Pro-Inflammatory Stem Cell Variants in Cystic Fibrosis

囊性纤维化中的促炎干细胞变异体

• 批准号: 10557166
• 负责人: FRANK D. MCKEON
• 金额: $ 67.81万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10557166
• 关键词: Antibiotics; Bacteria; Bacterial Infections; CRISPR/Cas technology; Cells; Chronic; Chronic Obstructive Pulmonary Disease; Clinical; Cloning; Complement; Complex; Coupled; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Dependence; Development; Disease; Disease Progression; Drug Modulation; Elements; Epithelial Cells; Excision; Ferrets; Fetal Lung; Fibrosis; Gene Expression; Genes; Genotype; Health; Human; Individual; Inflammation; Inflammatory; Kinetics; Link; Lung; Lung diseases; Methods; Modeling; Monitor; Mutation; Neutrophil Infiltration; Nodal; Organ; Pathogenicity; Pathology; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Play; Process; Proliferating; Property; Pulmonary Cystic Fibrosis; Pulmonary Inflammation; Role; Testing; Therapeutic; Time; VX-770; Variant; Withdrawal; Xenograft Model; Xenograft procedure; airway epithelium; cystic fibrosis infection; cystic fibrosis patients; gene complementation; genetic signature; genome editing; improved; in utero; inflammatory lung disease; insight; microbial colonization; mucin hypersecretion; mutant; neutrophil; novel; novel therapeutics; pathogenic bacteria; prospective; pulmonary function; respiratory smooth muscle; response; restoration; smoking abstinence; smoking cessation; stem cells; therapeutic target

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.

炎症在囊性纤维化（CF）的进行性病理学中起主要作用，通常被认为是 对CF肺的微生物定殖的增加反应。但是，最近的研究涉及正常和 尽管 缺乏细菌病原体。此外，虽然CFTR调节器的革命类别可以改善肺 功能并减少加重，它们在减轻CF肺的炎症方面的成功率较低。这些 发现提出了炎症，也许是CF的其他病原特征的可能性 疾病中出现的元素，但随后驱动与CFTR活性无关。类似的 情景可能在慢性阻塞性肺疾病（COPD）中进行，其中炎症和疾病 尽管戒烟，进展仍在继续。在COPD中，最近的研究表明相关性很强 （p <10-16）在促炎，小气道上皮细胞的出现与疾病本身之间。 这些致病性变异也存在，尽管在没有COPD的对照患者和胎儿的对照患者中，但也存在 肺。对CF肺的类似分析表明，它们被致病的干细胞变体淹没了 与在COPD中看到的那些以及两个以前未在 COPD肺。我们假设这些CF干细胞变体在CF的进展中起关键作用，并且 代表该疾病的致病元素，这些疾病是由CFTR基因型触发的，但却无关的。到 检验这一假设，并扩展我们对这些变体在CF疾病中潜在意义的理解 流程，我们将在三个特定的目标中，1）确定三种高炎性的关键炎症驱动因素 使用CRISPR-CAS9指导突变和异种移植模型的人类CF变体，2）测试 使用基因的CF患者在CFTR活性中发现的这三种变体的促炎表型 互补和CFTR调节药物，例如ivacaftor，Elexacaftor和Tezacaftor，3）利用我们的 最近开发了克隆雪貂气道干细胞来确定致病动力学的方法 在CF进展的条件模型中的变体。我们预计这些研究将提供背景 并深入了解占主导肺的变体干细胞的贡献，评估新的影响 CF治疗这些细胞的致病特征，并鉴定炎症中的淋巴结基因 这些变体的签名可能对这些患者具有治疗益处。