Studying a bone marrow failure disease using patient-specific iPS cells

使用患者特异性 iPS 细胞研究骨髓衰竭疾病

基本信息

批准号：
8819563
负责人：
Luis Francisco Zirnberger Batista
金额：
$ 24.28万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-03-10 至 2017-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8819563
关键词：
Adult Affect Age Aging Aplastic Anemia Binding Blood Cells Bone Marrow Cell Line Cell physiology Cells ChIP-seq Chromatin Chromosomes Clinical Clinical Protocols Complement Complex Custom DNA-Directed DNA Polymerase Defect Development Disease Drug Targeting Dyskeratosis Congenita Event Functional disorder Future Gene Expression Profiling Genes Genetic Goals Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells High-Throughput Nucleotide Sequencing Homeostasis Human Inherited Knowledge Length Life Life Expectancy Link Mammalian Cell Methods Modeling Molecular Molecular Profiling Mus Mutation Nail plate Organ Outcome Pancytopenia Pathway interactions Patients Pharmaceutical Preparations Phase Phenotype Physiological Population Preclinical Drug Evaluation Process Pulmonary Fibrosis RNA Sequences Regenerative Medicine Regulation Rest Ribonucleoproteins Severities Signal Transduction Skin Somatic Cell Stem cells System Technology Telomerase Telomere Maintenance Telomere Shortening Tissues adult stem cell alternative treatment biochemical tools bone marrow failure syndrome cell type cellular development coping design disease phenotype drug discovery high throughput analysis human tissue induced pluripotent stem cell mutant novel novel strategies patient population premature research study response screening self-renewal small molecule stem stem cell division stem cell technology telomere telomere loss therapy development transcriptome sequencing

项目摘要

7. PROJECT SUMMARY/ABSTRACT The goal of this project is to use patient-specific induced pluripotent stem cells (iPSCs) as a platform for the development of novel therapies for patients suffering with dyskeratosis congenita (DC), a bone-marrow failure syndrome that presents with poor life expectancy and multi-systemic tissue defects that include aplastic anemia and pulmonary fibrosis. A combination of technologies will be used to achieve this goal, including genetic correction, high-throughput sequencing, small-molecule drug screening and targeted differentiation of DC iPS cells to hematopoietic fates. All patients with DC have very short telomeres for their age, typically below the first percentile length when compared to the rest of the population. All mutations discovered in DC so far were in genes related to telomere homeostasis, either in telomerase, or directly binding to telomeres. Telomerase is the multi-enzymatic complex responsible for telomere synthesis in mammalian cells. In the absence of telomerase, telomeres will progressively shorten, which has been linked to impaired stem cell function in mice and humans. Thus, the tissue defects observed in DC likely result from the loss of self-renewal in adult stem cells compartments of these patients, caused by accelerated telomere shortening in settings of mutant telomerase. The central hypothesis of this project is that disease-specific iPS cells offer a novel and suitable system to study the consequences of mutant telomerase and the loss of self-renewal in DC pluripotent cells and can be used to search for strategies to correct this phenotype. I have previously shown that iPS cells derived from patients showing variable severity of DC faithfully recapitulate the telomere shortening and loss of self-renewal phenotypes observed in human patients. Here, I propose to use these patient-specific iPS cells to understand in detail the loss of self-renewal phenotype arising from dysfunctional telomeres and to use these cells as a platform for drug discovery and targeted differentiation, which could enable novel protocols for clinical therapy in the future. The Specific Aims are (I) to reverse the self-renewal defect in DC patient-specific iPS cells by genetic complementation and high- throughput gene expression analysis of DC iPS cells with critically short telomeres and (II) to use DC iPS cells as a platform for developing new therapies against DC, by searching for telomerase stabilizing drugs and by specifically differentiating these cells into hematopoietic fates. This project will significantly increase the current knowledge on bone-marrow failure syndromes, by first understanding the deleterious effects of dysfunctional telomeres in human pluripotent cells, and then by devising novel strategies to treat patients afflicted with dyskeratosis congenita, a disease that currently has no cure.

7。项目摘要/摘要该项目的目的是使用特定于患者的诱导多能干细胞（IPSC）作为平台为患有疾病的患者（DC）的患者开发新的疗法，这是一种骨髓失败综合征的预期寿命和多系统组织缺陷，包括性质贫血和肺纤维化。将使用多种技术来实现这一目标，包括遗传校正，高通量测序，小分子药物筛查和针对性的分化 DC IPS细胞致造血命运。所有DC患者的年龄都非常短，通常低于第一个百分位长度与其他人口相比。到目前为止，在DC中发现的所有突变都与与端粒稳态，无论是端粒酶，还是直接与端粒结合。端粒酶是多酶复合物负责哺乳动物细胞中端粒合成。在没有端粒酶的情况下，端粒将逐渐缩短，这与小鼠和人类的干细胞功能受损有关。因此， DC中观察到的组织缺陷可能是由于成年干细胞隔室中自我更新的丧失而导致的这些患者是由突变端粒酶环境中加速的端粒缩短引起的。中央该项目的假设是疾病特异性的IPS细胞提供了一种新颖而合适的系统来研究突变端粒酶的后果和直流多能细胞中自我更新的丧失，可用于寻找纠正此表型的策略。我以前已经表明，源自患者的IPS细胞忠实地表现出可变的DC严重程度概括人类患者观察到的端粒缩短和自我更新表型的丧失。在这里，我建议使用这些特定患者的IPS细胞详细了解自我更新表型的丧失由功能失调的端粒引起，并将这些细胞用作药物发现的平台并针对性分化，这可以使未来的临床治疗方案实现新颖的方案。具体目的是（i）通过遗传互补和高 - DC IPS细胞的吞吐量基因表达分析，其端粒和（ii）使用DC IPS细胞作为开发针对DC的新疗法的平台，通过寻找端粒酶稳定药物和通过专门将这些细胞区分为造血命运。首先了解人类多能细胞中功能失调的端粒的有害作用，然后通过制定新的策略来治疗患有疾病症患者的患者，这种疾病目前尚无治愈。