Enzyme-Powered Self-Propelled DNA Nanoparticles for Disruption and Antibiotic Delivery in Topical Biofilms

用于局部生物膜破坏和抗生素递送的酶驱动自驱动 DNA 纳米颗粒

基本信息

批准号：
10528087
负责人：
Jeffrey Lawrence Moran
金额：
$ 18.02万
依托单位：
GEORGE MASON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-06 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10528087
关键词：
Acidic Region Animal Model Anti-Bacterial Agents Antibiotic Resistance Antibiotic Therapy Antibiotics Antigens Area Bacteria Biological Biological Assay Biomass Bladder Burn injury Bypass Caliber Ceftazidime Cessation of life Chemotaxis Clinical Clinical Trials Complex DNA DNA Maintenance Data Set Dependence Destinations Diffuse Disease Economics Environment Enzymes Extracellular Matrix Fibroblasts Fluorescence Spectroscopy Foundations Future Gastrointestinal Diseases Gel Health Care Costs Human Hydrogen Peroxide Hydrolase Immune response Infection Lead Location Locomotion Malignant Neoplasms Malignant neoplasm of urinary bladder Measures Mediating Medical Care Costs Methods Microbial Biofilms Microbiology Modeling Mucins Mus Nanotechnology Nutrient Output Patients Pattern Permeability Pharmaceutical Preparations Platinum Polymers Pseudomonas aeruginosa Shapes Skin Speed Stomach Stress Technology Testing Topical application Transmission Electron Microscopy Uncertainty Urea Urease Urine Work antimicrobial drug base biomaterial compatibility combinatorial design diabetic ulcer extracellular in vivo innovation mouse model multidisciplinary nanoparticle nanoscale open wound pH gradient particle pathogen quorum sensing remediation response skin lesion success targeted delivery

项目摘要

PROJECT SUMMARY/ABSTRACT Bacterial biofilms are responsible for most human infections, causing tens of thousands of deaths and billions in medical costs per year. Topical biofilms alone cause significant harm to patients by growing on open wounds, skin lesions, burn injuries, or diabetic ulcers, and elsewhere. Biofilms are notoriously difficult to eradicate, in large part because of the extracellular polymeric substance (EPS), a self-produced extracellular matrix in which biofilm bacteria reside. The EPS benefits bacteria in many ways, including mediating quorum sensing, providing nutrients, and blocking transport of antibiotics and host immune response. The ability to actively penetrate the EPS and deliver anti-bacterial cargo where it is most needed would bypass many of these protections and could thus have a transformative impact on the remediation of biofilms. First introduced in 2004, artificial self-propelled particles (SPPs) can propel themselves through complex biological media and deliver cargo to specific locations. Thus, SPPs hold significant potential for biomedical applications such as biofilm remediation. However, SPPs must overcome significant challenges in the form of biocompatibility, tracking, and control to be viable for clinical use. Here, we propose to leverage the burgeoning field of DNA nanotechnology to develop urease-powered DNA-origami-based self-propelled particles (DNA-SPPs) for biofilm remediation. As a model organism, we focus on the well-studied pathogen Pseudomonas aeruginosa. Aim 1 of this study will quantify the dependence of DNA-SPPs’ locomotion on local urea concentration and pH and elucidate the extent to which they perform chemotaxis in urea gradients. Aim 2 will test the hypothesis that if DNA-SPPs are decorated with glycosyl hydrolase enzymes (which are widely used to disrupt the biofilm matrix, specifically in the case of P. aeruginosa), they will degrade the biofilm matrix as they move through it, weakening the protection the EPS normally provides to bacteria. The success of Aim 2 will be marked by greater efficacy of a model antibiotic (ceftazidime, which has demonstrated efficacy at treating P. aeruginosa biofilms) administered topically. In Aim 3, we will load ceftazidime directly onto DNA-SPPs using a pH-sensitive motif (e.g., I-motif) that undergoes structural changes in response to pH decrease, thus releasing cargo only in acidic regions. By correlating the delivered payload to the pH distribution, we will confirm the ability of DNA-SPPs to deliver cargo preferentially in acidic regions, where hard- to-reach bacteria tend to cluster. Finally, we will assess the combinatorial benefits of the approaches in Aims 2 and 3 by using DNA-SPPs to both increase the biofilm’s permeability and to deliver antibiotics deep inside the biofilm. The major output of this study will be design criteria for DNA-based enzyme-powered SPPs to disrupt and deliver cargo in extracellular matrix (ECM) environments, which could have a major impact on the treatment of biofilms, and will lay the foundation for a customizable platform technology applicable to a wide range of ECM- mediated diseases.

项目概要/摘要细菌生物膜是大多数人类感染的罪魁祸首，导致数万人死亡和数十亿人死亡。每年的医疗费用仅通过在开放性伤口上生长而对患者造成重大伤害，众所周知，皮肤损伤、烧伤或糖尿病性溃疡等很难根除。很大程度上是因为细胞外聚合物（EPS），一种自身产生的细胞外基质，其中 EPS 在许多方面对细菌有好处，包括介导群体感应，提供营养物质，并阻断抗生素的运输和宿主免疫反应的能力。 EPS 并在最需要的地方运送抗菌货物将绕过许多这些保护措施，并可能从而对生物膜的修复产生变革性影响，于2004年首次推出人工自走式。颗粒（SPP）可以通过复杂的生物介质推动自身并将货物运送到特定位置。因此，SPP 在生物医学应用（例如生物膜修复）方面具有巨大潜力。必须克服生物相容性、跟踪和控制方面的重大挑战，才能应用于临床在这里，我们建议利用新兴的 DNA 纳米技术领域来开发脲酶动力。用于生物膜修复的基于 DNA 折纸的自推进颗粒 (DNA-SPP) 作为模型生物，我们重点关注。本研究的目标 1 将量化对经过充分研究的病原体铜绿假单胞菌的依赖性。 DNA-SPP 在局部尿素浓度和 pH 值上的运动并阐明它们的作用程度目标 2 将检验 DNA-SPP 是否用糖基修饰的假设。水解酶（广泛用于破坏生物膜基质，特别是铜绿假单胞菌），当它们穿过生物膜基质时，它们会降解生物膜基质，削弱 EPS 通常提供的保护 Aim 2 的成功将以模型抗生素（头孢他啶，它具有更强的功效）为标志。治疗铜绿假单胞菌生物膜的功效）在目标 3 中，我们将加载头孢他啶。使用 pH 敏感基序（例如 I-基序）直接连接到 DNA-SPP 上，该基序会因响应而发生结构变化通过将输送的有效负载与 pH 值相关联，从而仅在酸性区域释放货物。分布，我们将确认 DNA-SPP 优先在酸性地区运送货物的能力，在这些地区，最后，我们将评估目标 2 中方法的组合效益。 3 通过使用 DNA-SPP 来增加生物膜的渗透性并将抗生素输送到生物膜深处这项研究的主要成果将是基于 DNA 的酶驱动的 SPP 破坏的设计标准。并在细胞外基质（ECM）环境中输送货物，这可能对治疗产生重大影响生物膜，并将为适用于广泛 ECM 的可定制平台技术奠定基础介导的疾病。