CRISPR/Cas9结合软骨发育不全 (ACH) 病人脂肪MSCs及其iPSCs对ACH治疗的研究

There still is no effective treatment for ACH. However, molecular biology, stem cells and regenerative medicine offer a new way for the clinical treatment of it. Up to now, there is no research report of gene-editing technique, MSCs and cell therapy in ACH. Patient adipose-dervied MSCs is an attractive source of easily-accessible adult candidate cells, which possess capscity of chondrogenic differentiation, immunoregulation, damage repair, and have not tumorigenicity. But they easily indicate senescence and don’t be cultured in long-term culture. However, adipose-derived MSCs could been more efficiently reprogrammed into iPSCs in feeder-free conditions, which avoid animal-based pollution. Reprogramming adipose-devied MSCs into iPSCs which further differentiate into MSCs would resolve the problems of limitation of transplanted cell number, and reduce the risk of iPSC tumorigenicity. Because ACH is caused by mutated FGFR3 gene, fundamental method to reapare the abnormality is to correct the mutation. It is a novel and efficient treatment for ACH to use CRISPR/Cas9-mediated FGFR3 gene correction of ACH patient adipose-derived MSCs and iPSCs and cell therapy in ACH animal model. Therefore, this project has great prospects for clinical application.

ACH为一种最常见的遗传性侏儒症，目前尚无有效治疗措施。分子与干细胞生物学及再生医学为其治疗提供新策略。至今，基因编辑技术、MSCs及细胞治疗在ACH的研究均未见报道。脂肪MSCs易获取，具有软骨分化、免疫调节、损伤修复等特性，但不能长期培养。然而，脂肪MSCs能被更高效地重编程为iPSCs、且无需小鼠饲养层维持的特性避免了动物源性污染。若将脂肪来源iPSCs分化为MSCs体内移植，不但可解决细胞数量有限的难题，且可降低iPSCs致瘤性的风险。而申请者前期在干细胞方面已取得的一些突破性研究成果为本项目奠定了基础。ACH为FGFR3基因突变所致，纠正突变基因为其治疗根本。利用CRISPR/Cas9在ACH脂肪MSCs及其iPSCs进行基因纠正，再将分化的MSCs移植于FGFR3基因突变的ACH模型小鼠进行安全性和疗效评估，这将是一种最新颖有效地治疗ACH的方法。故本项目有较大的临床应用前景。