Defining the genetic program of primordial lung progenitors

定义原始肺祖细胞的遗传程序

基本信息

批准号：
8975795
负责人：
Laertis Ikonomou
金额：
$ 42.85万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-21 至 2017-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8975795
关键词：
Adult Respiratory Distress Syndrome Affect Air Anterior Bronchopulmonary Dysplasia Cell Line Cell Separation Cell Therapy Cell Transplantation Cells Chronic Obstructive Airway Disease Complementary DNA Cues Cystic Fibrosis DNA Microarray Chip Data Derivation procedure Development Disease ES Cell Line Embryo Endoderm Epigenetic Process Epithelial Epithelial Cells Epithelium Ethical Issues Fibroblast Growth Factor Flow Cytometry Fluorescence Foundations Gene Expression Profile Genes Genetic Genetic Programming Goals Health Homeostasis Hour Human In Vitro Kidney Knock-in Knock-in Mouse Ligands Liquid substance Liver Lung Lung Transplantation Lung diseases Mesenchyme Messenger RNA Methods Modeling Morbidity - disease rate Mus Organ Outcome Pancreas Pathway interactions Patients Pattern Phenotype Pluripotent Stem Cells Population Primitive foregut structure Primordium Protocols documentation Publishing Pulmonary Emphysema Reagent Recombinants Replacement Therapy Reporter Respiratory Primordium Role Signal Pathway Signal Transduction Specific qualifier value Staging Stem cells Structure System Testing Thyroid Gland Time Tissues To specify Transplantation Tube base capsule design embryonic stem cell histone modification homologous recombination human embryonic stem cell in vivo induced pluripotent stem cell lung development methylation pattern mortality mouse development novel progenitor programs promoter reconstitution research study respiratory self-renewal stem stem cell differentiation thyroid transcription factor 1 tool transcription factor transcriptome

项目摘要

DESCRIPTION (provided by applicant): Diseases affecting the lung's epithelium are not easily treatable and result in significant morbidity and mortality worldwide. Specialized stem cells with the potential to self-renew or give rise to differentiated, functional progeny have been proposed as a critical component of tissue homeostasis for many organs, including the lung. Different potential approaches for the use of stem cells for lung disease treatment include enhancement of endogenous stem cell differentiation or in vitro directed differentiation of stem cells to lung lineages followed by cell transplantation. Both approaches require that the identity and pathways of differentiation of lung stem or progenitor cells be known and well characterized. Embryonic stem (ES) cells have emerged in the last 10-15 years as a promising platform for the development of cell-based therapies, since they can transit through several defined stages in vitro to recapitulate mammalian development. In addition, induced pluripotent stem (iPS) cells offer an attractive alternative to human ES cells. iPS cells are easy to derive, are not fraught with ethical issues and offer the possibility of patient-specific therapies. Nevertheless, the presumptive lung progenitor cells in development have not yet been identified, and this represents a major hurdle limiting the use of ES/iPS cells for lung disease therapies. The overall objective of this project is to define the genetic program of the multipotent epithelial progenitors of the lung primordium. This represents the first step towards our long-term goal of developing cell-based therapies for diseases affecting the lung epithelium. Since the transcription factor Titf1 is the earliest known marker of lung development, we created a new genetic tool, the Titf1-GFP knock-in mouse. This mouse will be used to characterize the progenitor cells of the lungs and the thyroid, both Titf1-positive and of endodermal origin. Using this tool, we will be able in Aim 1 to isolate cells from lung primordium based on GFP fluorescence, define their genetic program by means of DNA microarrays and study the epigenetics of genes important in lung development. We will proceed in Aim 2 to examine the role of FGF and Wnt signaling in lung specification in vitro using the Titf1-GFP ES and iPS cell lines. FGF and Wnt ligands are known to induce lung fate within definitive endoderm in vivo and we will assess whether they have a similar function in vitro. In Aim 3, we will test the functionality of the in vitro derived lung progenitors using novel and established systems, such as a bioartificial lung and air-liquid interface culture respectively. We envision that the outcome of our studies will be to define the optimal protocol for derivation of lung progenitor cells from ES/iPS cells to be used in novel lung disease therapies.

描述（由申请人提供）：影响肺上皮的疾病不容易治疗，并且在全球范围内导致大量发病率和死亡率。专门的干细胞具有自我更新或引起分化的功能性后代的潜力，作为许多器官（包括肺）的组织稳态的关键成分。使用干细胞进行肺部疾病治疗的不同潜在方法包括增强内源性干细胞分化或在体外定向干细胞对肺谱系的分化，然后进行细胞移植。两种方法都要求已知并且表征肺茎或祖细胞分化的身份和途径。在过去的10 - 15年中，胚胎干细胞（ES）细胞已成为开发基于细胞的疗法的有前途的平台，因为它们可以在体外通过几个定义的阶段过境，以概括哺乳动物的发育。此外，诱导的多能干（IPS）细胞为人ES细胞提供了有吸引力的替代品。 IPS细胞很容易得出，不充满道德问题，并提供特定于患者疗法的可能性。然而，尚未确定发育中的假定肺祖细胞，这代表了限制ES/IPS细胞用于肺部疾病疗法的主要障碍。该项目的总体目的是定义肺部多态上皮祖细胞的遗传程序。这是我们朝着我们长期目标开发基于细胞的疗法的疾病的第一步。由于转录因子TITF1是最早的肺发育标记物，因此我们创建了一种新的遗传工具TitF1-GFP敲入小鼠。该小鼠将用于表征肺和甲状腺的祖细胞，包括Titf1阳性和内胚层起源。使用此工具，我们将能够在AIM 1中根据GFP荧光分离肺原始细胞，通过DNA微阵列定义其遗传程序，并研究对肺发育中重要的基因表观遗传学。我们将在AIM 2中继续使用TITF1-GFP ES和IPS细胞系在体外检查FGF和WNT信号传导在体外的作用。已知FGF和Wnt配体在体内诱导肺命运，我们将评估它们在体外是否具有相似的功能。在AIM 3中，我们将分别使用新型和已建立的系统（例如生物人工肺和空气界面培养物）测试体外衍生的肺祖细胞的功能。我们设想我们的研究结果将是定义从ES/IPS细胞推导肺祖细胞的最佳方案，这些方案用于新型肺部疾病疗法。