Laboratory of Biology, School of Medicine, National and Kapodistrian University of Athens, Athens, Greece.
Biotechnology Laboratory, Centre of Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.
EBioMedicine. 2019 Jul;45:542-552. doi: 10.1016/j.ebiom.2019.07.009. Epub 2019 Jul 12.
Human mesenchymal stem/stromal cells (MSCs) and their secreted molecules exert beneficial effects in injured tissues by promoting tissue regeneration and angiogenesis and by inhibiting inflammation and fibrosis. We have previously demonstrated that the therapeutic activity of fetal MSCs derived from amniotic fluid (AF-MSCs) and their hepatic progenitor-like cells (HPL) is mediated by paracrine effects in a mouse model of acute hepatic failure (AHF).
Herein, we have combined proteomic profiling of the AF-MSCs and HPL cell secretome with ex vivo and in vivo functional studies to identify specific soluble factors, which underpin tissue regeneration in AHF.
The anti-inflammatory molecule Annexin-A1 (ANXA1) was detected at high levels in both AF-MSC and HPL cell secretome. Further functional analyses revealed that the shRNA-mediated knock-down of ANXA1 in MSCs (shANXA1-MSCs) decreased their proliferative, clonogenic and migratory potential, as well as their ability to differentiate into HPL cells. Liver progenitors (oval cells) from AHF mice displayed reduced proliferation when cultured ex vivo in the presence of conditioned media from shANXA1-MSCs compared to control MSCs secretome. Intra-hepatic delivery of conditioned media from control MSCs but not shANXA1-MSCs reduced liver damage and circulating levels of pro-inflammatory cytokines in AHF.
Collectively, our study uncovers secreted Annexin-A1 as a novel effector of MSCs in liver regeneration and further underscores the potential of cell-free therapeutic strategies for liver diseases. FUND: Fondation Santé, GILEAD Asklipeios Grant, Fellowships of Excellence - Siemens, IKY, Reinforcement of Postdoctoral Researchers, IKY.
人骨髓间充质干细胞(MSCs)及其分泌的分子通过促进组织再生和血管生成以及抑制炎症和纤维化,对受损组织发挥有益作用。我们之前已经证明,源自羊水(AF-MSCs)和其肝祖细胞样细胞(HPL)的胎儿 MSCs 的治疗活性是通过旁分泌作用在急性肝衰竭(AHF)的小鼠模型中介导的。
在此,我们结合了 AF-MSCs 和 HPL 细胞分泌体的蛋白质组学分析,以及体外和体内功能研究,以鉴定特定的可溶性因子,这些因子是 AHF 中组织再生的基础。
在 AF-MSC 和 HPL 细胞分泌体中均检测到高水平的抗炎分子膜联蛋白 A1(ANXA1)。进一步的功能分析表明,MSC 中的 shRNA 介导的 ANXA1 敲低(shANXA1-MSCs)降低了它们的增殖、克隆形成和迁移能力,以及分化为 HPL 细胞的能力。与对照 MSC 分泌体相比,来自 AHF 小鼠的肝祖细胞(卵圆细胞)在体外培养时,当存在来自 shANXA1-MSCs 的条件培养基时,其增殖能力降低。与对照 MSC 条件培养基相比,内源性肝内递送条件培养基可降低 AHF 中的肝损伤和循环促炎细胞因子水平。
总之,我们的研究揭示了分泌型 Annexin-A1 是 MSC 促进肝再生的一种新型效应因子,并进一步强调了细胞游离治疗策略在肝脏疾病中的潜力。
Santé 基金会、GILEAD Asklipeios 赠款、西门子卓越研究员奖学金、IKY、博士后研究员加强、IKY。