Copland Ian B, Jolicoeur E Marc, Gillis Marc-Antoine, Cuerquis Jessica, Eliopoulos Nicoletta, Annabi Borhane, Calderone Angelo, Tanguay Jean-Francois, Ducharme Anique, Galipeau Jacques
Sir Mortimer B. Davis Jewish General Hospital, McGill University, 3755 Cote Ste-Catherine Road, Montreal, Quebec, Canada H3T 1E2.
Cardiovasc Res. 2008 Aug 1;79(3):405-15. doi: 10.1093/cvr/cvn090. Epub 2008 Apr 8.
Mesenchymal stromal cells (MSCs) possess intrinsic features that identify them as useful for treating ischaemic syndromes. Poor in vivo survival/engraftment of MSCs, however, limits their overall effectiveness. In this work, we tested whether genetically engineering MSCs to secrete erythropoietin (Epo) could represent a better therapeutic platform than MSCs in their native form.
MSCs from C57Bl/6 mice were retrovirally transduced with either an empty vector or one that causes the production of Epo and were then analysed for the alterations in angiogenic and survival potential. Using a mouse model of myocardial infarction (MI), the regenerative potential of null MSCs and Epo-overexpressing MSCs (Epo+MSCs) was assessed using serial echocardiogram and invasive haemodynamic measurements. Infarct size, capillary density and neutrophil influx were assessed using histologic techniques. Using in vitro assays coupled with an in vivo Matrigel plug assay, we demonstrate that engineering MSCs to express Epo does not alter their immunophenotype or plasticity. However, relative to mock-modified MSCs [wild-type (WT)-MSCs], Epo+MSCs are more resilient to apoptotic stimuli and initiate a more robust host-derived angiogenic response. We also identify and characterize the autocrine loop established on MSCs by having them secrete Epo. Furthermore, in a murine model of MI, animals receiving intracardiac injections of Epo+MSCs exhibited significantly enhanced cardiac function compared with WT-MSCs and saline-injected control animals post-MI, owing to the increased myocardial capillary density and the reduced neutrophilia.
Epo overexpression enhances the cellular regenerative properties of MSCs by both autocrine and paracrine pathways.
间充质基质细胞（MSC）具有一些内在特性，使其被认为可用于治疗缺血综合征。然而，MSC在体内的低存活率/植入率限制了它们的整体疗效。在本研究中，我们测试了通过基因工程使MSC分泌促红细胞生成素（Epo）是否能比天然形式的MSC代表更好的治疗平台。
用空载体或能促使产生Epo的载体对C57Bl/6小鼠的MSC进行逆转录病毒转导，然后分析其血管生成和存活潜能的变化。使用心肌梗死（MI）小鼠模型，通过连续超声心动图和有创血流动力学测量评估空载MSC和Epo过表达MSC（Epo+MSC）的再生潜能。使用组织学技术评估梗死面积、毛细血管密度和中性粒细胞浸润情况。通过体外试验结合体内基质胶栓塞试验，我们证明使MSC表达Epo不会改变其免疫表型或可塑性。然而，相对于模拟修饰的MSC[野生型（WT）-MSC]，Epo+MSC对凋亡刺激更具抵抗力，并引发更强有力的宿主来源的血管生成反应。我们还鉴定并表征了MSC通过分泌Epo建立的自分泌环。此外，在MI小鼠模型中，与WT-MSC和MI后注射生理盐水的对照动物相比，接受心内注射Epo+MSC的动物表现出显著增强的心脏功能，这归因于心肌毛细血管密度增加和中性粒细胞减少。
Epo过表达通过自分泌和旁分泌途径增强了MSC的细胞再生特性。
