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）：本申请是响应 NIH 探索性/发展研究资助计划（母 R21）的探索性 R21 资助申请：PA-09-164。本探索性应用的目标是评估诱导多能干细胞（iPSC）来源的细胞在全身移植后向骨骼组织的迁移、植入和分化。全身注射后祖细胞迁移至骨骼组织的能力对于全身骨骼疾病的治疗至关重要。评估间充质干细胞移植治疗成骨不全症的动物模型研究产生了不同的结果。应用间充质干细胞治疗成骨不全症的大部分争议可能源于不同研究者用于移植的细胞类型。源自囊胚内细胞团的胚胎干细胞（ESC）可以产生身体的任何细胞类型，并且可以无限增殖而不失去其多能性，因此在细胞治疗各种疾病方面具有更大的应用潜力，包括全身骨骼疾病。然而，出于伦理方面的考虑，在利用这些细胞的力量方面几乎没有取得任何进展。最近，已经证明，通过引入四种转录因子的组合，可以将小鼠和人类成纤维细胞重编程为ESC样状态； 10 月 3 日，Sox2，c-Myc 和 Klf4。被称为诱导多能干细胞 (iPSC) 的重编程细胞为生成患者特异性干细胞提供了机会，用于治疗目的和药物筛选。作为了解 iPSC 在全身骨骼疾病治疗中的未来应用的前奏，目前的探索性应用建议评估 iPSC 衍生的 MSC 在移植到成骨不全小鼠模型中后的迁移、植入和分化。为实现上述任务，将采取以下目标： 1) 从通过重编程小鼠尾尖成纤维细胞产生的 iPSC 生成 MSC 2) 评估源自 iPSC 的 MSC 样细胞进入成骨不全症小鼠模型骨骼组织的可移植性、迁移、植入和分化。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产生 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，以产生 3 种小鼠基因型（野生型、杂合型和纯合型）。用于重编程的成纤维细胞将从野生型小鼠中制备，同基因杂合和纯合小鼠将作为细胞受体。初步数据表明，iPSC 短暂暴露于 TGF-21 会产生表现出 MSC 特征的细胞。我们将使用这种方法来生成用于移植的细胞。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、植入和分化。这一探索性应用的结果将为未来 iPSC 细胞在肌肉骨骼组织修复和再生中的应用研究提供一个平台。公共健康相关性：本探索性应用的目标是评估诱导多能干细胞 (iPSC) 来源的细胞在全身移植后向骨骼组织的迁移、植入和分化。这一探索性应用的结果将为 iPSC 细胞在肌肉骨骼组织修复和再生中的未来研究提供平台