 84제6장 연구개발과정에서 수집한 해외과학기술정보 ... 84제7장 연구시설·장비 현황 ... 84제8장 참고문헌 ... 84끝페이지 ... 87