초록 ▼

당뇨병에서 유용한 바이오마커를 발굴하기 위해 지방세포, 혈관/염증세포, 간 등에서 발현되어 분비되는 단백질을 스크리닝할 수 있도록 Genome-wide secretome을 확보하고 적용함. 지방조직에서 유래한 Serpina3C를 발굴하여 기능분석을 시행함. 췌장, 지방, 경부당부하검사 혈청, 임상정보를 포함하는 인체유래물 코호트를 성공적으로 구축함. 베타세포 특이 Ang1 KO 생쥐에서 glucagon intolerance가 발생함을 규명하고, 해당 생쥐에서 분리된 췌도에서 유전체 분석 후 Reg1, Reg2를 췌도 재생에 관여하는

당뇨병에서 유용한 바이오마커를 발굴하기 위해 지방세포, 혈관/염증세포, 간 등에서 발현되어 분비되는 단백질을 스크리닝할 수 있도록 Genome-wide secretome을 확보하고 적용함. 지방조직에서 유래한 Serpina3C를 발굴하여 기능분석을 시행함. 췌장, 지방, 경부당부하검사 혈청, 임상정보를 포함하는 인체유래물 코호트를 성공적으로 구축함. 베타세포 특이 Ang1 KO 생쥐에서 glucagon intolerance가 발생함을 규명하고, 해당 생쥐에서 분리된 췌도에서 유전체 분석 후 Reg1, Reg2를 췌도 재생에 관여하는 분비단백질 후보로 도출함. 간세포, 마우스 간조직, 당뇨 환자의 간조직에서 genome-wide transcriptome, secretome 분석을 하여 영양분 수치와 인슐린 저항성에 민감하게 변화하는 신규 바이오마커 hepatokine 후보 Gdf15, Fos, Cyr61를 규명하였음. 이들 후보가 인슐린 감도, 당 항상성에 선택적으로 미치는 영향을 밝히고 바이오마커로서의 가능성을 평가하였음.


(출처 : 보고서 요약서 3P)

Abstract ▼

Purpose & Contents
< Purpose >
The final aim of this project is to develop biomarkers or disease targets of diabetes, which will be useful for the development, progress, analysis of disease control, and prognosis, using an analysis by genome-wide secretome. For this purpose, 1) we will develop

Purpose & Contents
< Purpose >
The final aim of this project is to develop biomarkers or disease targets of diabetes, which will be useful for the development, progress, analysis of disease control, and prognosis, using an analysis by genome-wide secretome. For this purpose, 1) we will develop a way of analysis using genome-wide secretome and set up the database, 2) we will obtain secretome database from liver, adipose tissue, inflammatory cells, endothelial cells, and pancreas in cells-animals-humans, 3) we will investigate wether the candidate genes can be used for biomarkers or disease targets.
< Contents >
< First year >
1) A development of genome-wide secretome : By using algorithm in human and mouse gene database, we will construct genome-wide secretome.
2) A construction of human diabetes cohort : Human tissue bank will be used for the construction of liver, adipose tissues, pancreas, endothelial and inflammatory cells to obtain tissues and clinical information.
3) An identification of new biomarkers : We will identify new biomarkers from various tissues, which will be investigated in next year.
< Second year >
1) An identification of new biomarkers using genome-wide secretome : Genome-wide secretome will be used for the identification of new biomarkers and we will confirm whether they are produced and secreted from tissues.
2) An analysis and expension of human diabetes cohort : we will also obtain gene expression database from the samples of diabetes cohort and analyze them for new biomarkers.
3) An investigation of biological importance and relevance of biomarkers : We will analyze the candidate proteins whether they can be used for diabetic biomarkers, by investigating their biological roles.
< Third year >
A usefulness of biomarkers for diabetes : Using human diabetes cohort, we will adopt an analysis of genome-wide secretome, and finally will integrate the biomarkers to develop the "new biomakers of diabetes", which will be used for the development, progress, analysis of disease control, and prognosis.

Results
1. Development of human-mouse genome-wide secretome database.
1) We developed a secretome database, 396 human protein genes are defined as "human genome-wide seretome".
2) In adipocyte model using conditioned medium, a set of transcriptome database was obtained. It is integrated with the genome-wide secretome, releasing many candidate genes for new biomarkers. Of these, we investigated Serpina3C as a strong candidate biomakers, for its biological role and relationship to diabetes.
2. Developing human-derived-tissue cohort for diabetes study
1) In Gangnam Severance Hospital, we constructed 120 subjects of pancreatectomy patients cohort composed with pancreas tissue, omental/subcutaneous fat tissue, serum after OGTT, and all the clinical information.
2) We also constructed 200 subjects of diabetes serum corhort with full diabetic complication study data
3. Developing novel biomarkers on diabetes targeting Inflammatory/vascular part.
1) We found that beta cell specific Angiopoietin (Ang1) KO mice develop glucose intolerance because of the decreased insulin secretion from islets. In the transcriptome and secretome analysis from microarray data on islets from Ang1 KO mice, we could find Reg1 and Reg2 as the candidate molecules for islet regeneration/proliferation in diabetic setting in mice
2) In the pancreatectomy corhot, we demonstrated that the development of diabetes in pancreatic cancer patients is mainly originated from impaired insulin secretion by some factors secreted from tumor tissue
3) We demonstrated that relative glucagon-to-insulin ratio can be a parameter for uncontrolled diabetes in nonobese long duration diabetic patient.
4. Identification of hepatokine candidate genes, which their expression is sensitive to nutrient levels.
1) In response to starvation, expressions of Pgcd, Pgat1, Fgf4, Egfb4, selectoprotein, Angiopoietin-like 4 were elevated in primary mouse hepatocytes.
2) In contrast, upon nutrient rich condition, expressions of lcit1, Bdnf, Bmp2 were increased in mouse hepatocytes.
3) After treatment of metformin which reduces insulin resistance, expression of Gdf15, Fst, Fos, Hmgb2, Cyr61, Lgals3, Endod1, Cldn1, Id3, Ctgf were elevated in mouse hepatocytes, suggesting those gene expression are associated with insulin sensitivity.
5. Genome-wide secretome analysis to identify novel hepatokine candidate genes related to insulin resistance.
1) The expression of Gdf15 gene was elevated in response to insulin and metformin. When Gdf15 is depleted, the expression of gluconeogenic gene was increased, suggesting that, hepatic Gdf15 regulates gluconeogenesis.
2) Expression of Hmgb2, Endod1, Cldn1, Cyr61, Id3, Ctgf was decreased in livers from high fat diet-fed mice.
5. Analysis of the potential relevance of novel hepatokine as a biomarker and therapeutic target for the development of diabetes : The expression of GDF15, Fos, Cyr61 were elevated in liver tissues from diabetic subjects compared to non-diabetic subjects.

Expected Contribution
1) New biomarkers of diabetes will pave a way for the development, progress, analysis of disease control, and prognosis.
2) This project will contribute in identifying and developing a new therapeutic targets and drugs, as well as new theories of diabetes.
3) The cohort which we developed as diabetic patient-derived-tissue bank is unique and valuable cohort that all fat/pancreas/serum were collected from each single subject. The cohort will be used to find new targets in diabetes which is applicable for human.
4) The candidate molecule like Reg1 and Reg2 which was developed in Ang KO mice study, and new parameter such as glucagon-to-insulin ratio can be a new target for diabetes treatment and further study will be necessary.
5) The knowhow to develop human cohort, sorting out intra-islet endothelial cell, and doing RNAseq with small amount of RNA which were build up during this study will be applied for next step study.
6) This study will provide a physiological basis to identify molecular targets with great potential value in the development of biomarker and therapeutic strategies to treat insulin resistance in diabetes.


(출처 : SUMMARY 7P)

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 국문 요약문 ... 4
• SUMMARY ... 7
• 목차 ... 9
• 1. 연구개발과제의 개요 ... 10
• 1-1. 연구개발 목적 ... 10
• 1-2. 연구개발의 필요성 ... 11
• 1-3. 연구개발 범위 ... 13
• 2. 국내외 기술개발 현황 ... 14
• 3. 연구수행 내용 및 결과 ... 18
• 4. 목표달성도 및 관련분야 기여도 ... 55
• 4-1. 목표달성도 ... 55
• 4-2. 관련분야 기여도 ... 57
• 5. 연구결과의 활용계획 ... 58
• 6. 연구과정에서 수집한 해외과학기술정보 ... 59
• 7. 연구개발성과의 보안등급 ... 59
• 8. 국가과학기술종합정보시스템에 등록한 연구시설·장비 현황 ... 59
• 9. 연구개발과제 수행에 따른 연구실 등의 안전조치 이행실적 ... 59
• 10. 연구개발과제의 대표적 연구실적 ... 60
• 11. 기타사항 ... 60
• 12. 참고문헌 ... 61
• 별첨4. 실적 증빙자료 ... 62
• 별첨5. 기타 첨부서류 ... 74
• 끝페이지 ... 84