Protein-induced human iPS cells for personalized cell therapy of PD

蛋白质诱导的人类 iPS 细胞用于 PD 个性化细胞治疗

基本信息

批准号：
8481598
负责人：
Kwang-Soo Kim
金额：
$ 32.69万
依托单位：
MCLEAN HOSPITAL
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：
2010-06-01 至 2015-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8481598
关键词：
Animal Model Behavioral Biological Biomedical Research Calculi Cell Line Cell Therapy Cell Transplantation Cells Clinical Data Clinical Research Collaborations Controlled Clinical Trials Copy Number Polymorphism Corpus striatum structure Custom DNA Data Defect Development Disease Disease model Double-Blind Method Embryo Epigenetic Process Exhibits Family Fibroblasts Functional disorder Gene Expression Generations Genes Genetic Genomics Goals Human In Vitro Infant Karyotype Laboratories Lead Lesion Letters Location Methods Midbrain structure Molecular Morphology Motor Neurodegenerative Disorders Neurons Newborn Infant Oligonucleotides Outcome Oxidopamine Parkinson Disease Pathology Patients Peptides Physiological Placebo Control Pluripotent Stem Cells Property Proteins Publishing Rattus Recovery Regimen Resolution Retroviridae Rodent Model Somatic Cell Source Southern Blotting Staging Stem Cell Research Stem cells Substantia nigra structure Techniques Testing Therapeutic Tissues Transgenes Transplantation Validation Viral Virus Work aphakia mice base c-myc Genes cell type clinical application disease mechanisms study dopaminergic neuron embryonic stem cell fetal human disease human embryonic stem cell human embryonic stem cell line improved in vivo induced pluripotent stem cell insight male nerve stem cell neurotransmission novel open label preclinical study public health relevance research study retroviral transduction stem cell technology tumor vector viral DNA

项目摘要

DESCRIPTION (provided by applicant): Parkinson's disease (PD) is the second most common neurodegenerative disorder in which the main pathology is selective degeneration of midbrain dopaminergic (mDA) neurons in the substantia nigra. Because of this specific cell loss, PD is considered to be a prime target disease for cell-based therapy. Indeed, numerous clinical and preclinical studies demonstrated the proof-of-principle that cell transplantation is a viable therapeutic regimen for PD treatment once an ideal and unlimited cell source can be established (reviewed in (Redmond, 2002; Li et al., 2008a; Lindvall and Kokaia, 2009). In 2006, Shinya Yamanaka and his colleagues published their groundbreaking work showing that pluripotent stem cells, so called "induced pluripotent stem cells (iPSCs)", can be generated from somatic cells by retroviral transduction of four reprogramming factors (i.e., Oct4, Sox2, Klf4 and c-Myc)(Takahashi and Yamanaka, 2006). Subsequent successful generation of human iPSCs by similar methods (Takahashi et al., 2007; Yu et al., 2007; Park et al., 2008a) offered the possibility to generate disease- or patient-specific stem cells without destruction of embryos. Indeed, these iPSCs offer unprecedented potentials for biomedical research, disease mechanism study, and personalized cell-based therapies. However, current iPSCs suffer from major drawbacks including multiple viral integrations and remaining transgenes at various chromosomal locations, any of which may cause unpredictable genetic dysfunction and/or tumor formation, making these cells unsuitable for clinical applications (Yamanaka, 2009a). With the long-term goal of developing a personalized cell-based therapy of PD using iPSCs, we propose the following three specific aims. First, based on our preliminary results (Kim et al., 2009a), we will establish iPSC lines from healthy and sporadic PD subjects by a novel, DNA-free reprogramming method (i.e., direct delivery of reprogramming proteins). We will extensively evaluate morphological, gene expression, epigenetic, and in vitro and in vivo differentiation properties to establish iPSC lines exhibiting properties similar to human embryonic stem cells (hESCs). We will compare their properties with those of hESC lines and iPSCs generated by conventional retroviral methods. In addtion, we will investigate their chromosomal integrity by state of the art copy number variation analysis. Second, once authentic iPSC lines are established, we will fully characterize and compare their in vitro differentiation properties into neural progenitors and midbrain DA neurons. We will optimize their differentiation into A9 DA neurons and evaluate the molecular, cellular, and electrophysiological characters of DA neurons from these iPS cells. Third, we will initiate to evaluate the potential functional benefits of these iPSC-derived DA neurons in two rodent models of PD; aphakia mice and 6-OHDA lesioned rats, both of which are well established in our laboratory. Biological and behavioral outcomes of these transplantation studies will be systematically investigated. Overall, our proposed experiments will provide invaluable insights and stepping stones, leading to the development of safe, realistic, and ideal cell source for personalized cell-based therapy of PD.

描述（由申请人提供）：帕金森氏病（PD）是第二个最常见的神经退行性疾病，其中主要病理是尼格拉岛中脑多巴胺能（MDA）神经元的选择性变性。由于这种特定的细胞损失，PD被认为是基于细胞治疗的主要靶向疾病。 Indeed, numerous clinical and preclinical studies demonstrated the proof-of-principle that cell transplantation is a viable therapeutic regimen for PD treatment once an ideal and unlimited cell source can be established (reviewed in (Redmond, 2002; Li et al., 2008a; Lindvall and Kokaia, 2009). In 2006, Shinya Yamanaka and his colleagues published their groundbreaking work showing that pluripotent所谓的“诱导多能干细胞（IPSC）”可以通过逆转录病毒转导四个重编程因子（即OCT4，SOX2，KLF4和C-MYC）从体细胞中产生的干细胞（Takahashi和Yamanaka，Yamanaka，2006年）。 2007年;不可预测的遗传功能障碍和/或肿瘤形成，使这些细胞不适合临床应用（Yamanaka，2009a）。长期目标是使用IPSC开发基于个性化细胞的PD疗法，我们提出了以下三个特定目标。首先，基于我们的初步结果（Kim等，2009a），我们将通过一种新型的无DNA重新编程方法（即直接递送重编程蛋白）从健康和零星的PD受试者中建立IPSC线。我们将广泛评估形态学，基因表达，表观遗传学以及体外和体内分化特性，以建立具有类似人类胚胎干细胞（HESC）的特性的IPSC线。我们将将它们的属性与常规逆转录病毒方法生成的hESC线和IPSC的属性进行比较。另外，我们将根据最新拷贝数变化分析的状态研究它们的染色体完整性。其次，一旦建立了正宗的IPSC线，我们将充分表征并将其体外分化特性与神经祖细胞和中脑DA神经元进行比较。我们将优化它们为A9 DA神经元的分化，并评估来自这些IPS细胞的DA神经元的分子，细胞和电生理特征。第三，我们将开始评估在PD的两个啮齿动物模型中这些IPSC衍生的DA神经元的潜在功能益处。 Aphakia小鼠和6-OHDA病变的大鼠在我们的实验室中都建立了良好的大鼠。这些移植研究的生物学和行为结果将被系统地研究。总体而言，我们提出的实验将提供宝贵的见解和垫脚石，从而开发安全，现实且理想的细胞来源，用于基于PD的个性化细胞治疗。