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

基本信息

批准号：
10636783
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
项目类别：
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

项目摘要

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理和宿主相互作用。这些蛋白质不是更常见的渠道，转运蛋白和GPCR，因此提供了有关的新知识膜蛋白结构，功能和动力学。细菌膜蛋白是抗生素的靶标哪种阻力是一个巨大的威胁。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性递送技术有吸引力（例如受体介导的摄取）。这 MIRA应用概述了我们最近在理解几种不同细菌膜方面的努力蛋白质以及富有成果的合作，将生物物理学桥接到不同的生物医学领域。 OPA蛋白来自淋病奈瑟氏菌和脑膜炎N.是外膜蛋白，与各种宿主受体结合诱导细菌吞没。这些受体中的几个在癌症中过表达，可能提供治疗分娩的目标。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针对人类细胞的靶向脂质体递送。我们已经开始了解OPA结构，并具有有关OPA与受体之间相互作用的初步数据 CEACAM1。此外，我们成功地创建了诱导受体介导的Opa-脂质体吞噬作用。未来的方向集中于一种多学科方法来理解的决定因素基于相互作用的Opa受体选择性和工程师的治疗递送脂质体。其他感兴趣的治疗递送功能是细胞跟踪和控制细胞命运。印加，来自衣原体，通过与宿主网罗互动允许细菌避免溶酶体的劫持劫持人口贩运降解。我们提出了多种生物物理和结构方法来理解结构 - 印加和与自身的互动和贪食的功能关系，以设计细胞内递送可以避免溶酶体降解的系统。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，以帮助抵抗耐药细菌的增加。信号肽酶 II，LSPA是一个潜在的靶标，因为它是在所有革兰氏阴性细菌而不是人类中发现的。环霉素被分离并在1978年鉴定出抗生素活性。尽管合成和结构是众所周知，环霉素尚未成为一种治疗性。我们旨在探索LSPA对LSPA的结合和抑制球霉素样肽是为了鉴定革兰氏阴性细菌的可行抗生素。结果的结果提案提供有关细菌膜蛋白及其在中的作用的独特知识和见解使用生物物理方法的发病机理，并将制定设计治疗治疗的策略细菌感染和各种涉及CEACAM受体的癌症。