초록 ▼

본 연구는 줄기세포 프라이밍 기술을 활용한 폐동맥고혈압 줄기세포 치료 효능 증진을 최종 목표로 하고 있음. 조혈모줄기세포 프라이밍 선행 연구에서 검증한 sphingosine-1 phosphate (S1P)과 ceramide-1 phosphate (C1P)와 같은 bioactive lipid와 LL-37 cationic peptide 물질 전처리를 통한 다양한 기원의 사람 중간배엽줄기세포 (AD-MSC, UCB-MSC) 특성 변화를 연구하고, monocrotaline 주사 및 저산소 유도 폐동맥고혈압 동물 모델을 통하여 줄기세포 프라

본 연구는 줄기세포 프라이밍 기술을 활용한 폐동맥고혈압 줄기세포 치료 효능 증진을 최종 목표로 하고 있음. 조혈모줄기세포 프라이밍 선행 연구에서 검증한 sphingosine-1 phosphate (S1P)과 ceramide-1 phosphate (C1P)와 같은 bioactive lipid와 LL-37 cationic peptide 물질 전처리를 통한 다양한 기원의 사람 중간배엽줄기세포 (AD-MSC, UCB-MSC) 특성 변화를 연구하고, monocrotaline 주사 및 저산소 유도 폐동맥고혈압 동물 모델을 통하여 줄기세포 프라이밍 전임상 효능 평가를 실시하였음. 세포 배양 모델을 통하여, S1P, LL-37, C1P, 5-Aza 저분자 물질과 Nurr1 유전자 과발현 기반 UCB-MSC priming은 줄기세포의 이동성, 자가재생능력, 항염증반응을 유의적으로 증진시키며, S1P primed MSC는 단순배양한 UCB-MSC보다 폐동맥고혈압 치료 효능이 증진됨을 폐동맥고혈압 동물 모델에서 확인하였음. 또한 비침습적 줄기세포 치료 효능 모니터링과 기전 연구를 위한 optical imaging system, 동물 microPET, 단일세포 발현체 분석법을 확립하였음. 따라서 본 연구를 통하여, MSC priming을 통한 치료 효능 증진이라는 개념증명 (proof of concept, PoC)와 이를 활용한 폐동맥고혈압 치료 기술에 대한 임상적 진입 근거 확보하였음.

Abstract ▼

Over the past decade, the treatment of PAH has evolved considerably as a deeper understanding of underlying pathogenesis. However, mortality remains high despite of these treatments. Stem cell (SC) therapy has become a potential therapy for pulmonary artery hypertension (PAH). However, simple cultiv

Over the past decade, the treatment of PAH has evolved considerably as a deeper understanding of underlying pathogenesis. However, mortality remains high despite of these treatments. Stem cell (SC) therapy has become a potential therapy for pulmonary artery hypertension (PAH). However, simple cultivated stem cells exhibit several technical hurdles for their application into clinic, especially their poor engraftment capacity. In this study, we investigate the role of a bioactive lipid, S1P and a cationic peptide, LL-37 on the priming of MSCs which have been popularly employed in stem cell therapeutics and finally prove the beneficial function of MSC priming in vivo condition using experimental PAH model.
In ex vivo-expanded MSCs with S1P, MSC showed stronger chemoattraction and phosphorylation of both MAPKp42/44 and AKT (involved in signaling transduction for SC migration). S1P priming also potentiated its self renewal activity, presented by increased colony forming unit in fibroblast assay and anti-inflammation capacity.
Examining effect of MSCs priming in vivo condition using a PAH animal model induced by MCT. PAH was induced in rats by monocrotaline (MCT) injection. Human adipose or cord blood derived MSC were injected via tail vein at 2weeks after MCT injection. MCT injection elevated right ventricular systolic pressure (RVSP, 42.0± 7.4mmHg vs. 21.6± 2.6mmHg, P<0.001). Human CB-MSC or S1P primed CB-MSC attenuated this RVSP elevation significantly (32.4± 4.0mmHg and 30.1± 4.4 mmHg, respectively; P<0.05). S1P primed CB-MSC, not CB-MSC without priming, also showed significant reduction in right ventricular weight ratio and pulmonary vascular wall thickness. Meanwhile, ADMSC failed to show improvement in these variables. When we examined the gene expression of genes related to angiogenesis and anti-inflammation in lung tissues, S1-MSC group expressed the highest level of transcripts for rat SDF-1 and its unique receptor CXCR4. In addition, the expression levels of HGF, cMet receptor, and PDGFFs; PDGFa and -b and their receptors were increased in the S1P-MSC group.
Taken together, UCB-MSC had some possibilities in treatment of PAH, and S1P priming increased the effects of SC therapy by improvement in cardiac and vascular remodeling. By titration of optimal protocol in further study, stem cell therapy can have the basis of clinical trials in human PAH treatment. Furthermore, our finding could provide a novel strategy to improve therapeutic efficacy of the adult stem cell based therapy for several complex incurable disorders.