Engineering human iPS cells into an airway epithelium capable of ion transport

将人类 iPS 细胞改造为能够进行离子运输的气道上皮

• 批准号: 8885495
• 负责人: Nathan A Zaidman
• 金额: $ 2.94万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8885495
• 关键词: Address; Air; American; Asthma; Autocrine Communication; Basal Cell; Breathing; CSNK1A1 gene; Carrier Proteins; Cause of Death; Cell Differentiation process; Cell model; Cell surface; Characteristics; Chronic; Chronic Obstructive Airway Disease; Clinical; Complex; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cytokeratin-14 Staining Method; Data; Derivation procedure; Development; Disease; Endoderm; Engineering; Epithelial; Epithelial Cells; Epithelium; Excision; Exhibits; Goals; Human; Human Engineering; Immunofluorescence Immunologic; Inflammation; Injury; Ion Transport; Lead; Liquid substance; Lung; Maintenance; Malignant Neoplasms; Membrane Proteins; Messenger RNA; Movement; Mucociliary Clearance; Operative Surgical Procedures; Organ; Pathway interactions; Patients; Pattern; Physiological; Play; Procedures; Production; Property; Pseudostratified Epithelium; Publishing; Purinergic P1 Receptors; Receptor Activation; Receptor Signaling; Regulation; Relative (related person); Reporting; Research; Role; Signal Pathway; Signaling Molecule; Sodium Chloride; Solutions; Stem cells; Stenosis; Techniques; Therapeutic; Thymus Gland; Thyroid Gland; Tissue Engineering; Tissue Grafts; Tissues; Trachea; Transcript; Transplantation; Transplanted tissue; airway epithelium; apical membrane; aquaporin 3; basolateral membrane; body system; cell type; clinical application; clinically relevant; epithelial Na+ channel; human stem cells; induced pluripotent stem cell; insight; monolayer; pathogen; progenitor; public health relevance; receptor-mediated signaling; repaired; research study; response; response to injury; solute; stem; success; trait

DESCRIPTION (provided by applicant): The human airways are lined by a highly specialized pseudostratified epithelium, which acts as the first line of defense against inhaled pathogens. This tissue contains both progenitor and differentiated cell types that facilitate mucociliary clearance and epithelial repair in response to injury. Damage to the epithelium may lead to chronic inflammation, remodeling and markedly compromised defense function that are hallmarks of many large airway diseases including cystic fibrosis (CF), chronic obstructive pulmonary disease (COPD), asthma and cancer. Despite decades of research, an adequate clinical solution to these airway diseases remains elusive. The long term goal of the research described in this proposal is to develop a physiologically responsive and clinically applicable tissue-engineered airway from human induced pluripotent stem (iPS) cells. This project will utilize recently published techniques on derivation of pulmonary epithelium from human stem cells to produce a pseudostratified epithelium that can be grown on clinically relevant substrates for use as a surgical tissue graft. Preliminary data presented in this application shows that we have been able to differentiate human iPS cells into p63+/CK5+ expressing airway basal cell-like progenitor cells. We propose to employ additional differentiation procedures to produce a fully differentiated airway epithelium capable of normal ion transport function and regulation by autocrine signaling molecules. To accomplish this goal, the following aims are proposed: Aim 1) Differentiate human iPS cells into airway epithelial progenitor cells that can be used for generating a pseudostratified epithelium on decellularized airway substrates; Aim 2) Characterize the expression and subcellular localization of key ion transport proteins involved in mucociliary clearance and assess the transepithelial transport properties of the differentiated pseudostratified epithelium; Aim 3) Identify key receptor-mediated signaling pathways expressed by differentiated epithelial cells that are known to be essential for normal regulation of ion transport function of the airway epithelium. The results of these studies will provide important insight regarding the development of a clinically applicable tissue- engineered airway graft from human iPS cells. Moreover, successful completion of the aims will have an impact not only on pulmonary tissue-engineering but also on other organ systems where epithelial cell differentiation and transport function are essential for clinical applications.

描述（由申请人提供）：人类呼吸道内衬有高度特化的假复层上皮，它是抵御吸入病原体的第一道防线。该组织含有祖细胞和分化细胞类型，可促进粘液纤毛清除和上皮修复以应对损伤。上皮损伤可能导致慢性炎症、重塑和防御功能明显受损，这是许多大气道疾病的标志，包括囊性纤维化（CF）、慢性阻塞性肺病（COPD）、哮喘和癌症。尽管经过了数十年的研究，这些气道疾病的适当临床解决方案仍然难以捉摸。该提案中描述的研究的长期目标是利用人类诱导多能干（iPS）细胞开发一种具有生理反应性和临床适用性的组织工程气道。该项目将利用最近发表的从人类干细胞衍生肺上皮的技术来产生假复层上皮，该上皮可以在临床相关的基质上生长，用作手术组织移植物。本申请中提供的初步数据表明，我们已经能够将人 iPS 细胞分化为表达 p63+/CK5+ 的气道基底细胞样祖细胞。我们建议采用额外的分化程序来产生完全分化的气道上皮，该上皮能够具有正常的离子转运功能并通过自分泌信号分子进行调节。为了实现这一目标，提出以下目标： 目的 1) 将人 iPS 细胞分化为气道上皮祖细胞，可用于在脱细胞气道基质上生成假复层上皮；目标 2) 表征参与粘膜纤毛清除的关键离子转运蛋白的表达和亚细胞定位，并评估分化的假复层上皮的跨上皮转运特性；目标 3) 识别由分化上皮细胞表达的关键受体介导的信号通路，已知这些信号通路对于气道上皮离子转运功能的正常调节至关重要。这些研究的结果将为利用人类 iPS 细胞开发临床适用的组织工程气道移植物提供重要见解。此外，成功完成这些目标不仅会对肺组织工程产生影响，还会对其他器官系统产生影响，在这些器官系统中上皮细胞分化和转运功能对于临床应用至关重要。