Biophysical understanding of pathogen-host membrane protein interactions for drug discovery and delivery

对药物发现和递送的病原体-宿主膜蛋白相互作用的生物物理学理解

• 批准号: 9920747
• 负责人: Linda M Columbus
• 金额: $ 38.15万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9920747
• 关键词: Antibiotics; Bacteria; Bacterial Infections; Binding; Binding Proteins; Biophysics; CEACAM1; Cells; Collaborations; Data; Drug Delivery Systems; Engineering; Future; G-Protein-Coupled Receptors; Gram-Negative Bacteria; Human; Knowledge; Liposomes; Malignant Neoplasms; Mediating; Membrane Proteins; Neisseria gonorrhoeae; Neisseria meningitidis; Pathogenesis; Peptides; Phagocytosis; Proteins; Research; Resistance; Role; SNAP receptor; Structure; Structure-Activity Relationship; System; Technology; Therapeutic; bacterial resistance; base; biophysical techniques; combat; design; drug discovery; insight; interdisciplinary approach; interest; liposomal delivery; molecular recognition; novel; overexpression; pathogen; protein structure function; receptor; signal peptidase; targeted treatment; trafficking; uptake

Many membrane proteins mediate bacterial pathogenesis and host interactions. These proteins are not the more commonly investigated channels, transporters, and GPCRs and, therefore, provide new knowledge about membrane protein structure, function, and dynamics. Bacterial membrane proteins are targets of antibiotics for which resistance is a great threat. In addition, bacterial membrane proteins that interact with hosts have evolved functions that are attractive to therapeutic delivery technologies (e.g. receptor-mediated uptake). This MIRA application outlines our recent endeavors in understanding several different bacterial membrane proteins, as well as, fruitful collaborations bridging biophysics to different biomedical fields. Opa proteins from Neisseria gonorrhoeae and N. meningitidis are outer membrane proteins that bind to various host receptors that induce engulfment of the bacterium. Several of these receptors are overexpressed in cancers and may provide a target for therapeutic delivery. Knowledge of the structure, dynamics, and specific interactions of Opa proteins and receptors will be used to design targeted liposome delivery to human cells. We have begun to understand the Opa structure and have preliminary data on the interactions between Opa and the receptor CEACAM1. In addition, we have successfully created Opa-liposomes that induce receptor-mediated phagocytosis. Future directions focus on a multidisciplinary approach to understanding the determinants of Opa-receptor selectivity and engineer therapeutic delivery liposomes based on the interaction. Another function of interest to therapeutic delivery, is cellular tracking and controlling cellular fate. IncA, from Chlamydiae, hijacks host trafficking by interacting with host SNAREs allowing the bacterium to avoid lysosomal degradation. We propose a variety of biophysical and structural approaches to understanding the structure- function relationship of IncA and interactions with itself and SNAREs in order to design intracellular delivery systems that can avoid lysosomal degradation. Distinct from our research with Opa and IncA, we have begun to investigate potential antibiotic targets to help combat the increase in resistant bacteria. The signal peptidase II, LspA, is a potential target because it is found in all Gram-negative bacteria and not humans. Globomycin was isolated and the antibiotic activity was identified in 1978. Although the synthesis and structure are now known, globomycin has not become a therapeutic. We aim to explore the binding and inhibition of LspA with globomycin-like peptides in order to identify viable antibiotics for Gram-negative bacteria. The results of this proposal with provide unique knowledge and insights about bacterial membrane proteins and their roles in pathogenesis using biophysical approaches and will develop strategies for the design of therapeutics to treat bacterial infections and a variety of human cancers involving CEACAM receptors.

许多膜蛋白介导细菌发病机制和宿主相互作用。这些蛋白质不是 更常见的研究通道、转运蛋白和 GPCR，因此提供了关于 膜蛋白的结构、功能和动力学。细菌膜蛋白是抗生素的靶点 这种抵抗是一个巨大的威胁。此外，与宿主相互作用的细菌膜蛋白具有 进化的功能对治疗传递技术有吸引力（例如受体介导的摄取）。这 MIRA 应用概述了我们最近在了解几种不同细菌膜方面所做的努力 蛋白质，以及将生物物理学与不同生物医学领域联系起来的富有成效的合作。欧巴蛋白来自 淋病奈瑟菌和脑膜炎奈瑟菌是与各种宿主受体结合的外膜蛋白 诱导细菌的吞噬。其中一些受体在癌症中过度表达，并且可能 提供治疗递送的目标。了解 Opa 的结构、动力学和特定相互作用 蛋白质和受体将用于设计向人体细胞递送的靶向脂质体。我们已经开始 了解 Opa 结构并获得 Opa 与受体之间相互作用的初步数据 CEACAM1。此外，我们还成功创建了 Opa 脂质体，可诱导受体介导的 吞噬作用。未来的方向侧重于采用多学科方法来理解的决定因素 Opa 受体选择性并根据相互作用设计治疗递送脂质体。其他 治疗传递的重要功能是细胞跟踪和控制细胞命运。公司，来自 衣原体通过与宿主陷阱相互作用来劫持宿主贩运，从而使细菌避免溶酶体 降解。我们提出了多种生物物理和结构方法来理解结构- IncA 的功能关系以及与其自身和 SNARE 的相互作用，以便设计细胞内递送 可以避免溶酶体降解的系统。与我们与 Opa 和 IncA 的研究不同，我们已经开始 研究潜在的抗生素靶标，以帮助对抗耐药细菌的增加。信号肽酶 II，LspA，是一个潜在的目标，因为它存在于所有革兰氏阴性细菌中，而不是人类中。球霉素 1978年被分离出来，并鉴定了其抗生素活性。虽然现在其合成和结构已不明确 已知，球霉素尚未成为治疗药物。我们的目的是探索 LspA 的结合和抑制 球霉素样肽，以确定针对革兰氏阴性菌的可行抗生素。这样做的结果 提案提供了有关细菌膜蛋白及其在细菌中的作用的独特知识和见解 使用生物物理方法研究发病机制，并将制定治疗方法设计策略 涉及 CEACAM 受体的细菌感染和多种人类癌症。