An integrated mass spectrometry approach to study heparin structure-bioactivity

研究肝素结构-生物活性的综合质谱方法

基本信息

批准号：
10531619
负责人：
IGOR A KALTASHOV
金额：
$ 42.25万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10531619
关键词：
Address Affect Affinity Allosteric Regulation Alpha Granule Antibodies Anticoagulants Architecture Behavior Binding Binding Proteins Biopolymers Cell Differentiation process Cell Proliferation Collaborations Complex Development Diagnosis Diagnostic Electrons Environment Etiology Exhibits Exopeptidase Glycosaminoglycans Health Hematology Heparin Heparin Binding Heterogeneity Human body Immune System Diseases Intervention Knowledge Maps Mass Spectrum Analysis Measures Medical Medicine Molecular PF4 Gene Pathogenicity Pathology Patients Physiological Processes Plasma Play Predisposition Process Prognosis Protein Isoforms Proteins Proteolytic Processing Recombinant Antibody Research Resolution Role Site Source Structure System Techniques Therapeutic Therapeutic Agents Therapeutic Intervention Thrombin Thrombocytopenia Thrombosis Universities Work analytical tool cohort design diagnostic tool granule cell heparin-induced thrombocytopenia immunogenic immunogenicity immunoregulation macromolecule mast cell molecular modeling molecular targeted therapies patient subsets prevent prognostic tool small molecule stoichiometry structural determinants success therapeutic target therapeutically effective tool volunteer

项目摘要

PROJECT SUMMARY Heparin is arguably the most versatile biopolymer, capable of interacting with and modulating the behavior of an impressive variety of biomolecules even outside of its native environment (mast cell granules). A large fraction of heparin interactome in the human body are key players in a variety of physiological processes (including several pathologies) and are considered high-value therapeutic targets, although at present antithrombin remains the only protein whose interaction with heparin has been successfully exploited in medicine. Successful exploitation of heparin’s unique versatility for other therapeutic purposes critically depends on the ability to characterize its interactions with relevant proteins; however, specific molecular mechanisms remain elusive outside of the very few extensively studied systems. In the previous period of support, we developed powerful analytical tools capable of providing information on protein/heparin interactions at the whole molecule level, as well as identifying specific structural motifs within highly heterogeneous heparin macromolecules that enable their association with specific proteins. Building upon this success, we now propose to focus our inquiry on understanding the molecular mechanisms underlying etiology of heparin-induced thrombocytopenia (HIT), a serious (and potentially fatal) immune disorder affecting up to 5% of patients receiving heparin as an anticoagulant. Despite the central role played by heparin in formation of antigenic aggregates that may lead to development of HIT, relatively little is known about the specific molecular mechanisms governing these interactions. This not only creates a tremendous challenge vis-a-vis the ability to design efficient anti-HIT therapeutic strategies, but also prevents a reliable prognosis of the occurrence of this pathology. Recognizing the importance of multiple factors that may modulate interaction of heparin with relevant proteins (mostly platelet factor 4, PF4), we will use a multi-level strategy to study the mechanism of PF4/heparin association, architecture of these aggregates and specific features that make them immunogenic and trigger the onset of HIT. The work will be carried out in close collaboration with the Hematology team at McMaster University Health Centre headed by Dr. I. Nazy, a leading expert in the field of thrombocytopenia and thrombosis. This knowledge will catalyze efforts to develop reliable diagnostic and prognostic tools for HIT, and will be critical for designing safe and effective therapeutic intervention strategies.

项目摘要肝素可以说是最通用的生物聚合物，能够与令人印象深刻的生物分子甚至在其本地环境（肥大细胞颗粒）之外。一大部分在人体中肝素互动的组合是各种物理过程中的关键参与者（包括几种病理），被认为是高价值治疗靶标，尽管目前抗凝血酶仍然是唯一在医学上成功利用与肝素相互作用的蛋白质。针对其他治疗目的，对肝素独特的多功能性的开发在很大程度上取决于能力表征其与相关蛋白质的相互作用；但是，特定的分子机制仍然难以捉摸在几个广泛研究的系统之外。在上一个阶段，我们发展了强大的能够在整个分子水平上提供有关蛋白质/肝素相互作用的信息的分析工具，以及确定高度异质肝素大分子内的特定结构基序它们与特定蛋白质的关联。在这一成功的基础上，我们现在建议将我们的询问集中在了解肝素诱导的血小板减少症（HIT）的病因学分子机制，A 严重（可能是致命的）免疫疗法，多达5％接受肝素的患者抗凝物。尽管肝素在形成抗原骨料中的核心作用可能导致命中的发展，对这些的特定分子机制知之甚少互动。对于设计有效的反击的能力，这不仅带来了巨大的挑战治疗策略，但也可以防止这种病理发生的可靠预后。认可多种因素的重要性可能调节肝素与相关蛋白的相互作用（主要是血小板因素4，PF4），我们将使用多层策略来研究PF4/肝素协会的机制在这些聚集体和特定特征中，它们使它们免疫原性并触发了命中的发作。工作将与麦克马斯特大学健康中心的血液学团队密切合作进行由血小板减少症和血栓形成领域的领先专家I. Nazy博士。这些知识将催化努力开发可靠的诊断和预后工具以进行命中，并且对于设计安全和设计至关重要有效的治疗干预策略。