Exploiting Siglec-6 for targeted anti-allergy drug delivery into human mast cells

利用 Siglec-6 将靶向抗过敏药物输送到人体肥大细胞中

基本信息

批准号：
10194041
负责人：
Bruce S Bochner
金额：
$ 18.67万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-08 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10194041
关键词：
Affect Agammaglobulinaemia tyrosine kinase Allergens Allergic Allergic Disease Allergic Reaction Allergic inflammation Allergic rhinitis American Anaphylaxis Animal Model Anti-Allergic Agents Asthma Atopic Dermatitis Basophils Binding Biodistribution Biological Assay Bolus Infusion Cell Line Cell Surface Proteins Cell Surface Receptors Cell secretion Cells Data Detection Development Dose Drug Carriers Drug Controls Drug Delivery Systems Drug Targeting Emergency Situation Encapsulated Endocytosis Engineering Epinephrine FDA approved Food Hypersensitivity Genomics Goals Grant Hospitals Human Hypersensitivity IgE IgE Receptors Immunoglobulins In Vitro Inductively Coupled Plasma Mass Spectrometry Intervention Irrigation Isotope Labeling Isotopes Kinetics Knock-in Knock-in Mouse Label Lectin Life Liquid Chromatography Malignant Neoplasms Mediating Methods Micelles Modeling Modification Monoclonal Antibodies Mus Nanostructures Nature Oral Organ Pharmaceutical Preparations Pharmacology Pharmacotherapy Polymers Premedication Proteomics Protocols documentation Publishing Reaction Reagent Receptor Signaling Research Safety Sampling Severities Sialic Acids Side Signal Transduction Specificity Structure Sulfur Surface System Testing Therapeutic Therapeutic Agents Thulium Tissue Sample Tissues Tyrosine Kinase Inhibitor Validation Visit Wild Type Mouse antibody conjugate cell behavior cost density desensitization design drug testing experimental group humanized mouse improved in vivo kinase inhibitor lipophilicity mast cell mouse model nanocarrier nanomolar nanoscale novel prevent prophylactic protective effect receptor binding receptor mediated endocytosis sialic acid binding Ig-like lectin side effect tandem mass spectrometry targeted delivery targeted treatment tool uptake

项目摘要

Project Summary Allergic diseases are IgE-dependent, mast cell-mediated conditions that affect more than 50 million Americans. Severe allergic reactions result in anaphylaxis and can be potentially life threatening, sometimes requiring hospital visits in conjunction with emergency interventions such as the use of epinephrine autoinjectors. Various pharmacotherapies have been developed for the treatment of allergic rhinitis, atopic dermatitis, asthma and food allergy, but none are capable of directly regulating mast cell behavior in a way that truly prevents mast cell reactivity to allergens. A therapeutic strategy is therefore needed to 1) selectively target mast cells for delivery of 2) a potent drug that can directly prevent IgE-dependent mast cell secretion. Sialic acid binding immunoglobulin-like lectin 6 (Siglec-6) is a cell surface receptor that is highly and preferentially expressed on all types of mast cells. While its specific function and associated mechanism of signaling is largely unexplored, its homology to other mast cell Siglecs, like CD33, Siglec-7 and Siglec-8, makes it likely to be inhibitory in function. Separate from its canonical signaling function, engagement of Siglec-6 results in its endocytosis. This supports the hypothesis that Siglec-6 can be exploited for targeted drug delivery into mast cells. Development of a Siglec-6-targeted therapy requires an appropriate delivery vehicle. Previously, the Scott Lab has developed and tested a variety of stable, scalable and customizable drug carriers that each possess distinct structure-dependent advantages for controlled delivery. For example, spherical and filamentous nanocarriers can both transport lipophilic drugs, but their differences in aspect ratio result in distinct capacity for receptor-mediated endocytosis and signaling when targeting moieties are presented on their surfaces. In terms of therapeutic agents, Acalabrutinib (AcB) and other Bruton’s kinase inhibitors (BTKi) irreversibly bind and inhibit BTK to prevent IgE-mediated activation of mast cells via FcεRI, a key driver of anaphylaxis. The Bochner Lab has just demonstrated that pretreatment with AcB in a novel humanized mast cell mouse model of anaphylaxis has profound protective effects regarding anaphylaxis severity and improved survival. In parallel, BTKi completely shut off IgE receptor signaling in both human basophils and mast cells in vitro, with IC50’s in the low to mid nanomolar range. However, the pleiotropic nature of systemic BTKi treatment results in potentially serious side effects, and thus BTKi are not approved for use outside of cancer indications. To test the hypothesis that Siglec-6-mediated targeted delivery of AcB will result in specific inhibition of allergic mast cell activation, two Specific Aims will be achieved: (1) to optimize and characterize the binding kinetics, specificity, and inhibitory activity of Siglec-6/BTKi loaded micellar and filamentous nanocarriers; and (2) to compare the specificity and inhibitory effects of these nanocarriers in Siglec-6 mast cell knock-in mice and humanized mast cell mice. The overarching goal is to generate proof-of-concept data for a subsequent R01 application to more fully develop and test drug delivery systems that selectively deliver drugs into mast cells.

项目摘要过敏性疾病是IgE依赖性的，肥大细胞介导的疾病，影响超过5000万美国人。严重的过敏反应会导致过敏反应，并可能危及生命，有时需要与紧急干预措施（例如使用肾上腺素自动注射器）一起就诊。已经开发出各种药物疗法用于治疗过敏性鼻炎，特应性皮炎，哮喘和食物过敏细胞对过敏原的反应。因此，需要一种治疗策略1）选择性靶向肥大细胞进行输送 2）可以直接防止IgE依赖性肥大细胞分泌的潜在药物。唾液酸结合免疫球蛋白样讲座6（SIGLEC-6）是一种细胞表面受体，高度高，并且优先在所有类型的肥大细胞上表达。而其特定功能和相关机制信号传导在很大程度上是出乎意料的，它与其他肥大细胞siglecs（如CD33，Siglec-7和Siglec-8）的同源性使它可能会抑制功能。与其规范信号函数分开，SIGLEC-6结果的参与度在其内吞作用中。这支持了可以探索Siglec-6的假设肥大细胞。 SIGLEC-6靶向疗法的开发需要适当的送货工具。以前，斯科特实验室已经开发并测试了各种稳定，可扩展和可定制的药物载体具有不同的结构依赖性优势来控制交付。例如，球形和丝状纳米载体既可以运输亲脂性药物，但是它们的纵横比的差异使得当靶向部分时，受体介导的内吞和信号传导在其表面上呈现。在治疗剂方面，Acalabrutinib（ACB）和其他Bruton的激酶抑制剂（BTKI）不可逆转地结合并抑制BTK，以防止IgE介导的肥大细胞通过过敏反应的关键驱动器FcεRI激活。这 Bochner Lab刚刚证明，在新型的人源肥大细胞小鼠模型中对ACB进行预处理过敏反应对过敏反应和改善生存具有深远的保护作用。并联， BTKI在体外完全关闭了人类嗜碱性粒细胞和肥大细胞中的IgE接收器信号，其中IC50在低至中纳摩尔范围。但是，全身BTKI治疗的多效性可能导致严重的副作用，因此BTKI未被批准在癌症适应症之外使用。为了测试SIGLEC-6介导的ACB靶向递送的假设将导致特定的抑制过敏性肥大细胞激活，将实现两个具体目标：（1）优化和表征结合 SIGLEC-6/BTKI装载的胶束和丝状纳米载体的动力学，特异性和抑制活性；（2）比较这些纳米载体在SIGLEC-6肥大细胞敲入小鼠中的特异性和抑制作用人源化肥大细胞小鼠。总体目标是为随后的R01生成概念验证数据应用于更充分的开发和测试药物输送系统，这些药物有选择地将药物输送到肥大细胞中。