Eosinophils and Eosinophil Activation in the Pathogenesis of Human Disease

人类疾病发病机制中的嗜酸性粒细胞和嗜酸性粒细胞激活

• 批准号: 10692147
• 负责人: Amy Klion
• 金额: $ 182.38万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10692147
• 关键词: Address; Affect; Aftercare; Allergens; Allergic; Angioneurotic Edema; Automobile Driving; Barbering; Benign; Biological; Biological Assay; Biological Markers; Blood; COVID-19; Cells; Chemotaxis; Clinical; Collaborations; Cytolysis; Data; Disease; Disseminated eosinophilic collagen disease; Dose; Effector Cell; Enrollment; Eosinophil Granule Proteins; Eosinophil-Derived Neurotoxin; Eosinophilia; Eosinophilic leukemia; Epithelial; Filariasis; Follow-Up Studies; Goals; Homeostasis; Human; Hypersensitivity; Imatinib; Immune response; In Vitro; Infection; Innate Immune Response; Intervention; Kinetics; Label; Laboratories; Ligands; Longterm Follow-up; Lung; Lung infections; Macaca; Macaca mulatta; Measurement; Measures; Mediating; Methods; Modeling; Multicenter Studies; Mus; Mycobacterium Infections; Mycobacterium tuberculosis; Natural Killer Cells; Organ; PDGFRA gene; Pathogenesis; Patients; Peripheral Blood Eosinophilia; Phase; Plasma; Prednisone; Rare Diseases; Reaction; Refractory; Research Design; Residual state; Role; Safety; Serum; Site; Steroids; Structure of parenchyma of lung; Symptoms; Therapeutic Agents; Tissues; Toxic effect; Tuberculosis; Tyrosine Kinase Inhibitor; Urine; Wegener' s Granulomatosis; Zebrafish; clinical subtypes; cohort; conventional therapy; diagnostic tool; drug action; effective therapy; eosinophil; eosinophilic asthma; gastrointestinal; helminth infection; human disease; human tissue; in vivo; inhibitor; insight; interest; investigator-initiated trial; mast cell; mepolizumab; microbial; migration; mouse model; multiplex assay; neoplastic; neutrophil; novel; novel diagnostics; oxysterol binding protein; pathogen; patient subsets; phase 3 study; phase III trial; placebo controlled study; predicting response; recruit; response; targeted treatment

Using a cohort of more than 600 subjects with a wide variety of eosinophilic disorders, ranging from benign eosinophilia to eosinophilic leukemia, we have continued to identify and characterize novel subgroups of patients with eosinophilia and to explore responses to targeted therapies with the goal of increasing our understanding of the role of eosinophils in homeostasis and disease pathogenesis. We have also expanded our collaborative studies examining the role of eosinophils in the immune response to microbial infections, including M. tuberculosis, and the role of eosinophils in the clinical manifestations of COVID19 (Espinoza et al.J Allergy Clin Immunol Pract 2022). One of the difficulties in assessing responses to clinical interventions in HES is the lack of specific measures of eosinophil-mediated disease activity. This is particularly problematic in the setting of eosinophil-depleting therapies that may lower the eosinophil count despite clinical evidence of continuing eosinophil-mediated pathogenesis. We have previously described a multiplex assay to measure eosinophil granule proteins in the blood and shown a correlation between other markers of eosinophil activation and serum levels of eosinophil-derived neurotoxin (EDN) that is independent of the blood eosinophil count (Makiya et al. J Immunol Methods 2014). In the past year, we explored the use of this assay to measure eosinophil granule proteins in the blood and urine of patients with hypereosinophilic syndrome (HES), eosinophilic granulomatosis with polyangiitis (EGPA), and eosinophilic asthma before and after treatment with eosinophil-depleting therapies (Makiya et al. Allergy 2022). Whereas both plasma and urine measurement of EDN were elevated in patients with active disease and decreased with effective therapy, unlike plasma measurement, urine measurement is non-invasive and unaffected by cellular lysis. Moreover, the lack of correlation between AEC or prednisone dose and urine but not plasma EDN levels suggests that measurement of urine EDN may be a better biomarker of disease activity in patients with eosinophilic disorders. Targeted therapies not only provide new options for patients but provide insight into the effects of eosinophil depletion in humans and the mechanisms driving eosinophilia and eosinophil activation in HES. Ongoing studies include the use of tyrosine kinase inhibitors (imatinib and ruxolitinib) for the treatment of steroid-refractory HES, a single site study of mepolizumab for the treatment of episodic angioedema and eosinophilia, a multicenter phase 3 trial of benralizumab for HES, as well as long-term followup studies of HES patients enrolled on investigator-initiated trials of benralizumab and dexpramipexole. We have previously described the results of our single center phase 2 placebo-controlled study of benralizumab that demonstrated efficacy of this agent in patients with treatment-refractory PDGFRA-negative HES (Kuang et al. N Engl J Med 2019). These results were instrumental in the initiation of an ongoing multicenter phase 3 study of benralizumab in HES and provided support for the safety of eosinophil-depletion in humans (Manetz et al.J Allergy Clin Immunol Pract 2020). Detailed analysis of the clinical, laboratory and histochemical findings in the 7 patients with gastrointestinal involvement suggest that residual symptoms in some patients despite complete depletion of eosinophils in the blood and tissue may reflect persistent epithelial changes and/or mast cell persistence (Kuang et al. J Allergy Clin Immunol Pract 2022). As the number of therapeutic agents that affect eosinophils, eosinophil migration and eosinophil activation increase, it is becoming increasingly important to identify predictors of response. We have previously shown that the subtype of HES is an important predictor of response to conventional and targeted therapies (Kuang et al. J Allergy Clin Immunol Pract 2018; Khoury et al. J Allergy Clin Immunol Pract 2018). A recent multicenter study of off-label biologic use for the treatment of HES confirmed variable responsiveness to the different agents due, at least in part, to HES clinical subtype (Chen et al. J Allergy Clin Immunol Pract 2022). Other factors, including the mechanism of action of the drug and the role of other cells in disease pathogenesis are clearly also important both in determining efficacy and potential toxicity in patients with eosinophilic disorders. The latter is exemplified by the effect of dupilumab on blood eosinophilia (Wechsler et al. J Allergy Clin Immunol Pract 2022). Historically, eosinophils have been viewed as effector cells with primary roles in the response to helminth infection and allergens. Consistent with this, recent studies in our group have focused on the role of eosinophils and eosinophil activation in the clinical manifestations and post-treatment reactions in filariasis (Herrick et al. Clin Infect Dis 2020; Legrand et al. Clin Infect Dis 2017; Legrand et al. Clin Infect Dis 2021). Over the past decade, however, there has been increasing interest in the role of eosinophils in the modulation of responses to non-helminth pathogens. To address this issue, we initiated a collaboration with Dr. Katrin Mayer-Barber to examine the role of eosinophils in the host immune response to pulmonary infection with Mycobacterium tuberculosis (Mtb). After initial studies in murine models demonstrated that the early influx of neutrophils and NK cells into the lung post-Mtb infection is eosinophil-dependent, suggesting that eosinophils may be responsible for orchestrating the pulmonary innate immune response after Mtb infection, these findings were confirmed using rhesus macaque and zebrafish models of mycobacterial infection and human tissues from patients with tuberculosis (Bohrer et al. J Exp Med 2021). In a more recent study designed to explore the mechanism and kinetics of eosinophil recruitment to the lung in Mtb infection, we used macaque and murine models to demonstrate that eosinophils are recruited to the infected lung within 1-2 weeks and interact with Mtb infected cells in the airways (Bohrer et al. Cell Rep 2022). In mice, this migration was CCR3-independent but required expression of GPR183, an oxysterol receptor present on human blood eosinophils and human eosinophils isolated from Mtb-infected lung tissue. Consistent with the in vivo murine data, in vitro human eosinophil chemotaxis to oxysterol ligands was abrogated by specific GPR183 inhibitors. These data support an early role for eosinophils in the protective immune response to Mtb infection.

通过使用 600 多名患有各种嗜酸性粒细胞疾病（从良性嗜酸性粒细胞增多症到嗜酸性粒细胞白血病）的受试者队列，我们​​继续识别和表征新的嗜酸性粒细胞增多症患者亚组，并探索对靶向治疗的反应，目的是提高我们的治疗效果。了解嗜酸性粒细胞在体内平衡和疾病发病机制中的作用。我们还扩大了合作研究，检查嗜酸性粒细胞在微生物感染（包括结核分枝杆菌）的免疫反应中的作用，以及嗜酸性粒细胞在 COVID19 临床表现中的作用（Espinoza 等人，J Allergy Clin Immunol Pract 2022）。 评估 HES 临床干预反应的困难之一是缺乏嗜酸性粒细胞介导的疾病活动的具体措施。尽管临床证据表明嗜酸性粒细胞介导的发病机制持续存在，但在嗜酸性粒细胞耗竭疗法的情况下，这尤其成问题，该疗法可能会降低嗜酸性粒细胞计数。我们之前描述了一种测量血液中嗜酸性粒细胞颗粒蛋白的多重测定法，并显示了嗜酸性粒细胞活化的其他标志物与嗜酸性粒细胞衍生神经毒素（EDN）的血清水平之间的相关性，该相关性与血液中嗜酸性粒细胞计数无关（Makiya等人，J）免疫方法 2014）。在过去的一年中，我们探索了使用这种测定法来测量嗜酸性粒细胞增多综合征（HES）、嗜酸性肉芽肿性多血管炎（EGPA）和嗜酸性粒细胞哮喘患者在接受嗜酸性粒细胞耗竭治疗前后的血液和尿液中的嗜酸性粒细胞颗粒蛋白。疗法（Makiya et al. Allergy 2022）。与血浆测量不同，血浆和尿液中 EDN 测量结果在患有活动性疾病的患者中均升高，并随着有效治疗而降低，而尿液测量是非侵入性的，不受细胞裂解的影响。此外，AEC 或泼尼松剂量与尿液而非血浆 EDN 水平之间缺乏相关性，这表明尿液 EDN 的测量可能是嗜酸性粒细胞疾病患者疾病活动性的更好生物标志物。 靶向治疗不仅为患者提供了新的选择，而且还提供了对人类嗜酸性粒细胞耗竭的影响以及 HES 中嗜酸性粒细胞增多和嗜酸性粒细胞激活的驱动机制的深入了解。正在进行的研究包括使用酪氨酸激酶抑制剂（伊马替尼和鲁索替尼）治疗类固醇难治性 HES、美泊利单抗治疗阵发性血管性水肿和嗜酸性粒细胞增多症的单中心研究、贝那利珠单抗治疗 HES 的多中心 3 期试验以及作为对参加研究者发起的贝那利珠单抗试验的 HES 患者的长期随访研究右旋普拉克索。我们之前已经描述了贝那利珠单抗的单中心 2 期安慰剂对照研究的结果，该研究证明了该药物对难治性 PDGFRA 阴性 HES 患者的疗效（Kuang 等人，N Engl J Med 2019）。这些结果有助于启动正在进行的贝那利珠单抗治疗 HES 的多中心 3 期研究，并为人类嗜酸性粒细胞耗竭的安全性提供支持（Manetz 等人，J Allergy Clin Immunol Pract 2020）。对 7 例胃肠道受累患者的临床、实验室和组织化学结果的详细分析表明，尽管血液和组织中的嗜酸性粒细胞完全耗尽，一些患者的残留症状可能反映了持续的上皮变化和/或肥大细胞的持续存在（Kuang 等，2017）。 J 过敏临床免疫实践杂志 2022）。 随着影响嗜酸性粒细胞、嗜酸性粒细胞迁移和嗜酸性粒细胞活化的治疗药物数量的增加，识别反应的预测因子变得越来越重要。我们之前已经证明，HES 亚型是对常规和靶向治疗反应的重要预测因子（Kuang 等人，J Allergy Clin Immunol Pract 2018；Khoury 等人，J Allergy Clin Immunol Pract 2018）。最近一项关于标签外生物制剂治疗 HES 的多中心研究证实，至少部分由于 HES 临床亚型而对不同药物产生不同的反应（Chen 等人，J Allergy Clin Immunol Pract 2022）。其他因素，包括药物的作用机制和其他细胞在疾病发病机制中的作用，显然对于确定嗜酸性粒细胞疾病患者的疗效和潜在毒性也很重要。后者的例子是 dupilumab 对血液嗜酸性粒细胞增多的影响（Wechsler 等人，J Allergy Clin Immunol Pract 2022）。 从历史上看，嗜酸性粒细胞一直被视为在响应蠕虫感染和过敏原中起主要作用的效应细胞。与此相一致的是，我们小组最近的研究重点是嗜酸性粒细胞和嗜酸性粒细胞活化在丝虫病临床表现和治疗后反应中的作用（Herrick et al. Clin Infect Dis 2020；Legrand et al. Clin Infect Dis 2017；Legrand等人，临床感染疾病 2021）。然而，在过去的十年中，人们对嗜酸性粒细胞在调节非蠕虫病原体反应中的作用越来越感兴趣。为了解决这个问题，我们与 Katrin Mayer-Barber 博士合作，研究嗜酸性粒细胞在宿主对结核分枝杆菌 (Mtb) 肺部感染的免疫反应中的作用。 在小鼠模型中的初步研究表明，Mtb 感染后中性粒细胞和 NK 细胞早期流入肺部是嗜酸性粒细胞依赖性的，这表明嗜酸性粒细胞可能负责协调 Mtb 感染后的肺部先天免疫反应，这些发现得到了证实分枝杆菌感染的恒河猴和斑马鱼模型以及结核病患者的人体组织（Bohrer 等人，J Exp Med 2021）。在一项旨在探索 Mtb 感染中嗜酸性粒细胞募集到肺部的机制和动力学的最新研究中，我们使用猕猴和小鼠模型来证明，嗜酸性粒细胞在 1-2 周内募集到受感染的肺部，并与 Mtb 感染的细胞相互作用。气道（Bohrer 等人。Cell Rep 2022）。在小鼠中，这种迁移不依赖于 CCR3，但需要 GPR183 的表达，GPR183 是一种存在于人血液嗜酸性粒细胞和从 Mtb 感染的肺组织中分离的人嗜酸性粒细胞上的氧甾醇受体。与体内小鼠数据一致，体外人嗜酸性粒细胞对氧甾醇配体的趋化性被特定的 GPR183 抑制剂消除。这些数据支持嗜酸性粒细胞在针对结核分枝杆菌感染的保护性免疫反应中的早期作用。