Human CDX2 Cells and Cardiac Repair

项目摘要 已知胎儿来源的胎盘细胞在怀孕期间进入母体循环，并可能持续存在 母体组织几十年来一直是微嵌合体。我们已经报道了胎儿细胞选择性地回家受伤 并分化为不同的心脏谱系。使用增强型绿色 荧光蛋白(EGFP)标记的胎儿，我们展示了移植和心脏分化 母体心脏损伤区域的多能胎儿细胞。在体外，从母体心脏中分离出的胎儿细胞 重述这些分化途径，形成血管管并自发跳动 心肌细胞以融合非依赖的方式。在母体中有很大比例(~40%)的胎儿细胞 心脏表达尾部相关同源盒2(CDX2)，以前与滋养层干细胞相关。 利用cre-lox技术进行谱系追踪，我们现在已经证明，cdx2细胞可以从 在体外可形成搏动的心肌细胞和血管细胞。此外，他们还 表现出转录特征，表明有能力逃避宿主免疫监视。蛋白质组学 与ES细胞相比，对这些细胞的研究显示出明显的生长、存活和归巢优势，但 保留胚胎干细胞的“干性”特性。因此，转录和蛋白质组学分析揭示了 可以帮助开发异基因细胞治疗方法的理想品质。作为进一步的支持 在这一目标中，我们已经证明了cdx2细胞强烈且特异地定位于梗塞的心脏。 注射到尾静脉后，体内分化为心肌细胞和血管。磁共振成像 表现出显著和持续的收缩能力增强(美国国家科学院院刊手稿修订)。我们 也表明CDX2细胞可以从人足月胎盘中分离出来。我们的最终目标是翻译 这些研究用于临床，我们提出三个目标，利用最尖端的技术在 科学才能实现这一点。在目标1中，我们将发现人类CDX2细胞独特的细胞表面标记 通过转录组/蛋白质组图谱，并探索它们的归巢机制。在《目标2》中，我们试图理解 这些细胞的免疫学特性，以帮助开发同种异体人类细胞疗法。 我们还将确定他们形成畸胎瘤的倾向。在目标3中，我们将确认CDX2细胞也提供 心肌梗死后在体内外向有功能的心肌细胞和血管细胞分化 在野生型和NOD/SCID小鼠中诱导。我们还将测试这些细胞是否能增强心肌 心梗后的功能，因为这将有助于我们设计治疗策略。

Project Summary Fetal-derived placenta cells are known to enter the maternal circulation during pregnancy and may persist in maternal tissue for decades as microchimeras. We have reported that fetal cells selectively home to injured maternal myocardium and undergo differentiation into diverse cardiac lineages. Using enhanced green fluorescent protein (eGFP)-tagged fetuses, we demonstrated engraftment and cardiac differentiation of mulitpotent fetal cells in injury zones of maternal hearts. In vitro, fetal cells isolated from maternal hearts recapitulate these differentiation pathways, forming vascular tubes and spontaneously beating cardiomyocytes in a fusion-independent manner. A significant proportion (~40%) of fetal cells in maternal hearts express Caudal-related homeobox2 (Cdx2), previously associated with trophoblast stem cells. Utilizing cre-lox technology for lineage-tracing, we have now shown that Cdx2 cells can be isolated from end-gestation placenta and can form beating cardiomyocytes and vascular cells in vitro. Furthermore, they exhibit a transcriptomic signature that suggests an ability to evade host immune surveillance. Proteomic studies of these cells compared to ES cells reveal distinct growth, survival and homing advantages, but with retention of the `stemness' properties of ES cells. Thus the transcriptomic and proteomic analysis reveal desirable qualities that can aid in the development of an allogeneic cell therapy approach. In further support of this objective, we have demonstrated that Cdx2 cells home robustly and specifically to infarcted hearts upon injection into the tail vein, with differentiation in vivo to cardiomyocytes and blood vessels. MRI demonstrates significant and sustained enhancement of contractility (manuscript in revision at PNAS). We have also shown that CDX2 cells can be isolated from human term placentas. Our final goal is to translate these studies for clinical use, and we propose three aims utilizing the most cutting-edge technologies in science in order to achieve this. In aim 1, we will uncover unique cell surface markers of human CDX2 cells via trancriptome/proteome profiling and explore their homing mechanisms. In aim 2, we seek to understand immunologic properties of these cells in order to aid in the development of allogeneic human cell therapy. We will also ascertain their propensity to form teratomas. In aim 3, we will confirm that CDX2 cells also give rise to functional cardiomyocytes and vascular cells in vitro and in vivo after myocardial infarction (MI) is induced in wild-type and NOD/SCID mice. We will also test whether these cells enhance myocardial function after MI, as this will aid us in designing a therapeutic strategy.