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呼吸道免疫功能受损，出生后不久就被细菌侵占。 慢性细菌感染导致持续和严重的以中性粒细胞为主的肺部炎症， 肺蛋白酶水平、肺损伤和FEV1下降。来自顶点的CFTR调制器/校正器，例如 TRIKAFTA可使慢性阻塞性肺疾病患者的肺功能显著提高10%，但没有使其恢复到正常范围 对于先前存在细菌性肺部感染的患者，不要清除肺部的细菌。此外，这些 化合物不治疗无义突变的CF患者，因为这些突变不会产生CFTR蛋白。因此，在那里 是尚未得到满足的对新型CFTR突变不可知疗法的需求，以帮助清除CF肺和肺组织中的细菌 限制中性粒细胞肺损伤。短腭肺鼻上皮克隆1是一种分泌型蛋白 它在肺中高度表达，在维持肺健康方面发挥着关键作用。SPLunc1是一个Cf基因 SPLunc1水平降低的患者FEV1更低，病情恶化更频繁。ORAI1是 一种普遍表达的质膜钙通道，调节炎症。我们发现SPLunc1 抑制Orai1。然而，SPLunc1被中性粒细胞弹性蛋白酶(NE)迅速降解，我们假设结果是 Orai1活性更强，炎症更多。与此一致，我们的初步数据表明Orai1是 CF患者肺免疫细胞表达上调。鉴于Orai1的S在免疫反应中的近端作用，Orai1因此 一个有吸引力的靶点，其抑制被预测有助于缓解CF炎症。Eldec Pharma已经开发出 一种名为ELD607的强健的、新颖的模拟肽药物，它使SPLUN1的S具有抑制Orai1的能力，但它 在NE存在的情况下显著更稳定，并且显著更有效/有效。ELD607在 蛋白水解性CF痰，并抑制新鲜分离的CF患者外周中性粒细胞和CF中的钙内流 以一种突变不可知的方式检测来自痰的免疫细胞。普通CF型小鼠肺部感染模型的建立 包括铜绿假单胞菌和金黄色葡萄球菌在内的病原体，单次吸入ELD607可减少肺部炎症 (中性粒细胞、细胞因子、NE)减少90%，减少肺部细菌感染3-4log10CFU，并增加 生死存亡。在慢性慢性纤维化模型(SCNN1B小鼠)中，ELD607降低了中性粒细胞并提高了存活率。这些 实验表明，通过抑制Orai1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。在这项提案中，我们将使用野生型和CF小鼠来了解哪些 免疫效应细胞受ELD607调控。由Engelhardt博士和他的同事建立的CF雪貂模型 在爱荷华大学自发性地患上包括炎症和细菌在内的慢性慢性肺病 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，以安全地抑制Orai1 在野生型和CF雪貂的肺中。然后，我们将长期给有肺的CF雪貂使用ELD607 为了研究ELD607基因对CF疾病进展的影响。