Genetically Altered Mesenchymal Stem Cell; Paracrine Effect on Neovascularization

基因改变的间充质干细胞；

• 批准号: 7466750
• 负责人: Victor J Dzau
• 金额: $ 39万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7466750
• 关键词: Adult; Angiogenic Factor; Animals; Attenuated; Back; Biological Assay; Blood Vessels; Blood capillaries; Blood flow; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; Bone Marrow Transplantation; Cadherins; Caliber; Cardiac; Cells; Collecting Cell; Condition; Conditioned Culture Media; Coronary Occlusions; Coronary Vessels; Corrosion Casting; Data; Development; Disease; Endothelial Cells; Fibroblast Growth Factor; Ganciclovir; Gel; Gene Delivery; Genes; Grant; Green Fluorescent Proteins; Heart; Heart failure; Home environment; Hypoxia; Image; Immunologics; In Vitro; Infarction; Injection of therapeutic agent; Injury; Laboratories; LacZ Genes; Lectin; Marrow; Measures; Mediating; Mediator of activation protein; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; Messenger RNA; Modeling; Molecular; Molecular Profiling; Mus; Myocardial; Myocardial Infarction; Myocardial Ischemia; Myocardium; Pathway interactions; Phosphatidylinositide 3-Kinase Inhibitor; Play; Process; Production; Proteins; Public Health; Relative (related person); Reporter; Rodent; Role; Signal Transduction; Simplexvirus; Small Interfering RNA; Somatomedins; Staining method; Stains; Stem cells; Subfamily lentivirinae; Testing; Therapeutic; Thymidine Kinase; Transplantation; Tube; Tumor-Associated Vasculature; Vascular Endothelial Growth Factors; Week; Wild Type Mouse; angiogenesis; behavior influence; capillary; cell motility; cellular transduction; cytokine; density; design; hypoxia inducible factor 1; improved; in vitro Assay; in vivo; inhibitor/antagonist; injured; killings; neovascularization; paracrine; promoter; protective effect; repaired; suicide gene; transcription factor; vasculogenesis

DESCRIPTION (provided by applicant): We have previously demonstrated that mesenchymal stem cells (MSC), when injected into rodent hearts following myocardial infarction (MI), enhance myocardial repair and restore cardiac function. Moreover, this protective effect is enhanced by transduction of the MSC with the cytoprotective gene Akt (Akt-MSC). Furthermore, our data showed that these cells express paracrine mediators that reduce myocardial injury. In this application we hypothesize that Akt-MSC enhance myocardial repair, in part, through its paracrine effects on angiogenesis. Indeed we have shown in our laboratory that Akt-MSCs express multiple angiogenic cytokines such as VEGF and FGF. Moreover, media collected from these cells induce endothelial cell migration and tube formation in vitro. Importantly, we have recently demonstrated that Akt-MSC induces neovascularization in the infarcted heart. Since it has been shown that angiogenic signals mobilize bone marrow derived endothelial progenitor cells (EPC) that home to the ischemic myocardium, we hypothesize that (i) Akt-MSC increase angiogenesis in the post infarct heart through paracrine mechanisms and contribute to improved myocardial repair and function (ii) Akt-MSC increase angiogenesis in part through the recruitment of bone marrow derived EPCs and (iii) Akt-MSC mediated EPC recruitment is controlled by HIF 11. To test these hypotheses, we will investigate the capability of Akt-MSCs or media collected from these cells to stimulate EPC migration and tube formation in vitro and the ability to enhance neovascularization in the post infarcted heart in vivo. We will also examine the contribution of the potential Akt-MSC derived factors towards these processes. Next we will study the role of bone marrow progenitor cells in Akt-MSC induced neovascularization in ischemic myocardium in irradiated/bone marrow transplantation models using genetically marked donor cells. Moreover we will use a "suicide" gene delivery approache to selectively eliminate the bone marrow progenitor cells which may play a role in Akt-MSC mediated neovascularization. Finally, we will investigate the role of hypoxia and Akt regulated transcription factor HIF11 as a molecular switch which may regulate Akt-MSC induced bone marrow cell recruitment to the ischemic myocardium. These studies should help elucidate the mechanism by which Akt-MSCs provide such dramatic and long term protection of the infarcted myocardium and may identify potential therapeutic approaches. PUBLIC HEALTH RELEVANCE The protective effects of adult bone marrow stem cells in the injured heart have been demonstrated, however, the exact mechanism is unknown. In this study, we will examine the effects of these bone marrow cells in the development of new blood vessels in the infarcted heart. Enhancement of new vessels in the heart will greatly aid in the treatment of coronary vessel disease and heart failure.

描述（由申请人提供）：我们以前已经证明，间充质干细胞（MSC），当注射到心肌梗死（MI）后的啮齿动物心脏，增强心肌修复和恢复心脏功能。此外，这种保护作用通过用细胞保护基因Akt（Akt-MSC）转导MSC而增强。此外，我们的数据表明，这些细胞表达旁分泌介质，减少心肌损伤。在本申请中，我们假设Akt-MSC增强心肌修复，部分是通过其对血管生成的旁分泌作用。事实上，我们在实验室中已经证明Akt-MSC表达多种血管生成细胞因子，例如VEGF和FGF。此外，从这些细胞收集的培养基在体外诱导内皮细胞迁移和管形成。重要的是，我们最近已经证明，Akt-MSC诱导梗死心脏中的新血管形成。由于已经显示血管生成信号动员骨髓来源的内皮祖细胞（EPC），其归巢到缺血心肌，我们假设（i）Akt-MSC通过旁分泌机制增加梗死后心脏中的血管生成，并有助于改善心肌修复和功能;（ii）Akt-MSC部分通过募集骨髓来源的EPCs增加血管生成;和（iii）Akt-MSC部分通过募集骨髓来源的EPCs增加血管生成。MSC介导的EPC募集由HIF 11控制。为了验证这些假设，我们将研究Akt-MSC或从这些细胞收集的培养基在体外刺激EPC迁移和管形成的能力以及在体内增强梗死后心脏中的新血管形成的能力。我们还将研究潜在的Akt-MSC衍生因子对这些过程的贡献。接下来，我们将使用遗传标记的供体细胞研究骨髓祖细胞在辐照/骨髓移植模型中在缺血心肌中Akt-MSC诱导的新血管形成中的作用。此外，我们将使用“自杀”基因递送方法选择性地清除可能在Akt-MSC介导的新生血管形成中起作用的骨髓祖细胞。最后，我们将研究缺氧和Akt调节的转录因子HIF 11作为分子开关的作用，其可能调节Akt-MSC诱导的骨髓细胞向缺血心肌的募集。这些研究有助于阐明Akt-MSCs对梗死心肌提供如此显著和长期保护的机制，并可能确定潜在的治疗方法。 成人骨髓干细胞对受损心脏的保护作用已被证实，但确切的机制尚不清楚。 在这项研究中，我们将检查这些骨髓细胞在梗死心脏新血管发育中的作用。 心脏中新血管的增强将极大地有助于冠状血管疾病和心力衰竭的治疗。