Embryonic Stem Cell Derived HSC Transplantation in Rhesus Macaque Model

恒河猴模型中胚胎干细胞衍生的 HSC 移植

• 批准号: 7314064
• 负责人: PEIMAN HEMATTI
• 金额: $ 13.64万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-14 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7314064
• 关键词: Adult; Animal Model; Bone Marrow; CD34 gene; Cell Differentiation process; Cell Line; Cell Therapy; Cells; Chimerism; Clinical; Coculture Techniques; Developmental Biology; Disease; ES Cell Line; Embryo; Engraftment; Gene Proteins; Gene Transfer; Gene-Modified; Generations; Genes; Goals; Hematopoiesis; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Human; Immunodeficient Mouse; In Vitro; Macaca mulatta; Malignant - descriptor; Mesenchymal; Methodology; Modeling; Mus; Non-Malignant; Numbers; Patients; Play; Population; Pre-Clinical Model; Primates; Principal Investigator; Process; Proteins; Recombinants; Research; Research Personnel; Role; SCID Mice; Safety; Siblings; Source; Stromal Cells; Testing; Therapeutic; Tissues; Transplantation; Umbilical Cord Blood; Undifferentiated; abstracting; clinical application; design; efficacy evaluation; embryonic stem cell; human embryonic stem cell; improved; in vitro Assay; in vivo; mouse model; nonhuman primate; novel; peripheral blood; pre-clinical; programs; reconstitution; research study; transcription factor

DESCRIPTION (provided by applicant): Although adult hematopoietic stem cells (HSCs) are being used routinely in clinical transplantation, the lack of a suitable source of donor cells remains a major limiting factor depriving many patients of this potentially curative treatment. The ultimate goal of the Principal Investigator is to generate HSCs from embryonic stem cells (ESCs) as a novel source of cells suitable for clinical transplantation. We are proposing an intensive research program to study the developmental biology of hematopoiesis from rhesus ESCs, and to establish the rhesus macaque model as a relevant pre-clinical large animal model for the evaluation of the efficacy and safety of ESCderived HSCs. Using methodologies such as co-culturing with mesenchymal stromal cells (MSC) of murine bone marrow origin, non-human primate and human ESCs have been differentiated into primitive hematopoietic cells as defined by surrogate in vitro assays. However, those derived cells have been shown to possess only limited in vivo engraftment potential upon transplantation into NOD/SCID immunodeficient mice. Our underlying hypothesis is that by providing the appropriate extrinsic and intrinsic inductive mechanisms that play critical roles in embryonic hematopoiesis, we can effectively recapitulate those processes in vitro and thus promote the efficient differentiation of rhesus ESCs into HSCs that will be capable of engraftment and long-term hematopoietic reconstitution in vivo. In aim-1 we will generate MSC cells from rhesus ESCs and will test their potential as a hematopoietic supportive microenvironment in coculture experiments for the generation of HSCs from rhesus ESCs. In aim-2 we will test our hypothesis that the over-expression of HOXB4, a transcription factor which plays a key role in embryonic hematopoiesis, in rhesus ESCs via gene transfer, or the exogenous delivery of the HOXB4 protein, can improve the efficiency of their differentiation into HSCs. In aim-3 we will assess the in vivo engraftment potential of hematopoietic cells derived from rhesus ESCs alone or when co-transplanted with rhesus ESC-derived MSCs into NOD/SCID mice as we hypothesize that the co-transplantation of rhesus ESC-derived MSCs would potentially provide an appropriate niche and thus enhance the in vivo engraftment potential of rhesus ESCderived HSCs. The results of these proposed studies would help us to design a rational approach toward moving such cells into actual rhesus transplantation experiments. Embryonic stem cells have the potential to generate an unlimited number of diverse differentiated cells for human therapeutic applications including hematopoietic stem cells. We are proposing to develop a rhesus monkey transplantation model to study the efficacy and safety of such cells as they should be carefully tested in appropriate pre-clinical animal models before being moved into human clinical applications. (End of Abstract)

描述（由申请人提供）： 尽管成年造血干细胞（HSC）通常用于临床移植中，但缺乏合适的供体细胞来源仍然是剥夺许多潜在治疗治疗的主要限制因素。主要研究者的最终目标是从胚胎干细胞（ESC）产生HSC，作为适合临床移植的新细胞来源。我们正在提出一项密集的研究计划，以研究来自恒河猴的造血生物学的发育生物学，并建立恒河猴模型，作为一种相关的临床前大型动物模型，以评估逃脱的HSC的效果和安全性。使用诸如鼠骨髓起源的间充质基质细胞（MSC）之类的方法论，非人类灵长类动物和人类ESC已被分化为原始造血细胞，这些细胞在体外测定中定义为原始的造血细胞。然而，这些细胞已被证明只有在移植到点头/SCID免疫缺陷小鼠中时具有有限的体内植入潜力。我们的基本假设是，通过提供在胚胎造血中起着关键作用的适当外在和内在的感应机制，我们可以在体外有效地概括这些过程，从而促进将HSC的有效分化为HSC，从而有能力启发和长期血液的血液造成血液结构。在AIM-1中，我们将从恒河猴ESC产生MSC细胞，并将在共培养实验中作为造血支撑性微环境测试其潜力，从而从Rhesus Esc产生HSC。在AIM-2中，我们将检验我们的假设，即HOXB4的过表达是一种在胚胎造血中起关键作用的转录因子，通过基因转移或HOXB4蛋白的外源递送在Hoxb4蛋白中起关键作用，可以提高其分化为HSC的效率。在AIM-3中，我们将评估源自恒河猴的造血细胞的体内植入潜力，或者当与恒河猴Esc衍生的MSC共同转移到点头/SCID小鼠中时，我们假设我们假设我们的恒河所的植入率会提供较高的MSC的潜在，从而可以增强eScruse and confers consect and confers consective and consect and consect in consective nichice nichice nichiche nichiche nichiche nichiche nichice nichice tospective nichice的潜在。 HSC。这些提出的研究的结果将有助于我们设计一种将这些细胞转移到实际恒河体移植实验中的合理方法。胚胎干细胞有可能为包括造血干细胞在内的人类治疗应用生成无限数量的不同分化细胞。我们建议开发恒河猴移植模型，以研究这些细胞的疗效和安全性，因为它们应在适当的临床前动物模型中仔细测试，然后再将其转移到人类的临床应用中。 （抽象的结尾）