Liver regeneration with stems cells of uniparental origin

单亲来源干细胞的肝脏再生

• 批准号: 7779386
• 负责人: Kenneth J McLaughlin
• 金额: $ 10.36万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7779386
• 关键词: Adult; Alleles; Autologous; Cells; Chimera organism; Derivation procedure; Development; Diploidy; Disease; Donor person; ES Cell Line; Embryo; Engraftment; Fetal Liver; Fumarylacetoacetase; Generations; Genetic; Genome; Genomic Imprinting; Germ Cells; Hematopoiesis; Hematopoietic System; Hematopoietic stem cells; Hepatic; Hepatic Tissue; Hepatocyte; Hereditary Disease; Human; Hybrid Resistance; Hybrids; Immune; In Vitro; Lesion; Liver; Liver Regeneration; Liver diseases; Measures; Modeling; Mouse Strains; Mus; Mutation; Natural regeneration; Oocytes; Organ Donor; Parents; Pathology; Patients; Protein C Inhibitor; Safety; Siblings; Source; Stem cells; Therapeutic; Tissue Transplantation; Tissues; Transgenic Mice; Transplantation; alpha 1-Antitrypsin Deficiency; base; early embryonic stage; embryonic stem cell; fetal; in vivo; liver function; liver transplantation; mouse model; mutant; neonate; offspring; progenitor; reconstitution; repaired; research study; somatic cell nuclear transfer; sperm cell; stem

DESCRIPTION (provided by applicant): For many liver diseases, including those of inborn genetic origin, transplantation of donor organs is currently the only curative option. The derivation of cellular liver transplants from embryonic stem (ES) cells as an alternative source of tissue would need to be both autologous and free of the genetic disease, requirements that are not easily surmountable and also ethically controversial with ES cells derived from potentially viable fertilized or patient-specific somatic cell nuclear transfer embryos. One approach to produce autologous, disease-free ES cells from patients that may resolve these problems is the derivation of uniparental embryonic stem cells from the patient's gametes. Uniparental embryos such as parthenogenetic embryos have limited developmental capacity due to genomic imprinting but can produce ES cell lines. We have established that hematopoietic stem cells derived from murine uniparental ES cells can reconstitute adult hematopoiesis with no apparent pathology. To investigate the potential of uniparental cells for liver replacement, we now propose to use maternally (oocyte)-derived (parthenogenetic /gynogenetic) and paternally derived (androgenetic; two sperm genomes) fetal ES cell derivatives to functionally repopulate the liver in fumarylacetoacetate hydrolase (Fah) deficient adults. As a therapeutic approach, we will also transplant hepatic progenitors derived in vitro from uniparental ES cells. As each gamete contains a subset of the genome, diploid uniparental embryos are homozygous at a proportion of loci that are heterozygous in the gamete donor. This can be exploited to produce patient-derived ES cells without an errant allele (including large genetic lesions) in diseases associated with heterozygosity. Using the heterozygous phenotypic PiZ mouse model for alpha-1-antitrypsin deficiency, we will perform a proof of principle experiment involving elimination of the PiZ locus in uniparental ES cells. For patients of recessive genetic disorders, mutant allele-free, uniparental ES cell lines derived from parents or siblings could provide immune-compatible transplantable tissue, since MHC homozygous, mutant allele-free, uniparental lines with a matched subset of the recipient's MHC could be identified. Using mouse strains with different MHC loci, we will investigate the engraftment of MHC homozygous cellular liver transplants in MHC heterozygous recipients, i.e. determine the relevance of hybrid resistance.This proposal will investigate the capacity of embryonic stem cells derived from oocytes or sperm only to be used for liver transplantation and correction of genetic liver diseases. These embryonic stem cells would be derived from the respective patient's sperm or oocytes, and thus limit rejection problems associated with the use of existing embryonic stem cell lines.

描述（由申请人提供）： 对于许多肝脏疾病，包括先天遗传性肝脏疾病，供体器官移植是目前唯一的治疗选择。作为组织的替代来源，从胚胎干 (ES) 细胞衍生的细胞肝移植需要是自体的，并且没有遗传疾病，这些要求不容易克服，而且对于来自潜在可行的受精或患者特异性体细胞核移植胚胎的 ES 细胞来说，在伦理上也存在争议。从患者身上产生自体、无病 ES 细胞（可能解决这些问题）的一种方法是从患者的配子中衍生出单亲胚胎干细胞。单亲胚胎（例如孤雌胚胎）由于基因组印记而具有有限的发育能力，但可以产生 ES 细胞系。我们已经确定，源自小鼠单亲 ES 细胞的造血干细胞可以在没有明显病理的情况下重建成体造血功能。为了研究单亲细胞用于肝脏替代的潜力，我们现在建议使用母体（卵母细胞）衍生的（孤雌生殖/雌核遗传）和父本衍生的（雄激素遗传；两个精子基因组）胎儿ES细胞衍生物在富马酰乙酰乙酸水解酶（Fah）缺陷的成人中功能性地重新填充肝脏。作为一种治疗方法，我们还将移植来自单亲 ES 细胞的体外肝祖细胞。由于每个配子包含基因组的一个子集，二倍体单亲胚胎在配子供体中杂合的部分基因座上是纯合的。这可用于在与杂合性相关的疾病中产生患者来源的 ES 细胞，而没有错误的等位基因（包括大的遗传损伤）。使用 α-1-抗胰蛋白酶缺乏症的杂合表型 PiZ 小鼠模型，我们将进行原理验证实验，涉及消除单亲 ES 细胞中的 PiZ 基因座。对于隐性遗传病患者，源自父母或兄弟姐妹的无突变等位基因的单亲 ES 细胞系可以提供免疫相容的可移植组织，因为可以鉴定出具有与受体 MHC 匹配子集的 MHC 纯合、无突变等位基因的单亲细胞系。我们将使用具有不同MHC位点的小鼠品系，研究MHC纯合细胞肝移植在MHC杂合受体中的植入，即确定杂种抗性的相关性。该提案将研究源自卵母细胞或精子的胚胎干细胞仅用于肝移植和遗传性肝病的纠正的能力。这些胚胎干细胞将源自相应患者的精子或卵母细胞，从而限制与使用现有胚胎干细胞系相关的排斥问题。