Pro-Inflammatory Stem Cell Variants in Cystic Fibrosis

炎症在囊性纤维化(CF)的进行性病理中起主要作用，通常被认为是 是对CF肺微生物定植增加的反应。然而，最近的研究涉及正常和 在广谱抗生素下饲养的cftr突变雪貂在cfr肺中表现出强烈的炎症，尽管 没有细菌病原体。此外，革命性的CFTR调节器在改善肺功能的同时 在减轻肺纤维化的炎症方面，它们的疗效较差。这些 研究结果提出，炎症，也许是其他的CF的致病特征，是由 疾病中出现的因素，但随后推动进展，不依赖CFTR活性。一种类似的 情景可能发生在慢性阻塞性肺疾病(COPD)中，炎症和疾病 尽管戒烟，病情仍在继续。在慢性阻塞性肺病方面，最近的研究表明， (P&lt；10-16)促炎小气道上皮细胞的出现与疾病本身之间的关系。 这些致病变异体在没有COPD的对照组患者和胎儿中也存在，尽管水平较低。 阿龙。一项类似的对CF肺的分析显示，它们被致病干细胞变体高度淹没 与COPD中看到的那些相关，以及两个以前没有发现的新的、高炎症变体 慢性阻塞性肺病的肺。我们假设这些CF干细胞变体在CF的进展中起着关键作用，并且 代表这种疾病的致病因素，由CFTR型引发，但又独立于CFTR型。至 验证这一假设，并扩大我们对这些变异在CF病中的潜在意义的理解 过程中，我们将在三个具体目标中，1)确定三个关键的炎症驱动因素，即高度炎症 使用CRISPR-Cas9导向突变和异种移植模型的人CF变异体，2)测试对 应用基因技术研究CF患者中这三种变异体的致炎表型 互补和cftr调节药物，如iVacaftor、elexaftor和tezacaftor，以及3)利用我们的 最近开发的克隆雪貂呼吸道干细胞的方法来确定病原菌的动态 Cf进展的雪貂条件模型中的变体。我们预计这些研究将提供背景 以及洞察主导CF肺的变异干细胞的贡献，评估新的 Cf治疗学对这些细胞的致病特性以及炎性结节基因的识别 这些变异的特征，其抑制可能对这些患者的治疗有益。

Inflammation plays a major role in the progressive pathology of cystic fibrosis (CF), and is generally thought to be a response to increased microbial colonization of CF lungs. However, recent studies involving normal and CFTR-mutant ferrets raised under broad-spectrum antibiotics show robust inflammation in the CF lung despite the absence of bacterial pathogens. Moreover, while the revolutionary class of CFTR modulators improve lung function and reduce exacerbations, they are less successful in mitigating inflammation of the CF lung. These findings raise the possibility that inflammation, and perhaps other pathogenic features of CF, are maintained by elements that emerge in the disease but then drive progression independent of CFTR activity. An analogous scenario may be operating in chronic obstructive pulmonary disease (COPD), where inflammation and disease progression continues despite smoking cessation. In COPD, recent studies have shown a strong correlation (p<10-16) between the emergence of pro-inflammatory, small airway epithelial cells and the disease itself. These pathogenic variants are also present, albeit at low levels, in control patients without COPD and in fetal lung. A similar analysis of CF lungs has revealed them to be inundated by pathogenic stem cell variants highly related to those seen in COPD, along with two novel, hyperinflammatory variants not previously identified in COPD lungs. We hypothesize that these CF stem cell variants play key roles in the progression of CF, and represent pathogenic elements of this disease triggered by, and yet independent of, the CFTR genotype. To test this hypothesis and extend our understanding of the potential significance of these variants in CF disease processes, we will, in three specific aims, 1) identify key inflammatory drivers in the three, hyperinflammatory human CF variants using CRISPR-Cas9-directed mutations and xenograft models, 2) test the dependence of the pro-inflammatory phenotype of these three variants found in CF patients on CFTR activity using gene complementation and CFTR-modulating drugs such as ivacaftor, elexacaftor, and tezacaftor, and 3) exploit our recently developed methods for cloning ferret airway stem cells to determine the dynamics of the pathogenic variants in a ferret conditional model of CF progression. We anticipate that these studies will provide context and insight into the contributions of variant stem cells that dominate CF lungs, assess the impact of the new CF therapeutics on the pathogenic features of these cells, as well as identify nodal genes in the inflammatory signatures of these variants whose suppression could be of therapeutic benefit to these patients.