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

• 批准号: 8353117
• 负责人: Luis Francisco Zirnberger Batista
• 金额: $ 12.4万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-15 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8353117
• 关键词: Adult; Affect; Age; Aging; Aplastic Anemia; Binding; Biochemical; 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 Delivery Systems; Dyskeratosis Congenita; Event; Functional disorder; Future; Gene Expression; Genes; Genetic; Goals; Hematopoietic; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; 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; Screening procedure; Severities; Signal Transduction; Skin; Somatic Cell; Stem cells; Syndrome; System; Technology; Telomerase; Telomere Maintenance; Telomere Shortening; Tissues; adult stem cell; alternative treatment; 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; self-renewal; small molecule; stem; stem cell division; stem cell technology; telomere; therapy development; tool

DESCRIPTION (provided by applicant): 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 Specifi 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.

描述（由申请人提供）：该项目的目的是使用患者特异性诱导的多能干细胞（IPSC）作为用于开发患有患有病毒障碍性偶像性偶像的患者（DC）的新疗法的平台，这是一种骨髓衰竭综合征，具有较差的预期型和多型系统性组织的缺陷，其中包括较差的型和Pullonical Anemia和Pullostry Anemia。技术的组合将用于实现此目标，包括遗传校正，高通量测序，小分子药物筛查以及DC IPS细胞对造血命运的靶向分化。 与其他人口相比，所有DC患者的年龄端粒的年龄非常短，通常低于第一个百分位长度。到目前为止，在DC中发现的所有突变都是与端粒稳态有关的基因，无论是端粒酶，还是直接与端粒结合。端粒酶是负责哺乳动物细胞中端粒合成的多酶复合体。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，在DC中观察到的组织缺陷可能是由于这些患者的成年干细胞隔室的自我更新丧失所致，这是由于突变端粒酶环境中加速的端粒缩短引起的。该项目的中心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酶的后果以及DC多能细胞中自我更新的丧失，并且可以用于搜索以纠正该表型的策略。 我先前曾证明，从患者中得出的IPS细胞表现出DC的严重程度可变，忠实地概括了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，我建议使用这些特异性IPS细胞详细了解端粒功能失调的自我更新表型的丧失，并将这些细胞用作药物发现的平台和靶向分化的平台，这可以在将来实现新的临床治疗方案。该特定目的是（i）通过遗传互补和高吞吐量的基因表达分析，对DC IPS细胞的高吞吐量分析扭转DC患者特异性IPS细胞中的自我更新缺陷，并通过搜索针对DC的平台来搜索特定的telemerase稳定性，并使用DC IPS细胞进行较短的端粒和（ii）使用DC IPS细胞作为一种平台，并将这些疗法用于稳定性稳定性。 该项目将通过首先了解人类多能细胞中功能障碍端粒的有害影响，然后通过制定新的策略来治疗患有患病障碍性疾病的患者的新型策略，这将大大提高当前对骨髓衰竭综合征的知识，这是一种目前无法治愈的疾病。