Novel Strategies to Clear Bacteria from the CF Lung

清除 CF 肺细菌的新策略

• 批准号: 10680454
• 负责人: Saira Ahmad
• 金额: $ 97.78万
• 依托单位: ELDEC PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-10 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10680454
• 关键词: Aerosols; Affect; Alveolar Macrophages; Antibiotic Resistance; Antimicrobial Effect; Bacteria; Bacterial Antibiotic Resistance; Bacterial Infections; Biological Models; Birth; Burkholderia cepacia; Burkholderia cepacia complex; Capital; Cell membrane; Cells; Chronic; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Cystic Fibrosis sputum; Data; Dehydration; Disease Progression; Dose; Drug Kinetics; Effector Cell; Equilibrium; Ferrets; Foundations; Genes; Genetic Diseases; Glosso-Sterandryl; Haemophilus influenzae; Immune; Immune response; Immunocompromised Host; Immunology; Immunomodulators; Inflammation; Inflammatory Response; Inhalation; Iowa; Leukocyte Elastase; Lung; Lung diseases; Lung infections; Mediating; Modeling; Mucous body substance; Mus; Mutation; Nasal Epithelium; Nebulizer; Neutrophilia; Nonsense Mutation; Normal Range; Palate; Patients; Peptide Hydrolases; Peptides; Peripheral; Play; Population; Privatization; Protein Secretion; Proteins; Pseudomonas; Pseudomonas aeruginosa; Public Health; Pulmonary Cystic Fibrosis; Pulmonary Function Test/Forced Expiratory Volume 1; Pulmonary Inflammation; Reagent; Regimen; Regulator Genes; Role; Sodium Chloride; Staphylococcus aureus; System; Testing; Therapeutic; Translating; Universities; Water; aerosolized; antimicrobial; cell type; chronic infection; clinical development; combat; cystic fibrosis airway; cystic fibrosis infection; cystic fibrosis mouse; cystic fibrosis patients; cytokine; disease-causing mutation; experimental study; improved; lung health; lung injury; methicillin resistant Staphylococcus aureus; multidrug-resistant Pseudomonas aeruginosa; neutrophil; novel; novel strategies; pathogen; peptidomimetics; prevent; programs; pulmonary function

Cystic fibrosis (CF) is a genetic disease caused by mutations in the cystic fibrosis transmembrane regulator (CFTR) gene. CF airways are immunocompromised and become colonized with bacteria soon after birth. Chronic bacterial infection leads to persistent and severe neutrophil-dominated pulmonary inflammation, high lung protease levels, lung damage and a decline in FEV1. CFTR modulator/correctors from Vertex such as TRIKAFTA significantly increase CF patient lung function by >10% but do not bring it into the normal range and for patients with pre-existing bacterial lung infections, do not clear bacteria from their lungs. Moreover, these compounds do not treat CF patients with nonsense mutations where no CFTR protein is produced. Thus, there is a critical unmet need for novel, CFTR mutation-agnostic therapies to help clear bacteria from CF lungs and limit neutrophilic lung damage. Short Palate LUng and Nasal epithelial Clone 1 (SPLUNC1) is a secreted protein that is highly expressed in the lung, where it plays a key role in maintaining lung health. SPLUNC1 is a CF gene modifier, and patients with reduced SPLUNC1 levels have lower FEV1 and exacerbate more frequently. Orai1 is a ubiquitously expressed plasma membrane Ca2+ channel that regulates inflammation. We found that SPLUNC1 inhibits Orai1. However, SPLUNC1 is rapidly degraded by neutrophil elastase (NE), which we posit results in greater Orai1 activity and more inflammation. Consistent with this, our preliminary data indicate that Orai1 is upregulated in CF patient lung immune cells. Given Orai1’s proximal role in the immune response, Orai1 is thus an attractive target whose inhibition is predicted to help resolve CF inflammation. Eldec Pharma has developed a robust, novel peptidomimetic called ELD607, which reprises SPLUNC1’s ability to inhibit Orai1, yet is significantly more stable in the presence of NE, and significantly more potent/efficacious. ELD607 is stable in proteolytic CF sputum, and inhibits Ca2+-influx in freshly-isolated CF patient peripheral neutrophils and in CF sputum-derived immune cells in a mutation-agnostic fashion. In murine lung infection models with common CF pathogens including P. aeruginosa and S. aureus, a single, inhaled dose of ELD607 reduced lung inflammation (neutrophilia, cytokines, NE) by 90%, decreased lung bacterial infection by 3-4 log10 CFUs and increased survival. In a chronic CF model (SCNN1B mice), ELD607 reduced neutrophilia and increased survival. These experiments demonstrate that rebalancing the lung’s inflammatory response by inhibiting Orai1 enhances the lungs’ natural ability to clear pathogens. In this proposal, we will use wild-type and CF mice to understand which immune effector cells are regulated by ELD607. The CF ferret model developed by Dr Engelhardt and coworkers at the University of Iowa spontaneously develops chronic CF lung disease including inflammation and bacterial infection. We will first validate that we can deliver sufficient doses of ELD607 via nebulizer to safely inhibit Orai1 in wild-type and CF ferret lungs. Then we will chronically administer ELD607 to CF ferrets with existing lung disease in order to study the impact of ELD607 on CF disease progression.

囊性纤维化（CF）是一种遗传性疾病，由囊性纤维化跨膜调节因子突变引起 （CFTR）基因。CF气道免疫功能低下，出生后不久就被细菌定植。 慢性细菌感染导致持续和严重的嗜中性粒细胞为主的肺部炎症，高 肺蛋白酶水平、肺损伤和FEV 1下降。来自Vertex的CFTR调制器/校正器，例如 TRIKAFTA显著增加CF患者肺功能>10%，但未将其带入正常范围， 对于已经存在细菌性肺部感染的患者，不要清除肺部的细菌。而且这些 化合物不治疗具有无义突变的CF患者，其中不产生CFTR蛋白。因此 是对新型CFTR突变不可知疗法的关键未满足需求，以帮助清除CF肺中的细菌， 限制嗜酸性肺损伤。短腭肺和鼻上皮克隆1（SPLUNC 1）是一种分泌蛋白， 它在肺部高度表达，在维持肺部健康方面起着关键作用。SPLUNC 1是CF基因 SPLUNC 1水平降低的患者FEV 1较低，且更频繁地恶化。Orai 1是 一种普遍表达的调节炎症的质膜钙通道。我们发现SPLUNC 1 抑制Orai 1。然而，SPLUNC 1被中性粒细胞弹性蛋白酶（NE）迅速降解，我们发现这导致了 Orai 1活性更高，炎症更多。与此一致，我们的初步数据表明Orai 1是 在CF患者肺免疫细胞中上调。鉴于Orai 1在免疫应答中的近端作用，因此Orai 1是 一个有吸引力的目标，其抑制预计有助于解决CF炎症。Eldec Pharma开发了 一种名为ELD 607的强大的新型肽模拟物，它重现了SPLUNC 1抑制Orai 1的能力， 在NE的存在下显著更稳定，并且显著更有效。ELD 607在以下环境中稳定 蛋白水解CF痰液，并抑制新鲜分离的CF患者外周中性粒细胞和CF中的Ca 2+内流 以突变不可知的方式研究肿瘤衍生的免疫细胞。在具有常见CF的小鼠肺部感染模型中 病原体包括铜绿假单胞菌和S.金黄色葡萄球菌，单次吸入剂量的ELD 607减少了肺部炎症 （嗜中性粒细胞，细胞因子，NE）减少90%，肺部细菌感染减少3-4 log 10 CFU， 生存在慢性CF模型（SCNN 1B小鼠）中，ELD 607减少了嗜中性粒细胞，并增加了存活率。这些 实验表明，通过抑制Orai 1来重新平衡肺部的炎症反应可以增强 肺部清除病原体的天然能力。在这个提议中，我们将使用野生型和CF小鼠来了解 免疫效应细胞受ELD 607调节。恩格尔哈特博士及其同事开发的CF雪貂模型 在爱荷华州大学， 感染我们将首先验证我们可以通过雾化器输送足够剂量的ELD 607以安全地抑制Orai 1 在野生型和CF雪貂肺中。然后，我们将长期给予ELD 607与现有的肺CF雪貂 为了研究ELD 607对CF疾病进展的影响，