Transplantability of induced pluripotent stem cells for skeletal tissues

诱导多能干细胞用于骨骼组织的可移植性

基本信息

批准号：
8037778
负责人：
CHRISTOPHER NIYIBIZI
金额：
$ 16.75万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2012-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8037778
关键词：
Adult Aging Animal Model Applications Grants Biological Models Breeding Cell Differentiation process Cell Therapy Cell Transplantation Cells Characteristics Clinical Trials Collagen Data Development Disease Engraftment Ethics Exhibits Fibroblasts Future Gene Expression Genotype Goals Harvest Histology Human Injection of therapeutic agent Inner Cell Mass Investigation Manuscripts Mesenchymal Mesenchymal Stem Cells Mus Musculoskeletal Mutation Osteogenesis Imperfecta Parents Patients Preclinical Drug Evaluation Protocols documentation Research Personnel Research Project Grants Stem cell transplant Stem cells Stromal Cells Tail Therapeutic Transplantation Tretinoin United States National Institutes of Health Veins Viral Vector Work Wound Healing bioimaging blastocyst c-myc Genes cell type embryonic stem cell in vivo induced pluripotent stem cell migration mouse model pluripotency progenitor programs public health relevance response skeletal disorder skeletal tissue tissue regeneration transcription factor

项目摘要

DESCRIPTION (provided by applicant): The present application is an exploratory R21 grant application in response to NIH Exploratory/ Developmental Research Grant Program (Parent R21): PA-09-164. The goals of the present exploratory application are to assess migration, engraftment and differentiation of cells derived from induced pluripotent stem cells (iPSC) into skeletal tissues following systemic transplantation. The ability of progenitors to migrate to skeletal tissues following systemic injection is critical for the treatment of generalized skeletal diseases. Studies in animal models assessing transplantability of MSCs to treat osteogenesis imperfecta have generated mixed results. Most of the controversies in the application of MSCs to treat OI may be due to the types of cells used for transplantation by different investigators. Embryonic stem cells (ESC) derived from the inner cell mass of the blastocyst can give rise to any cell type of the body and can be expanded indefinitely without losing their pluripotency, thus these possess a greater potential for application in cell therapies for various diseases including generalized skeletal diseases. Because of ethical concerns however, little progress has been made in harnessing the power of these cells. Recently, it has been demonstrated that mouse and human fibroblasts can be reprogrammed into an ESC-like state by introducing combinations of four transcription factors; Oct-3/4, Sox2, c-Myc and Klf4. The reprogrammed cells referred to as induced pluripotent stem cells (iPSC) offer opportunities for generating patient specific stem cells for therapeutic purposes and drug screening. As a prelude to understanding the future application of iPSC for generalized skeletal disease treatment, the present exploratory application proposes to assess migration, engraftment and differentiation of MSCs derived from iPSC following transplantation into a mouse model of osteogenesis imperfecta. The following aims will be employed to achieve the above tasks; 1) generate MSCs from iPSC created by reprogramming mouse tail tip fibroblasts 2) Assess transplantability, migration, engraftment and differentiation of MSC like cells derived from iPSCs into the skeletal tissues of a mouse model of OI. Preliminary data show that we can generate iPSC by reprogramming murine tail tip fibroblasts and that the cells exhibit ESC like state. We will breed heterozygous mice that carry a collagen mutation to generate 3 mice genotypes (wildtype, heterozygous and homozygous). Fibroblasts for reprogramming will be prepared from wildtype mice and the syngeneic heterozygous and homozygous mice will be the cell recipients. Preliminary data indicate that brief exposure of iPSC to TGF-21 generates cells that exhibit MSCs characteristics. We will use this approach to generate cells for transplantation. Migration, engraftment and differentiation of the cells in vivo will be assessed by bioimaging, histology and gene expression analysis. The results from this exploratory application will provide a platform for future investigations in the application of iPSC cells for musculoskeletal tissue repair and regeneration. PUBLIC HEALTH RELEVANCE: The goals of the present exploratory application are to assess migration, engraftment and differentiation of cells derived from induced pluripotent stem cells (iPSC) into skeletal tissues following systemic transplantation. The results from this exploratory application will provide a platform for future investigations of iPSC cells in musculoskeletal tissue repair and regeneration

描述（由申请人提供）：本申请是探索性R21授予申请，以响应NIH探索性/发展研究赠款计划（父级R21）：PA-09-164。本探索性应用的目标是评估从诱导的多能干细胞（IPSC）衍生成的细胞的迁移，植入和分化，在全身移植后将其迁移到骨骼组织中。全身注射后祖细胞迁移到骨骼组织的能力对于治疗广义骨骼疾病至关重要。在评估MSC治疗骨化不完美的动物模型中的研究产生了混合的结果。在应用MSC治疗OI中，大多数争议可能是由于不同研究者用于移植的细胞的类型。源自胚泡的内部细胞质量的胚胎干细胞（ESC）可以引起人体的任何细胞类型，并且可以无限期地扩展而不会失去其多能性，因此，这些细胞具有更大的潜力，可以在细胞疗法中用于包括广义骨骼疾病在内的各种疾病。然而，由于道德上的关注，在利用这些细胞的力量方面几乎没有取得进展。最近，已经证明，通过引入四个转录因子的组合，可以将小鼠和人成纤维细胞重编程为ESC样状态。 OCT-3/4，SOX2，C-MYC和KLF4。重编程的细胞称为诱导的多能干细胞（IPSC）为治疗目的和药物筛查提供了生成患者特定干细胞的机会。作为理解IPSC在广义骨骼疾病治疗中的未来应用的前奏，本探索性应用建议评估从IPSC移植到成骨的小鼠模型中，从IPSC中得出的MSC的迁移，植入和分化。将采用以下目标来完成上述任务； 1）通过重新编程小鼠尾尖成纤维细胞产生的IPSC的MSC 2）评估MSC的移植，迁移，移植和分化，例如从IPSC中得出的细胞，从IPSC中得出的细胞中的iPSC中的细胞进入了OI小鼠模型的骨骼组织。初步数据表明，我们可以通过重编程鼠尾尖尖成纤维细胞来生成IPSC，并且细胞表现出ESC的状态。我们将繁殖携带胶原蛋白突变的杂合小鼠，以产生3只小鼠基因型（野生型，杂合和纯合子）。用于重编程的成纤维细胞将由野生型小鼠制备，而合成生杂合子和纯合小鼠将是细胞受体。初步数据表明，短暂的IPSC暴露于TGF-21会产生具有MSCS特征的细胞。我们将使用这种方法来生成细胞进行移植。体内细胞的迁移，植入和分化将通过生物成像，组织学和基因表达分析来评估。此探索性应用的结果将为IPSC细胞用于肌肉骨骼组织修复和再生的应用提供一个平台。公共卫生相关性：当前探索性应用的目标是评估从全身移植后从诱导的多能干细胞（IPSC）衍生出的细胞迁移，植入和分化到骨骼组织。探索性应用的结果将为肌肉骨骼组织修复和再生中IPSC细胞的未来研究提供一个平台