PROJECT 1: Germ Cell Differentiation from Human iPSCs and hEScs

项目 1：人类 iPSC 和 hESc 的生殖细胞分化

基本信息

批准号：
8839146
负责人：
Renee A Reijo Pera
金额：
$ 34.61万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8839146
关键词：
AZF1 gene Address Adult Age Animal Model Biological Models Cell Differentiation process Cell Line Cells Cellular biology Characteristics Chromosome Deletion Chromosome Segregation Chromosomes, Human, Y Clinic Clinical Clinical Research Complex Couples Data Defect Deletion Mutation Derivation procedure Development Diagnostic Dissection Drosophila genus Drosophila melanogaster Experimental Genetics Failure Family member Female Frequencies Gene Dosage Gene Expression Gene Family Genes Genetic Genetic Epistasis Genetic study Genome Germ Cells Germ Lines Haploidy Human Human Development Human Genetics Human Genome Human Identifications In Vitro Infertility Intervention Intracytoplasmic Sperm Injections Investments Lesion Link Maintenance Maps Meiosis Methods Molecular Genetics Morphogenesis Mus Mutagenesis Mutation Oocytes Partner in relationship Pathology Pathway interactions Phenotype Platelet Factor 4 Pluripotent Stem Cells Production Property Proteins Protocols documentation Publishing Quality of life RNA-Binding Proteins Regenerative Medicine Relative (related person)Research Role Scientific Advances and Accomplishments Sertoli cell only syndrome Somatic Cell Spermatogenesis Staging System Testing Therapeutic Translations Transplantation Turner&apos s Syndrome Universities Variant Woman Y Chromosome Yeasts base clinical application data modeling deletion analysis design experimental analysis fly gain of function gene function gene interaction genetic analysis genetic variant human embryonic stem cell human embryonic stem cell line human female human genome sequencing human male imprint in vivo induced pluripotent stem cell loss of function mutation male men methylation pattern novel novel diagnostics overexpression pluripotency sex sperm cell stem cell biology success tool transcription factor

项目摘要

A. Significance A1. Infertility. Historically, the quality of life of infertile couples has been greatly diminished by the loss of opportunity to conceive. However, in recent years, novel clinical interventions such as intracytoplasmic sperm injection have dramatically changed the outlook for some couples, particularly those with severe forms of infertility[23]. In parallel with clinical successes, there have also been ground-breaking scientific advances including sequencing of the human genome, derivation of human embryonic stem cell (hESC) lines, and reprogramming of adult human somatic cells to pluripotency[24-28]. Together, these advances now allow us to overcome two historically-insurmountable limitations in studies of human development: the Inaccessibility of early human development to exploration and the genetic-intractability of the genome during development. A2. Need to study human germ cell development. 10-15% of couples are infertile, yet little is known of underlying pathologies in men and women with poor germ cell production. Here, we propose to extend our previous studies in order to permit genetic analysis of human germ cell development and thus, enable novel basic and clinical studies and applications. There are several unique aspects to human germ cell development that merit this investment. First, genes and dosages required for human germ cell development differ from those of mice, including both autosomal and sex chromosomal genes and dosages[6,29-34]. Second, humans are rare among species in that infertility is common, with half of all cases linked to faulty germ cell development[35]. Moreover, pathologies associated with meiotic errors are numerous in humans relative to other species, with errors in meiotic chromosome segregation occurring in as many as 5-30% of human germ cells depending on sex and age[36]. This is in contrast to frequencies of 1/10000 cells in yeast, 1/1000 cells in flies, and 1/100 cells in mice. With recent advances, we now have the ability to incorporate new strategies in order to examine the specifics of human germ cell development. This will allow us to derive full benefit from the wealth of data from model systems such as the fly and the mouse, to begin to understand the complex genetics of human germ cell formation and differentiation. In seeking to understand germ cell biology, we also acknowledge that the ability to contribute to the germ line is a fundamental property that distinguishes pluripotent stem cells. For example, Han and colleagues recently demonstrated that by addition of a 5th factor to the commonly-used 4 factor mixture for reprogramming, ability to contribute to the germ line was significantly increased[37]. This was in spite of the fact that IPSCs derived from 5 factor-reprogramming were indistinguishable from standard iPSCs or mESCs in gene expression and markers of pluripotency. Thus, the research proposed here allows us to address fundamental questions regarding our germ line origins, function, and pathology and lays the groundwork for designing rational therapeutics and diagnostics to inform clinical decisions based on data obtained from model organisms, human genetic studies and direct experimental analysis of human germ cells. It also contributes to the related field of pluripotent stem cell biology and regenerative medicine.

A.意义 A1。不育。从历史上看，由于失去了不育夫妇的生活质量。受孕的机会。但是，近年来，新型的临床干预措施，例如胞质内精子注射已大大改变了一些夫妻的前景，尤其是那些严重形式的夫妻不育[23]。与临床成功同时，也有开创性的科学进步包括对人基因组的测序，人类胚胎干细胞（HESC）系的衍生物，以及将成年人类体细胞重新编程为多能性[24-28]。这些进步现在使我们能够克服人类发展研究中的两个历史界限限制：早期的人类发展探索和发展过程中基因组的遗传缩写性。 A2。需要研究人类生殖细胞的发育。 10-15％的夫妻是不育的，但对生殖细胞生产不良的男性和女性的基本病理。在这里，我们建议扩展我们的先前的研究是为了允许对人类生殖细胞发育的遗传分析，因此可以实现新颖基础和临床研究和应用。人类生殖细胞发育有几个独特的方面值得这项投资。首先，人类生殖细胞发育所需的基因和剂量与小鼠，包括常染色体和性染色体基因和剂量[6,29-34]。其次，人类是在物种之间很少见，因为不孕症很常见，其中一半与生殖细胞发育有故障有关[35]。而且，与其他物种相对于其他物种，与减数分裂错误相关的病理有很多减数分裂染色体隔离的错误在多达5-30％的人类生殖细胞中发生性别和年龄[36]。这与酵母中1/10000细胞的频率，1/1000个细胞和1/100个细胞相反在老鼠中。随着最近的进步，我们现在有能力合并新策略，以检查人类生殖细胞发育的细节。这将使我们能够从来自诸如苍蝇和小鼠之类的模型系统开始理解人类细菌的复杂遗传学细胞形成和分化。在寻求了解生殖细胞生物学时，我们也承认有助于种系的能力是区分多能干细胞的基本特性。为了例如，Han及其同事最近证明，通过将第五因子添加到常用4 因子混合物重编程，有助于生殖系的能力显着增加[37]。这是在源自5个因子重新编程的IPSC与标准IPSC没有区别的事实或MESC在基因表达和多能性的标记中。因此，这里提出的研究使我们能够解决有关我们的细菌线，功能和病理学的基本问题，并提出设计理性疗法和诊断的基础工作，以根据数据为临床决策提供信息从模型生物，人类遗传研究和人类生殖细胞的直接实验分析获得。它还有助于多能干细胞生物学和再生医学的相关领域。