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

• 批准号: 7882861
• 负责人: CHRISTOPHER NIYIBIZI
• 金额: $ 20.94万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 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/7882861
• 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; Natural regeneration; 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; transcription factor

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）：本申请是一份探索性R21资助申请，响应NIH探索性/发展性研究资助计划（家长R21）： PA-09-164。本探索性应用的目的是评估诱导多能干细胞（iPSC）在全身移植后向骨组织中的迁移、植入和分化。在全身注射后，祖细胞迁移到骨组织的能力对于治疗广泛性骨骼疾病至关重要。在动物模型中评估MSCs治疗成骨不全的可移植性的研究产生了不同的结果。应用间充质干细胞治疗成骨不全的大多数争议可能是由于不同研究者用于移植的细胞类型。胚胎干细胞（ESC）来源于囊胚的内部细胞群，可以产生身体的任何细胞类型，并且可以在不失去其多能性的情况下无限扩增，因此它们在包括广泛性骨骼疾病在内的各种疾病的细胞治疗中具有更大的应用潜力。然而，由于伦理问题，在利用这些细胞的力量方面几乎没有取得进展。最近，研究表明，通过引入四种转录因子的组合，小鼠和人类成纤维细胞可以被重编程为esc样状态；Oct-3/4, Sox2， c-Myc和Klf4。被称为诱导多能干细胞（iPSC）的重编程细胞为产生用于治疗目的和药物筛选的患者特异性干细胞提供了机会。作为了解iPSC在广泛性骨骼疾病治疗中的未来应用的前奏曲，本探索性应用建议评估iPSC衍生的间充质干细胞移植到成骨不全小鼠模型后的迁移、植入和分化。为实现上述任务，将采用以下目标：1)通过重编程小鼠尾尖成纤维细胞产生的iPSC生成间充质干细胞2)评估iPSC衍生的MSC样细胞在成骨不全小鼠模型骨组织中的可移植性、迁移性、植入性和分化性。初步数据表明，我们可以通过对小鼠尾尖成纤维细胞进行重编程来生成iPSC，并且这些细胞表现出类似ESC的状态。我们将培育携带胶原蛋白突变的杂合小鼠，产生3种小鼠基因型（野生型、杂合型和纯合型）。从野生型小鼠中制备用于重编程的成纤维细胞，并将同基因杂合和纯合小鼠作为细胞受体。初步数据表明，iPSC短暂暴露于TGF-21会产生具有间充质干细胞特征的细胞。我们将使用这种方法来产生用于移植的细胞。将通过生物成像、组织学和基因表达分析来评估细胞在体内的迁移、移植和分化情况。这一探索性应用的结果将为未来研究iPSC细胞在肌肉骨骼组织修复和再生中的应用提供一个平台。