초록 ▼

생체 모방 환경에서의 공배양 시스템을 활용하여 조골 및 파골세포의 조절에 관여하는 신규 분자를 찾아내고 그 작용 기전을 in vitro 및 in vivo에서 규명함으로써 골격계 질환의 치료제 개발에 유용한 분자 타겟을 발굴함을 연구 목표로 하였음. 칼슘포스페이트 배양기에서 조골-파골세포의 공배양을 실시하고 DNA 마이크로어레이로 575개 유전자의 발현 변화를 찾아내었음. 변화한 유전자 중 ApoE, MSK1, PMCA, Nrf2 등을 선별하고 조골, 파골세포에서 유전자 knock-down과 과발현 구현 후 세포 분화, 활성분석과 신

생체 모방 환경에서의 공배양 시스템을 활용하여 조골 및 파골세포의 조절에 관여하는 신규 분자를 찾아내고 그 작용 기전을 in vitro 및 in vivo에서 규명함으로써 골격계 질환의 치료제 개발에 유용한 분자 타겟을 발굴함을 연구 목표로 하였음. 칼슘포스페이트 배양기에서 조골-파골세포의 공배양을 실시하고 DNA 마이크로어레이로 575개 유전자의 발현 변화를 찾아내었음. 변화한 유전자 중 ApoE, MSK1, PMCA, Nrf2 등을 선별하고 조골, 파골세포에서 유전자 knock-down과 과발현 구현 후 세포 분화, 활성분석과 신호전달경로 분석을 통해 선별된 유전자의 뼈세포 조절 작용기전을 in vitro상에서 규명하였음. 또한 낙아웃 마우스와 골소실 모델 동물의 뼈에 대한 micro-CT 분석 및 조직형태계측학적 평가를 실시하여 in vivo 기능을 검증하였음. 연구결과 IF 10이상 1편을 포함, 4편의 SCI 논문으로 발표하였으며 1건의 특허를 등록하였고 2명의 박사인력을 양성 하였음.

Abstract ▼

I. Purpose and Needs
- Medical expenses for bone diseases greatly increase as the population gets old.
- Development of reagents with new mechanisms of action is needed as current therapeutics have limits in their effectiveness and complications.
- This study is to discover new regulators o

I. Purpose and Needs
- Medical expenses for bone diseases greatly increase as the population gets old.
- Development of reagents with new mechanisms of action is needed as current therapeutics have limits in their effectiveness and complications.
- This study is to discover new regulators of osteoblasts and osteoclasts by using a bio-mimicking coculture system, to find the mechanism of action in vitro and to verify their role in vivo using animal models.
Ⅱ. Specific Aims
- Establishment of protocols for osteoblast and osteoclast coculture
- Establishment of culture system with calcium phosphate-coated dises
- Discovery of new molecules by genomics approaches
- Selection of new candidate molecules by bioinformatics analyses
- Analyses of expression patterns of new molecules during OB and OC differentiation
- Evaluation of effects of gene knock-down on OB and OC
- Analyses of signaling pathways of new molecules
- Investigation on mechanisms for the regulation of expression of new molecules
- Analyses of effects of new molecule modulators on in vivo bone resorption and OC formation
- Analyses of effects of modulators on in vivo bone formation and OB differentiation
Ⅲ. Results
- OB-OC coculture system on calcium phosphate dises was established and 575 genes were found to diffentially expressed via DNA microarray analyses.
- Among the differentially expressed genes, some selected genes including ApoE, MSK1, PMCA, and Nrf2 were further analyzed for their expression during OB and OC differentiation.
- In vitro mechanisms of action was investigated by examining the cell differentaion and activation and signal transduction pathways after gene knock-down or overexpression.
- The in vivo roles of selected genes were evaluated by micero-CT and histomorphometric analyses of bones of knock-out and bone erosion mice models.
- 4 SCI papers including 1 with more than 10 of impact factor were published and 1 patent was registered. 2 PhD were produced.
Ⅳ. Future Directions
- Validation of target effectiveness through further in-depth studies
- Evaluation of application to RA and bone metastases models