Lung epithelial cell specification in human pluripotent stem cells

人多能干细胞的肺上皮细胞规格

• 批准号: 10378129
• 负责人: HANS-WILLEM E SNOECK
• 金额: $ 56.79万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378129
• 关键词: Address; Affect; Air; Airway Disease; Basal Cell; Benchmarking; Bleomycin; Cell Therapy; Cell model; Cells; Characteristics; Clonality; Clone Cells; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Disease; Distal; Engraftment; Epithelial; Epithelial Cells; Fetal Lung; Future; Generations; Genome Stability; Goals; Goblet Cells; Home; Human; Immunodeficient Mouse; In Vitro; Injury; Interphase; Length; Liquid substance; Lung; Lung Transplantation; Lung diseases; Measures; Modeling; Molecular; Mus; Naphthalene; Organ Donor; Pathogenesis; Patients; Pharmaceutical Preparations; Phenotype; Play; Population; Population Heterogeneity; Procedures; Replacement Therapy; Reporting; Role; Source; Telomerase; Time; Trees; alveolar epithelium; cell replacement therapy; curative treatments; embryonic stem cell; experimental study; fetal; genome-wide; human pluripotent stem cell; idiopathic pulmonary fibrosis; in vivo; indium-bleomycin; induced pluripotent stem cell; injured airway; irradiation; lung injury; progenitor; reconstitution; senescence; stem cells; surfactant deficiency; telomere; trafficking

Lung transplantation, the only definitive treatment for end-stage lung disease, remains hampered by severe complications and a shortage of donor organs. One potential alternative for at least some diseases where the epithelium is specifically affected is stem cell or cellular replacement therapy. Application of this type of therapy is still far away, however. Prerequisites for cellular therapy are an unlimited supply of the correct epithelial progenitors and an efficient engraftment procedure. In this proposal, we focus primarily on the first goal, and use generic injury models to demonstrate the potential of these cells. To address the issue of an unlimited supply of potential airway progenitors, we focused on embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), collectively termed human pluripotent stem cells (hPSCs). We describe here the generation from hPSCs of a population of putative fetal lung distal tip progenitors (pDTPs) that can be expanded continuously (up to now 18 months) and efficiently repopulates most or all lineages in bleomycin-injured lungs of immunodeficient NSG mice. Furthermore, these cells can be converted to cells with all the characteristics of basal cells (BCs), the stem cells of the airway, which, however, showed more limited expansion. Despite the fact the pDTPs engrafted in the airways of bleomycin-treated NSG mice, the derived BC-like cells did not engraft, suggesting that BCs may not possess engraftment capacity or that bleomycin is not an appropriate injury model for these cells. The goal of this proposal is to molecularly characterize both cell populations and evaluate their potentials in a variety of injury models. The specific aims of the proposal are: Aim 1: Molecular and in vitro characterization of pDTPs and BC-like cells Aim 2. In vivo potential of hPSC-derived pDTPs and BC-like cells. Aim 3. Culture conditions and clonality of pDTPs.

肺移植是终末期肺部疾病的唯一明确治疗 受到严重并发症和供体器官短缺的阻碍。一个潜力 至少有特异性影响上皮的某些疾病的替代性是茎 细胞或细胞替代疗法。这种类型的疗法的应用仍然很远， 然而。细胞疗法的先决条件是正确上皮的无限供应 祖细胞和有效的植入程序。在此提案中，我们主要关注 第一个目标，并使用通用伤害模型来证明这些细胞的潜力。 为了解决无限供应潜在气道祖细胞的问题，我们专注于 在胚胎干细胞（ESC）和诱导多能干细胞（IPSC）上 称为人类多能干细胞（HPSC）。我们在这里描述这一代人 假定的胎儿肺远端尖端祖细胞（PDTP）的HPSC可以是 不断扩展（直到现在长达18个月），并有效地重新占据大多数或全部 免疫缺陷型NSG小鼠的博来霉素造成的肺部谱系。此外，这些 可以将细胞转换为具有基底细胞（BC）的所有特征的细胞，茎 然而，气道的细胞显示出更有限的膨胀。尽管事实是 PDTP植入了博来霉素处理的NSG小鼠的气道中，该小鼠是衍生的BC样细胞 没有植入，表明BC可能没有植入能力或 博来霉素不是这些细胞的合适损伤模​​型。该提议的目的是 分子表征两个细胞群体，并评估其潜力 伤害模型。该提案的具体目的是： AIM 1：PDTP和BC样细胞的分子和体外表征 AIM 2。源自HPSC衍生的PDTP和BC样细胞的体内潜力。 目标3。培养条件和PDTP的克隆性。