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）是由囊性纤维化跨膜调节剂突变引起的遗传疾病 （CFTR）基因。 CF气道被免疫功能低下，并在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中性粒细胞为主的肺部感染，高 肺部保护酶水平，肺损伤和FEV1下降。来自顶点的CFTR调制器/校正器，例如 Trikafta将CF患者肺功能显着提高> 10％，但不会将其带入正常范围，并且 对于先前存在的细菌肺部感染的患者，请勿从其肺中清除细菌。而且，这些 化合物不治疗未产生CFTR蛋白的无义突变患者。那，那里 是对新型CFTR突变敏锐疗法的关键需求，以帮助清除CF肺的细菌和 限制嗜中性肺损伤。短口肺和鼻上皮克隆1（Splunc1）是一种分泌的蛋白质 这在肺中高度表达，它在维持肺部健康方面起着关键作用。 Splunc1是CF基因 修饰符，Splunc1水平降低的患者的FEV1较低，并且更频繁地加重。 Orai1是 一个普遍表达的调节注射注射的质膜Ca2+通道。我们发现Splunc1 抑制Orai1。然而，Splunc1被嗜中性粒细胞弹性酶（NE）迅速降解，我们会呈阳性 更大的Orai1活性和更多的炎症。与此一致，我们的初步数据表明Orai1是 在CF患者肺免疫球中上调。鉴于Orai1在免疫响应中的近端作用，因此是ORAI1 一个有吸引力的目标，其抑制作用被预测有助于解决CF注射。 Eldec Pharma已开发 一种强大的，新颖的肽含量，称为ELD607，它可以重新抑制Orai1的能力，但IS是 在NE存在的情况下，明显更稳定，并且具有明显的潜力/有效性。 ELD607稳定 蛋白水解CF痰，并抑制新鲜分离的CF患者外周嗜中性粒细胞和CF中的Ca2+influx 痰液衍生的免疫球以突变敏捷的方式。在带有常见CF的鼠肺感染模型中 病原体，包括铜绿假单胞菌和金黄色葡萄球菌，单个内部剂量的ELD607减少了肺注射 （嗜中性粒细胞，细胞因子，NE）降低90％，减少3-4 log10 cfus的肺细菌感染并增加 生存。在慢性CF模型（SCNN1B小鼠）中，ELD607降低了中性粒细胞和生存率。这些 实验表明，通过抑制ORAI1来重新平衡肺部炎症反应会增强 肺部清除病原体的自然能力。在此建议中，我们将使用野生型和CF小鼠了解哪个 免疫效应细胞由ELD607调节。由恩格哈特博士和同事开发的CF雪貂模型 爱荷华大学自发发展慢性CF肺部疾病，包括感染和细菌 感染。我们将首先验证我们可以通过雾化器提供足够剂量的ELD607以安全抑制Orai1 在野生型和CF雪貂肺中。然后，我们将长期对ELD607进行管理 疾病以研究ELD607对CF疾病进展的影响。