Pathogenesis of Airway Stem Cell Abnormalities in Obliterative Bronchiolitis

闭塞性细支气管炎气道干细胞异常的发病机制

基本信息

批准号：
9900856
负责人：
Kalpaj Rajnikant Parekh
金额：
$ 58.96万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9900856
关键词：
3-Dimensional Abnormal Cell Address Allografting Animal Model Autologous Basal Cell Biological Assay Bronchiolitis Caring Cause of Death Cell Lineage Cell Therapy Cell surface Cells Clinical Data Denervation Development Disease Progression Distal Enterobacteria phage P1 Cre recombinase Epithelial Epithelium Ferrets Fibrosis Foundations Genes Gland Goals Human Immune Targeting Impairment In Vitro Injections Injury Investigation Knock-out Knowledge Lead Left Lobar Lobe Location Lung Lung Transplantation Lung diseases Maintenance Mammals Modeling Morbidity - disease rate Mus Myoepithelial Myoepithelial cell Organoids Outcome Pathogenesis Pathology Pathway interactions Patients Phenotype Play Property Publishing Rattus Regulation Reporter Research Reserve Stem Cell Role Severities Signal Transduction Stem Cell Development Stem cell transplant Structure Surface Testing Therapeutic Time Trachea Transgenes Transgenic Organisms Transplant Recipients Transplantation WNT Signaling Pathway Xenograft procedure airway epithelium cartilaginous editorial effective therapy epithelial stem cell epithelium regeneration exhaustion human model implantation in vivo injury and repair lung allograft lung regeneration mortality novel novel therapeutics overexpression pre-clinical prevent public health relevance regenerative repaired self-renewal stem cell niche stem cell therapy stem cells transcription factor transplant model

项目摘要

Project Summary: End-stage lung diseases are a major cause of morbidity and mortality worldwide. Lung transplantation is an excellent treatment option for patients with this condition, yet 50% of recipients die within five years due to the development of obliterative bronchiolitis (OB) in the allograft. Epithelial stem cell depletion is suggested to contribute to the development of OB; however, there is little research being performed to test this hypothesis, primarily due to the lack of animal models for OB that develop allograft pathology resembling that seen in humans. We recently developed a novel orthotopic lung transplant model in the ferret that models human OB very well. Using this model, we have shown for the first time in ferret and human allografts that the number of clonogenic K5+p63+ basal stem cells (BSCs) progressively declines in proximal and distal airways of the allograft as the severity of OB increases. Additionally, our research is the first to demonstrate that the proximal airway submucosal gland (SMG), a facultative niche for BSCs in the surface airway epithelium (SAE), is an early target of immune destruction in human and ferret allograft airways. In mice, the SMG stem cell niche serves only the trachea; however, in larger mammals such as humans and ferrets, SMGs are present throughout the cartilaginous airways. Using lineage tracing, we have shown that the myoepithelial cells (MECs) of SMGs are precursors of multipotent K5+p63+ BSCs in the SAE. During the development of OB, the destruction of SMG stem cell niches in the allograft occurs simultaneously with phenotypic and functional changes to multipotent K5+p63+ BSCs in the SAE. We hypothesize that destruction of the denervated SMG in the allograft, and thus depletion of MECs, leads to a decline in multipotent K5+p63+ BSCs in the SAE, and to increases in committed multipotent (K5+p63+K14+), bipotent (K5+K14+, p63+K14+) and unipotent (K14+) basal cells, all of which have a reduced capacity for self-renewal. The objective of the proposed research is to determine the functional significance of the phenotypic changes in the MECS and the lineage-committed basal cells and how denervation of the SMG alters Wnt signaling (Lef-1/TCF1) required for both the maintenance of the SMG stem cell niche and lineage commitment of glandular MECs to SAE BSCs in the setting of injury. Additionally, a major preclinical objective of this proposal is to elucidate the ability of stem cells to engraft into a transplanted lung and repair injury, thus laying the foundation for the development of stem cell therapy to delay or prevent OB in lung allografts. We will achieve these objectives by addressing the following specific aims: 1) Determine how destruction of the SMG stem cell niche contributes to depletion of K5+p63+ BSCs. 2) Identify consequences of lung denervation on airway stem cells and their niches. 3) Determine the potential for airway stem cell transplantation in preventing or delaying OB. We expect that delineating the mechanisms of stem cell depletion in the airways of transplanted lungs will be a major step forward in understanding OB pathogenesis and will inform the use of stem cells as a therapeutic approach to prevent or delay OB.

项目摘要：末期肺部疾病是全球发病率和死亡率的主要原因。肺移植是对于患有这种情况的患者的出色治疗选择，但由于有50％的接受者在五年内死亡同种异体移植物中闭孔性支气管炎（OB）的发展。建议上皮干细胞耗竭有助于OB的发展；但是，几乎没有研究来检验这一假设，主要是由于缺乏OB的动物模型，该模型发展出类似于中的同种异体移植病理学人类。我们最近在雪貂中开发了一种新型的原位肺移植模型，该模型对人OB进行了建模很好。使用此模型，我们在雪貂和人类同种异体移植物中首次表明了克隆原性K5+ p63+基础干细胞（BSC）在同种异体移植的近端和远端气道中逐渐下降随着OB的严重程度的增加。此外，我们的研究是第一个证明近端气道的研究粘膜下腺（SMG）是表面气道上皮（SAE）中BSC的辅助利基市场，是一个早期目标人类和雪貂同种异体呼吸道的免疫破坏。在小鼠中，SMG干细胞小众仅服务于气管;但是，在人类和雪貂等较大的哺乳动物中，SMG存在于整个过程中软骨气道。使用谱系跟踪，我们表明SMG的肌上皮细胞（MEC）是 SAE中多能K5+ p63+ bsc的前体。在OB的发展过程中，SMG的破坏同种异体移植中的干细胞壁细胞壁细胞壁细胞与多能的表型和功能变化同时发生 SAE中的K5+ P63+ BSC。我们假设毁灭同种异体移植物中的被灭绝的SMG，从而 MEC的耗竭，导致SAE中的多元K5+ p63+ bsc的下降，并增加了所致敬的多能（K5+p63+K14+），两位数（K5+K14+，p63+k14+）和单位（K14+）基底细胞，所有这些都有一个减少自我更新能力。拟议研究的目的是确定功能 MECS和谱系结合的基底细胞的表型变化的重要性以及如何进行神经 SMG的wnt信号传导（LEF-1/TCF1）都需要维护SMG干细胞生态位和在伤害的情况下，腺体MEC对SAE BSC的谱系承诺。另外，主要的临床前该建议的目的是阐明干细胞植入移植肺并修复的能力受伤，从而为干细胞疗法的发展奠定了基础，以延迟或预防肺同种异体移植。我们将通过解决以下特定目的来实现这些目标：1）确定如何 SMG干细胞生态位的破坏会导致K5+ P63+ BSC的耗竭。 2）确定后果气道干细胞及其壁ni的肺部神经。 3）确定气道干细胞的潜力预防或延迟OB的移植。我们预计会描绘干细胞耗竭的机理在移植肺的气道中，将是理解OB发病机理的重要一步告知使用干细胞作为预防或延迟OB的治疗方法。