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: 8714996
• 负责人: Nathan A Zaidman
• 金额: $ 2.9万
• 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/8714996
• 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）：人体气道内衬高度特化的假复层上皮，作为抵御吸入性病原体的第一道防线。该组织包含祖细胞和分化的细胞类型，其促进响应损伤的粘膜纤毛清除和上皮修复。上皮损伤可能导致慢性炎症、重塑和明显受损的防御功能，这些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。本提案中描述的研究的长期目标是从人诱导多能干细胞（iPS）开发生理响应和临床适用的组织工程气道。该项目将利用最近发表的从人干细胞中获得肺上皮的技术，以产生假复层上皮，该假复层上皮可以在临床相关基质上生长，用作外科组织移植物。本申请中提供的初步数据显示，我们已经能够将人iPS细胞分化为表达p63+/CK 5+的气道基底细胞样祖细胞。我们建议采用额外的分化程序，以产生一个完全分化的气道上皮细胞能够正常的离子转运功能和自分泌信号分子的调节。为了实现这一目标，提出了以下目的：目的1）将人iPS细胞分化为气道上皮祖细胞，所述气道上皮祖细胞可用于在脱细胞气道基质上产生假复层上皮;目的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性质;目的3）鉴定由分化的上皮细胞表达的关键受体介导的信号传导途径，所述信号传导途径已知对于气道上皮的离子转运功能的正常调节是必需的。这些研究的结果将提供关于从人iPS细胞开发临床适用的组织工程气道移植物的重要见解。此外，这些目标的成功完成不仅会对肺组织工程产生影响，而且会对其他器官系统产生影响，其中上皮细胞分化和运输功能对于临床应用至关重要。