Optimal Derivation of Murine Embryonic Distal Airway Stem Cells

小鼠胚胎远端气道干细胞的优化衍生

• 批准号: 8289647
• 负责人: CHRISTINE M FINCK
• 金额: $ 34.38万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8289647
• 关键词: 3-Dimensional; Accounting; Air Sacs; Alveolar; Applications Grants; Biomimetics; Birth; Blood Vessels; Blood capillaries; Borderline Personality Disorder; Bronchopulmonary Dysplasia; Cell Differentiation process; Cells; Childhood; Coculture Techniques; Cues; Derivation procedure; Development; Discipline of obstetrics; Distal; Embryo; Endoderm; Endoderm Cell; Endothelial Cells; Environment; Epithelial; Epithelial Cells; Event; FGF2 gene; Fibroblast Growth Factor 2; Gases; Goals; Grant; Growth Factor; Hydrogels; In Vitro; Laboratories; Low Birth Weight Infant; Lung; Lung diseases; Mesenchymal; Methods; Mus; Neonatal; Nodal; Pathologic Processes; Pathway interactions; Perinatal mortality demographics; Premature Birth; Process; Production; Proteins; Pulmonary Mass; Pulmonary Pathology; Research Personnel; Serum; Stem cells; Structure of parenchyma of lung; Supplementation; Surgeon; Time; Tissue Engineering; Training; Transplantation; Undifferentiated; United States; activin A; alveolar epithelium; capillary; embryonic stem cell; fetal; improved; in vivo; innovation; novel; premature lungs; public health relevance; scaffold; small molecule; stem cell therapy; surfactant; three dimensional structure; transcription factor

DESCRIPTION (provided by applicant): The pathology of Pulmonary Hypoplasia (PH) includes reduced lung mass, insufficient ability of the lungs to stay expanded due to a deficiency in surfactant protein production, a poorly differentiated lining of the air sac (alveolar epithelium), and a reduction of gas exchange. Our long term goal is to alleviate neonatal PH using stem cell therapy to augment premature lung tissue with in vitro stem cell derived distal airway cells. The specific hypothesis is that recapitulation of key developmental events by providing transcription and growth factors will optimize derivation of distal airway cells. This includes a 2 step process with derivation of definitive endoderm followed by differentiation into distal airway cells. Furthermore, providing a 3-dimensional scaffold will enhance our yield of distal airway cells by providing important spatial cues. In the lung, distal airway development and integrity is essential for proper gas exchange, surfactant production and survival. The focus of this grant proposal is to explore methods of deriving distal airway cells from embryonic stem cells. Our specific aims include: 1: Optimize derivation of definitive endoderm cells from murine embryonic stem cells, specifically by manipulating the Wnt and nodal pathway or by providing small molecules. 2: Optimize derivation of distal airway cells through growth factor supplementation, specifically FGF2, 7,10. 3: Optimize spatial cues utilizing 3-dimensional hydrogel scaffolds with potential transplantation in vivo, time permitting. PUBLIC HEALTH RELEVANCE: Preterm delivery with resultant immature lungs (pulmonary hypoplasia) is a major problem in obstetrics and accounts for more than 70% of perinatal mortality. In 2004, 12.5% of births in the United States were preterm. These immature lungs have reduced numbers of mature epithelial cells which results in insufficient surfactant production (a protein used to keep the distal airways open) as well as decreased vasculature. We propose to utilize murine embryonic stem cells to derive distal airway cells for possible utilization in pulmonary hypoplasia. This exploratory study may help identify optimal methods of deriving distal airway cells that can potentially be utilized for treatment of pulmonary diseases such as pulmonary hypoplasia.

描述（由申请人提供）：肺发育不全（PH）的病理包括肺质量减少、由于表面活性蛋白产生不足而使肺保持扩张的能力不足、气囊内壁（肺泡上皮）分化差以及气体交换减少。我们的长期目标是通过干细胞疗法通过体外干细胞衍生的远端气道细胞增强早产肺组织来缓解新生儿肺PH。具体的假设是，通过提供转录和生长因子来重现关键发育事件将优化远端气道细胞的衍生。这包括一个两步过程，即定形内胚层衍生，然后分化为远端气道细胞。此外，提供 3 维支架将通过提供重要的空间线索来提高远端气道细胞的产量。在肺部，远端气道的发育和完整性对​​于适当的气体交换、表面活性剂的产生和生存至关重要。该拨款提案的重点是探索从胚胎干细胞衍生远端气道细胞的方法。我们的具体目标包括： 1：优化从鼠胚胎干细胞衍生定形内胚层细胞，特别是通过操纵 Wnt 和 nodal 途径或通过提供小分子。图 2：通过补充生长因子（特别是 FGF2, 7,10）优化远端气道细胞的衍生。图 3：在时间允许的情况下，利用具有体内移植潜力的 3 维水凝胶支架优化空间线索。 公共卫生相关性：早产导致肺部不成熟（肺发育不全）是产科的一个主要问题，占围产期死亡率的 70% 以上。 2004年，美国12.5%的新生儿是早产儿。这些不成熟的肺部成熟上皮细胞的数量减少，导致表面活性剂（一种用于保持远端气道开放的蛋白质）产生不足以及脉管系统减少。我们建议利用小鼠胚胎干细胞来衍生远端气道细胞，以用于肺发育不全的治疗。这项探索性研究可能有助于确定衍生远端气道细胞的最佳方法，这些细胞可用于治疗肺发育不全等肺部疾病。