经IFN-r基因修饰并微囊化的人羊膜上皮细胞通过抑制FAK/Src激酶复合物活性拮抗瘢痕纤维化的作用及机制研究

增生性瘢痕HS是一种严重的皮肤纤维化疾病，转抗纤维化细胞因子基因的干细胞移植为HS治疗提供了新思路。IFN-r是明确抗纤维化的细胞因子，人羊膜上皮细胞hAEC易获取、低免疫原性、无致瘤性、旁分泌强大，是干细胞治疗的理想种子细胞和优良基因载体。课题组研究发现，hAEC可抑制创面愈合后期的瘢痕形成及成纤维细胞向肌成纤维细胞的分化。因此，移植经IFN-r修饰的hAEC可能是拮抗HS纤维化的一种有效策略，而微囊化又为转基因干细胞移植提供了自我更新和免疫豁免微环境。课题组还发现，抑制FAK/Src激酶活性可显著缓解ECM沉积及胶原紊乱，因此调节FAK/Src活性可能是hAEC-IFN-r拮抗HS的关键环节。课题拟通过基因重组、干细胞移植、细胞微囊化、定点突变、过表达等技术，明确hAEC-IFN-r缓释微囊拮抗HS的作用，揭示FAK介导的上下游通路参与其中分子机制，为临床治疗HS提供新策略和理论依据。

Hypertrophic scar (HS) is a serious skin fibrotic disease. Stem cell transplantation with anti-fibrotic cytokine gene provides a novel way for HS treatment. IFN-r is a definite anti-fibrotic cytokine. Human amniotic epithelial cell (hAEC), which is easy to obtain, non-tumorigenic, with low immunogenicity and strong paracrine function, is an ideal seed cell and excellent gene carrier for stem cell therapy. Our research group found that hAEC could inhibit scar formation during the late stage of wound healing and suppress the differentiation of fibroblast into myofibroblast. Therefore, the transplantation of IFN-r gene modified hAEC might be an effective strategy against HS fibrosis. In the meantime, microencapsulation provides a microenvironment that is beneficial to the self-renewing and immune privilege for stem cell transplantation. Our research group also found that inhibiting the activity of FAK/Src kinase complex significantly alleviated ECM deposition and collagen disorganization. Thus, regulating the activity of FAK/Src might be a critical step in hAEC-IFN-r against HS fibrosis. The current project aims to clarify the antagonistic effect of hAEC-IFN-r microcapsule on HS fibrosis by using techniques such as gene recombination, stem cell transplantation, cell microencapsulation, site-directed mutagenesis and gene over-expression , and reveal FAK/Src-mediated molecular mechanism. This project is expected to provide a novel strategy and theoretical basis for clinical treatment of HS.