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）：人类气道由高度专业的假上皮衬里，该上皮是对吸入病原体的第一道防线。该组织既包含祖细胞类型，又有分化的细胞类型，可响应损伤，促进粘膜纤毛清除和上皮修复。上皮的损害可能导致慢性炎症，重塑和明显妥协的防御功能，这是许多大型气道疾病的标志，包括囊性纤维化（CF），慢性阻塞性肺部疾病（COPD），哮喘和癌症。尽管进行了数十年的研究，但对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从人类诱导的多能茎（IPS）细胞中开发出生理响应式和临床适用的组织工程气道。该项目将利用最近发表的有关人类干细胞肺上皮衍生的技术来产生假性化的上皮，可在临床相关的底物上生长，以用作手术组织移植物。本应用程序中提供的初步数据表明，我们能够将人IPS细胞区分为p63+/ck5+表达气道基底细胞样祖细胞。我们建议采用其他分化程序来产生能够通过自分泌信号分子进行正常离子传输功能和调节的完全分化的气道上皮。为了实现这一目标，提出了以下目的：目标1）将人IPS细胞区分为气道上皮祖细胞，这些细胞可用于在脱皮的气道底物上产生伪分层的上皮；目标2）表征参与粘膜清除率的关键离子转运蛋白的表达和亚细胞定位，并评估分化的伪分层上皮细胞的跨皮转运特性； AIM 3）鉴定由分化的上皮细胞表达的关键受体介导的信号通路，这些信号通路对于正常调节气道上皮的离子传输功能至关重要。这些研究的结果将提供有关人IPS细胞临床适用的组织呼吸道移植的重要见解。此外，成功完成目标不仅会影响肺组织工程，而且对上皮细胞分化和运输功能对于临床应用至关重要的其他器官系统。