임상가용한 X-연관 부신백질이 영양증역분화 줄기세포 유래 신경전구 세포의 자격화 및 생체재료를 이용한 효율적 세포치료법 개발 = Development of optimizing cell therapy by using clinically applicable and qualified neural precursors derived from X-linked adrenoleukodystrophy reprogramming stem cells and functional biomaterial scaffolds

다국어 초록 (Multilingual Abstract)

Purpose&Contents
Purpose : At present, no satisfactory therapy for X-ALD is available. Recently, the induced pluripotent stem cell (iPSC) technique is being developed and has used for human disease. In this study, we established disease- and patient-specific reprogrammed cells from this disease that are rare and known to be incurable, subsequently differentiate into oligodendrocyte precusor cells. Our aims are to generate the non-viral gene transfer method, and ABCD1 correction system for cell therapy using TALEN or Crispr-Cas9. Functional confirmation of iPS-derived oligodendrocyte precursor cells by cell-transplantation into animal models in order to identify their therapeutic potential. Establishment of patients derived-iPSC technology will lead us in providing platforms for addressing mechanisms of diseases and in development of new drugs.
Contents
- Establishment of iPS cells from X-ALD patients by using 4reprogramming factors.
- Development of hydrogel based bio-materials that mimic human cell matrix - Development of purification method of PSA-NCAM+ or A2B5+ cells and advancement of oligodendrocyte differentiation protocol from PSA-NCAM+ or A2B5+ cells.
- Generation unharmful iPS cells by utilizing non-viral and non-integrating techniques after correction of defect ABCD1 by TALEN or Crispr-Cas9.
- Functional confirmation of iPS-derived oligodendrocyte precursor cells by cell-transplantation into animal models with hydrogel based bio-materials.
Results
- Establishment of iPSC lines from ABCD1 gene mutated X-ALD patient
- Confirm of pluripotency in acquired iPSC lines
- Differentiation of oligodendrocyte precursor cells
- Developemnet of ABCD1 correction systems for cell therapy using TALEN and Crispr-Cas9
- Development of hydrogel based bio-materials that mimic human cell matrix: 1) hydrogel which Brain extracellular matrix based 2) hydrogel which hyaluronic acid based
- Analysis for biocompatibility and cell-promotional function of bio-materials
- Functional confirmation of iPS-derived oligodendrocyte precursor cells by cell-transplantation into animal models with hydrogel based bio-materials.
Expected Contribution : Development of disease-specific stem cells for clinics and hydrogel base bio-materials can be applied to other diseases. Therfore these bio-technologies could offer a good clinic strategy and economic value added.

국문 초록 (Abstract)

연구의 목적 및 내용
1. 임상가용한 X-ALD 역분화줄기세포주 수립
2. 생체모사 기능성 3D 하이드로젤 제작 및 특성 분석
3. 신경전구세포로의 분화와 순수분리 및 증식 배양
4. 하이드로젤 기반 신경전구세포 이식 후 세포 생착 및 분화능 조사
5. 동물모델에서 하이드로젤 기반 세포치료에 의한 치료 효능 및 안전성 검증
연구개발성과
1. episomal vector를 이용한 임상가용한 X-ALD 역분화줄기 세포주 확립
2. 역분화줄기 세포를 이용하여 환자유래 희돌기아교세포 분화확립
3. X-ALD 유전자교정용 TALEN과 Crispr/Cas-9 유전자 가위 2종 개발
4. 생체모사 기능성 3D 하이드로젤 2종제작: Brain extracellular matrix 하이드로젤, Hyaluronic acid 하이드로젤
5. 하이드로젤의 생체적합성 분석
6. 하이드로젤의 신경세포 분화촉진 기능성 확인
7. 신경전구세포의 희소돌기교세포로의 분화 촉진 및 myelination 촉진 표면 개발
8. 동물모델에서 하이드로젤 기반 신경전구세포 이식 후 세포 생착 및 분화능 조사
연구개발성과의 활용계획(기대효과) : 임상가용한 질병 특이 줄기세포주 확립과 이식세포의 자격화 및 개발된 하이드로젤 생체재료는 다른 종류의 세포 이식술에도 적용할 수 있으므로, 임상적용 응용범위를 확대하여 세포이식용 세포와 생체재료의 상업화 및 기술이전으로 경제적 이익과 부가가치 창출이 가능함.