Derivation and Transplantation of Histocompatible Pluripotent Stem Cells

组织相容性多能干细胞的衍生和移植

• 批准号: 7924653
• 负责人: David W Russell
• 金额: $ 31.2万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7924653
• 关键词: Affect; Allogenic; American; Autologous; Behavior; CD34 gene; Cell Line; Cell Therapy; Cell Transplants; Cells; Chimeric Proteins; Chromosomes, Human, Pair 6; Clinic; Clinical; Clinical Protocols; Clinical Trials; Dependovirus; Derivation procedure; Development; Developmental Gene; Embryo; Engineering; Engraftment; Ensure; Fibroblasts; Ganciclovir; Gene Expression; Gene Expression Profiling; Gene Targeting; Genes; Genome; Globin; Goals; HLA-A gene; Haplotypes; Hematopoietic; Histocompatibility Antigens; Histocompatibility Testing; Human; Human Chromosomes; Immunosuppression; In Vitro; Individual; Investments; Karyotype determination procedure; Long Terminal Repeats; Macaca; Macaca nemestrina; Mesenchymal Stem Cells; Mitotic; Mitotic Recombination; Modeling; Mus; Patients; Phenotype; Pluripotent Stem Cells; Population; Preclinical Testing; Problem Solving; Procedures; Proliferating; Recombinants; Regenerative Medicine; Resistance; Siblings; Site; Solutions; Source; Spumavirus; Stem cells; Teratoma; Testing; Therapeutic; Time; Transgenes; Transplantation; United States; arm; blastomere structure; cell type; erythroid differentiation; human disease; human embryonic stem cell; hygromycin A; improved; in vivo; induced pluripotent stem cell; nonhuman primate; recombinase; research study; stem cell therapy; telomere; therapeutic cloning; vector

Human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) can only be used effectively in recipients with the same (or nearly the same) histocompatibility type, to avoid rejection or toxic immunosuppression. To overcome this, various strategies have been proposed to derive patient-specific stem cells. These often involve controversial procedures such as therapeutic cloning, and they require a major investment in time and money for every patient that is impractical for routine clinical use. The goal of this proposal is to solve this problem and create a panel of pluripotent stem cells that is histocompatible with a significant percentage of the U.S. population. This panel will consist of cells that are homozygous at the HLA locus, so only one set of major histocompatibility antigens is expressed. By choosing the most common haplotypes, 5-10 HLA-homozygous lines should match over 20-25% of Americans at HLA-A, B and DR loci, and 50 lines should match over 70% of the population. To accomplish this, we will derive a set of iPSCs with common HLA haplotypes from different types of non-embryonic cells. Integrating vectors will be used to deliver reprogramming transgenes, then they will be removed from the cells to create transgene-free iPSCs. Adeno-associated virus (AAV) gene targeting vectors will be used to insert a plus/minus selectable marker next to the HLA locus on human chromosome 6, then mitotic recombinants will be isolated that lost the marker gene and converted the short arm of chromosome 6 to homozygosity, including the entire HLA locus (an approach that has already succeeded in our pilot experiments). The parental iPSCs and their HLA-homozygous derivatives will be characterized extensively by SNP-chip analysis, karyotyping, and differentiation potential. This project will establish a bank of "patient-specific" HLA/MHC-homozygous stem cells that can be used for all types of regenerative medicine, avoiding the problems of graft rejection and immunosuppression, with significant potential for improving the health of Americans.

人类胚胎干细胞（ESC）和诱导多能干细胞（IPSC） 只能有效地用于具有相同（或几乎相同）的接受者 组织相容性类型，以避免排斥或有毒免疫抑制。克服 已经提出了各种策略来推导患者特异性干细胞。这些 通常涉及有争议的程序，例如治疗克隆，它们需要一个 每位不切实际的患者对时间和金钱的重大投资 临床用途。该建议的目的是解决此问题并创建一个面板 组织兼容的多能干细胞与美国相同的比例很大 人口。该面板将由HLA基因座的纯合细胞组成，因此 仅表达一组主要的组织相容性抗原。通过选择最多 常见的单倍型，5-10 HLA-HOMOZYGOUS线应匹配20-25％ HLA-A，B和Loci博士的美国人和50行应匹配70％以上 人口。为此，我们将带有常见HLA的一组IPSC 来自不同类型的非胚胎细胞的单倍型。将使用集成向量 要提供重编程转基因，然后将其从单元格中删除以创建 无基因IPSC。将使用腺体相关病毒（AAV）基因靶向载体 插入HLA基因座旁边的Plus/减去可选标记的人类染色体 6，然后将分离有丝分裂重组剂，以失去标记基因并转换 染色体6至纯合性的短臂，包括整个HLA基因座（一个 在我们的试点实验中已经成功的方法）。父母ipscs和 他们的HLA-HOMOZYGOUS衍生物将通过SNP-Chip广泛表征 分析，核分型和分化潜力。该项目将建立一家银行 可用于所有类型 再生医学，避免了移植排斥和免疫抑制的问题， 具有巨大的潜力来改善美国人的健康状况。