Targeting Hyperplastic Airway Cells Using a Small Molecule Inhibitor

使用小分子抑制剂靶向增生性气道细胞

• 批准号: 9035020
• 负责人: Hitendra Singh Chand
• 金额: $ 32.7万
• 依托单位: LOVELACE BIOMEDICAL & ENVIRONMENTAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2016-08-12
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9035020
• 关键词: Address; Adverse event; Affect; Allergic; American; Animal Model; Apoptosis; Apoptotic; Asthma; Autoimmune Diseases; Autoimmune Process; BCL2 gene; BH3 Domain; Bioavailable; Biological; Breathing; Cell Death; Cell Proliferation; Cell Survival; Cells; Cessation of life; Characteristics; Chronic; Clinical Trials; Data; Development; Disease; Disease Management; Epithelial; Epithelial Cells; Epithelium; Equilibrium; Event; Excision; Extrinsic asthma; Health Care Costs; Homeostasis; Human; Hyperplasia; In Vitro; Inflammation; Inflammatory; Injury; Interleukin-10; Interleukin-13; Mediating; Mediator of activation protein; Metaplasia; Metaplastic; Microscopy; Modality; Molecular; Mucolytics; Mucous body substance; Mus; Musculoskeletal; Nature; Ovalbumin; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase II Clinical Trials; Phenotype; Play; Population; Process; Production; Proliferating; Proteins; Pyroglyphidae; Reporting; Resolution; Respiratory physiology; Rho-associated kinase; Role; STAT1 gene; STAT6 Transcription Factor; Seminal; Signal Pathway; Signal Transduction; Sphingosine; Structure of parenchyma of lung; Symptoms; Testing; Therapeutic; Time; Tissues; Video Microscopy; airway hyperresponsiveness; airway obstruction; asthmatic; base; cancer therapy; cell fixing; cytokine; cytotoxic; disorder prevention; efficacy testing; inhibitor/antagonist; insight; interest; live cell imaging; mimetics; monolayer; mouse model; mucus hypersecretion; novel; novel strategies; polarized cell; preclinical trial; programs; prophylactic; response; restoration; screening; small hairpin RNA; small molecule; small molecule inhibitor; small molecule therapeutics; sphingosine kinase; targeted treatment; therapeutic target; tissue fixing; transcription factor; two-photon

Project Summary: Allergic asthma is a chronic inflammatory disease that directly affects large American population with an estimated total annual healthcare cost of 20 billion dollars. The heterogeneous nature driven by different underlying disease processes makes it challenging to treat and control the asthma symptoms. Therefore, in addition to having the effective disease prevention and management strategies, the development of novel and phenotype-specific therapeutic approaches is of great interest. Interleukin (IL)-13 has been recognized as a central mediator of TH2-dominant asthmatic phenotypes including excess mucus production and airway hyperresponsiveness. Consequently, therapeutics targeting IL-13 are very attractive with studies already in phase II clinical trials. But initial studies reported some adverse events associated with the use of IL-13 targeting therapeutics because IL-13 also elicits other immunoregulatory effects. For example, IL-13 suppresses TH17 cytokine production in an IL-10-dependent manner and thereby may play an important role in TH17-associated autoimmune diseases. Accordingly, patients receiving IL-13–targeted therapeutics had significantly increased adverse events involving the musculoskeletal and autoimmune-related diseases. Therefore, identifying targets that specifically affect the proliferative and metaplastic activity of IL-13 is crucial. The current study proposes a novel approach that switches the proliferative IL-13 to one that causes cell death in airway epithelial cells. IL-13 exerts proliferative effect by modulating various programmed cell death pathways. The anti-apoptotic protein, Bcl-2 inhibits apoptosis directly by interacting with various pro-apoptotic BH3-domain effector proteins. Indeed, our preliminary studies showed that IL-13 induced the proliferation and sustained the hyperplastic mucous cells by upregulating Bcl-2 expression. Blocking Bcl-2 by shRNA-based approach and by a small molecule Bcl-2 antagonist, ABT-263 reduced the mucus expression and caused mucous cell death. Screening for the pro-apoptotic factors showed that IL-13 induced a BH3-domain only protein, Bik, an epithelial cell death regulator in a STAT1-dependent manner. Thus, IL-13 modulates both Bcl-2 and Bik, the anti- and pro- cell death proteins, respectively to alter the cell fate, and blocking Bcl-2 results in a Bik-mediated cell death. Accordingly, the lung tissue sections from ABT-263 treated mice showed apically elevated apoptotic epithelial cells suggesting that epithelial extrusion or a programmed elimination might be involved in the removal of dying cells. In addition, the in-vitro studies with induced expression of Bik selectively removed the dying cells by extrusion with a characteristic actinomyosin ring formation. Therefore, we propose to investigate the cell death and extrusion process of airway epithelial cells by fixed- and live-cell imaging, and to interrogate the signaling pathways involved. We propose to test our findings in-vitro in both monolayer and differentiated culture settings, and in animal models. The efficacy of inhaled ABT-263 in regulating hyperplastic mucous cells will be tested in relevant mouse models of allergic asthma. More importantly, therapeutics for controlling mucous hypersecretion are limited with only few potential drugs currently in preclinical and clinical trials, therefore targeting hyperplastic mucous cells could represent a new class of mucolytic therapies. The use of ABT-263, an orally bioavailable Bcl-2 inhibitor that is currently being tested for human cancer treatment could help expedite its efficacy testing in asthmatics with debilitating mucous phenotype. In addition, the proposed studies will help develop a comprehensive understanding about the molecular mechanisms by which blocking Bcl-2 facilitates the IL-13–mediated mucous cell death. The long term objective of these studies is to help understand the pathogenesis of mucus hypersecretion observed in chronic airway diseases.

项目概要： 过敏性哮喘是一种慢性炎症性疾病，直接影响大量美国人口， 估计每年的医疗费用为200亿美元。由不同驱动的异质性 潜在的疾病过程使得治疗和控制哮喘症状具有挑战性。因此在 除了有效的疾病预防和管理策略外， 表型特异性治疗方法引起了极大的兴趣。白细胞介素（IL）-13已被认为是一种 TH 2-显性哮喘表型的中枢介质，包括过度粘液产生和气道 高反应性因此，靶向IL-13的治疗剂在已经进行的研究中是非常有吸引力的。 II期临床试验。但最初的研究报告了一些与使用IL-13相关的不良事件 因为IL-13还具有其他免疫调节作用。例如，IL-13 以IL-10依赖的方式抑制TH 17细胞因子的产生，从而在 TH 17相关的自身免疫性疾病。因此，接受IL-13靶向治疗的患者 涉及肌肉骨骼和自身免疫相关疾病的不良事件显著增加。 因此，鉴定特异性影响IL-13增殖和化生活性的靶点至关重要。 目前的研究提出了一种新的方法，将增殖性IL-13转换为导致细胞死亡的IL-13 呼吸道上皮细胞。IL-13通过调节各种程序性细胞死亡发挥增殖作用 途径。抗凋亡蛋白Bcl-2通过与多种促凋亡蛋白相互作用直接抑制细胞凋亡。 BH 3结构域效应蛋白。事实上，我们的初步研究表明，IL-13诱导增殖， 通过上调Bcl-2的表达来维持增生的粘液细胞。基于shRNA的Bcl-2阻断剂 通过小分子Bcl-2拮抗剂，ABT-263减少了粘液表达， 粘液细胞死亡筛选促凋亡因子显示IL-13仅诱导BH 3结构域 Bik蛋白是一种STAT 1依赖性的上皮细胞死亡调节因子。因此，IL-13既调节Bcl-2， 和Bik，抗细胞死亡蛋白和促细胞死亡蛋白，分别改变细胞命运，阻断Bcl-2导致细胞凋亡。 Bik介导的细胞死亡。因此，来自ABT-263处理的小鼠的肺组织切片显示顶部 凋亡上皮细胞增多，提示上皮细胞挤出或程序性消除可能是 参与了死亡细胞的清除此外，体外研究中，Bik的选择性诱导表达 通过挤压去除垂死的细胞，并形成特征性的肌动蛋白环。所以我们提出 通过固定和活细胞成像研究气道上皮细胞的细胞死亡和挤出过程， 来研究相关的信号通路我们建议在体外单层和 差异化的培养环境和动物模型。ABT-263吸入治疗乳腺癌的疗效观察 将在相关的过敏性哮喘小鼠模型中测试粘液细胞。更重要的是， 控制粘液分泌过多受到限制，目前临床前和临床上仅有少数潜在药物 因此，靶向增生性粘液细胞的试验可能代表一类新的粘液溶解疗法。的 使用ABT-263，一种口服生物可利用的Bcl-2抑制剂，目前正在测试用于人类癌症治疗 可以帮助加快其在具有衰弱粘液表型的哮喘患者中的疗效测试。此外该 拟议的研究将有助于全面了解分子机制， 阻断Bcl-2促进IL-13介导的粘液细胞死亡。这些研究的长期目标是 有助于了解慢性气道疾病中粘液分泌过多的发病机制。