Endodermal progenitor cell characterization

内胚层祖细胞表征

• 批准号: 8334475
• 负责人: PAUL J GADUE
• 金额: $ 33.5万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-20 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8334475
• 关键词: Address; Albumins; Biological Assay; Biology; Cell Fate Control; Cell Line; Cell Maintenance; Cell Therapy; Cell physiology; Cells; Data; Development; Diabetic mouse; Disease; Ectoderm; Embryo; Embryonic Development; Endoderm; Endoderm Cell; Gene Expression Microarray Analysis; Generations; Genes; Genetic; Germ Layers; Glucose; Hepatocyte; Histocompatibility Testing; Human; Immune; In Vitro; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Lead; Liver; Liver diseases; Lung; Maintenance; Mesoderm; Modeling; Mus; Organ; Pancreas; Pathway interactions; Patients; Population; Process; Protocols documentation; RNA Interference; Replacement Therapy; Risk; Role; Running; Safety; Signal Pathway; Signal Transduction; Solutions; Source; Staging; Stem cells; Study models; System; Technology; Teratoma; Testing; Tissues; To specify; Transplantation; Transplanted tissue; Tube; Undifferentiated; Work; Xeno; cell type; chemical genetics; embryonic stem cell; gastrointestinal; human embryonic stem cell; in vitro Assay; in vivo; induced pluripotent stem cell; insulin secretion; islet; notch protein; progenitor; self-renewal; small molecule; stem; stem cell biology; stem cell population; transcription factor; tumor

DESCRIPTION (provided by applicant): Stem cell biology offers tremendous potential to both study and model disease in vitro as well as in the treatment of disease. Diseases of endodermal organs including type I diabetes and various liver diseases are of great concern. There are currently not sufficient donors for islet cells or livers necessary to treat the number of patients requiring transplants. The use of stem cell populations to generate pancreatic islet cells and liver would offer a potential solution to this problem. Currently, work with human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells offer hope in the treatment of these diseases. Both of these cell types can be expanded in culture and have the potential to differentiate into any cell type in the body. Unfortunately, there are several hurdles that need to be overcome before cell replacement therapy becomes a reality. First, both ES and iPS cells when transplanted directly can form tumors. Therefore, it is of critical importance to transplant pure differentiated cell types. In addition, the generation of mature function cell types from these early stem cell populations has proved difficult. To attempt to address these concerns we propose to generate endodermal progenitor cell (EP) lines. EP cells, like ES and iPS cells, can be expanded in culture but lack tumor forming potential. EP cells can also differentiate into endodermal cell types such as hepatocytes and pancreatic cells. Preliminary data suggests that we have found the culture conditions to generate EP cells from human ES cells. We propose to further characterize EP cells, generate EP cells from multiple human ES and iPS cell lines and to differentiate them into hepatocytes and pancreatic islet cells. These differentiated progeny will be functionally assayed in a variety of in vitro and in vivo systems to determine if both ES cell and iPS cell derived EP cell lines are functionally equivalent. Finally, we propose to investigate the mechanisms that control EP cell generation and maintenance. Preliminary data indicates that Notch signaling may promote EP cell formation. We will expand upon these findings, examining Notch signaling utilizing both genetic systems and small molecules to activate or repress the Notch pathway and assay the effects on EP cell formation and maintenance. In addition, other possible candidates discovered using gene expression microarray analysis will also be assayed by RNAi knockdown in EP cells. Genes found to be required in EP cells will then be studied in the context of Notch signaling to establish a signaling hierarchy for EP cell formation and maintenance. This information may make generating EP cells more efficient and may lead to a better understanding of the mechanisms controlling stem cell populations in general.

描述（由申请人提供）：干细胞生物学为体外研究和模型疾病以及疾病治疗提供了巨大的潜力。内胚层器官疾病，包括I型糖尿病和各种肝脏疾病，备受关注。目前没有足够的胰岛细胞或肝脏捐献者来治疗需要移植的患者数量。使用干细胞群产生胰岛细胞和肝脏将为这个问题提供潜在的解决方案。目前，人类胚胎干（ES）细胞和诱导多能干（iPS）细胞的研究为治疗这些疾病带来了希望。这两种细胞类型都可以在培养物中扩增，并且有可能分化成体内的任何细胞类型。不幸的是，在细胞替代疗法成为现实之前，还需要克服一些障碍。首先，ES细胞和iPS细胞直接移植时都可以形成肿瘤。因此，移植纯分化细胞类型至关重要。此外，事实证明，从这些早期干细胞群产生成熟功能细胞类型是很困难的。为了尝试解决这些问题，我们建议生成内胚层祖细胞（EP）系。 EP 细胞与 ES 和 iPS 细胞一样，可以在培养物中扩增，但缺乏肿瘤形成潜力。 EP细胞还可以分化为内胚层细胞类型，例如肝细胞和胰腺细胞。初步数据表明，我们已经找到了从人 ES 细胞产生 EP 细胞的培养条件。我们建议进一步表征 EP 细胞，从多种人类 ES 和 iPS 细胞系中产生 EP 细胞，并将其分化为肝细胞和胰岛细胞。这些分化的后代将在各种体外和体内系统中进行功能分析，以确定 ES 细胞和 iPS 细胞衍生的 EP 细胞系在功能上是否等效。最后，我们建议研究控制 EP 细胞生成和维持的机制。初步数据表明Notch信号传导可能促进EP细胞形成。我们将扩展这些发现，利用遗传系统和小分子来检查 Notch 信号传导，以激活或抑制 Notch 通路，并分析对 EP 细胞形成和维持的影响。此外，使用基因表达微阵列分析发现的其他可能的候选者也将通过 EP 细胞中的 RNAi 敲低进行分析。然后将在 Notch 信号传导背景下研究 EP 细胞所需的基因，以建立 EP 细胞形成和维持的信号传导层次结构。这些信息可能会使 EP 细胞的产生更加有效，并可能导致更好地理解控制干细胞群的一般机制。