Engineering recombinant lubricin to combat orthopedic infection

工程重组润滑素对抗骨科感染

基本信息

批准号：
10659014
负责人：
Heidi Reesink
金额：
$ 7.85万
依托单位：
CORNELL 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：
2022-07-15 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10659014
关键词：
Acute suppurative arthritis due to bacteria Adhesions Alloys Amputation Animal Model Anti-Bacterial Agents Antimicrobial Resistance Applications Grants Arthrodesis Attenuated Award Bacteria Bacterial Adhesion Biocompatible Materials Biological Assay Biopolymers C-terminal Cell Line Clinical Codon Nucleotides Coupled Culture Media Data Debridement Deterioration Discipline Down-Regulation Engineering Enzymes Equus caballus Excision Family suidae Funding Gastric Mucin Gene Expression Genetic Glycoengineering Glycoproteins Goals Grant Image Implant In Vitro Infection MUC5AC gene Mediating Medical Device Methodology Microbial Biofilms Modeling Modification Molecular Morbidity - disease rate Mucins Multiple Bacterial Drug Resistance Musculoskeletal National Institute of Arthritis, and Musculoskeletal, and Skin Diseases Operative Surgical Procedures Organism Orthopedics Parents Patients Periprosthetic joint infection Phenotype Polysaccharides Prevalence Prevention Prevention strategy Production Proliferating Property Prosthesis Pseudomonas aeruginosa Recombinants Research Research Priority Role Sialic Acids Staphylococcus aureus Structure Surface Surface Properties Synovial Fluid Testing Therapeutic Titanium Traumatic Arthropathy United States National Institutes of Health Work antagonist antimicrobial drug clinical translation clinically relevant combat design glycosylation glycosyltransferase implant associated infection in vivo in vivo Model infection rate insight joint infection loss of function lubricin model organism mortality mucinase novel overexpression prevent sialylation transcriptome sequencing transcriptomics virulence gene

项目摘要

PROJECT SUMMARY / ABSTRACT This application seeks funding to support a NIAMS K08 recipient during her transition to research independence. The broad objective is to investigate how lubricin attenuates orthopedic biofilms and to determine whether distinct lubricin O-glycans mediate these anti-biofilm properties. The ability of many orthopedic implant-associated bacteria to form biofilms renders these organisms resistant to antimicrobial therapy. Once established, implant- associated biofilms typically require debridement, implant exchange or implant removal and can lead to severe functional deterioration necessitating arthrodesis or even amputation. Therefore, prevention and treatment of orthopedic-associated biofilms has been identified as a critical research priority in musculoskeletal infection. Recent work has identified mucin biopolymers as potent antagonists of biofilm assembly and virulence gene expression, raising the possibility that recombinant lubricin therapeutics could be an effective strategy for clinically relevant biofilms. Our preliminary data demonstrate that the recombinant lubricin developed as part of the applicant’s K08 research not only inhibits biofilm formation but also dissipates established P. aeruginosa biofilms in vitro. In the current proposal, we will investigate the hypothesis that the dual properties of surface adhesion (N- and C-terminal domains) and anti-adhesion (mucin O-glycan domains) make lubricin an ideal anti- biofilm agent. In Aim 1, we will investigate the mechanisms by which lubricin inhibits biofilm formation and induces dispersion of established biofilms in growth media, in synovial fluid, and on titanium alloy (Ti6Al4V). In Aim 2, we will determine whether distinct O-glycans mediate these anti-biofilm functions and whether lubricin functionality could be optimized through glycoengineering. As part of Aim 2, we will manipulate the expression of key glycosyltransferase enzymes to produce recombinant lubricin with altered compositions of Core-1, Core- 2 and sialylated O-glycan structures. This proposal builds upon the applicant’s K08 research, in which lubricin has been primarily investigated as a tribological agent for the treatment of post-traumatic osteoarthritis. By investigating a new, anti-biofilm application for lubricin therapy, this award will enable Dr. Reesink to branch into a new sub-discipline of orthopedic research with the potential to integrate with the applicant’s clinical veterinary expertise in small and large animal models of orthopedic infection. Dr. Reesink’s proposal is supported by an exceptional team, including experts in glycoengineering, molecular characterization of bacteria-surface interactions, and in vivo models of periprosthetic joint infection and clinical translation.

项目概要/摘要该申请寻求资金支持 NIAMS K08 接受者向独立研究过渡。广泛的目标是研究润滑素如何减弱骨科生物膜并确定是否有不同的作用润滑素 O-聚糖介导这些抗生物膜特性的能力与许多骨科植入物相关。细菌形成生物膜，使这些生物体一旦建立，就会对抗菌治疗产生耐药性。相关的生物膜通常需要清创、更换植入物或移除植入物，并可能导致严重的后果功能恶化需要关节融合术甚至截肢，因此，预防和治疗。骨科相关生物膜已被确定为肌肉骨骼感染的关键研究重点。最近的工作已确定粘蛋白生物聚合物是生物膜组装和毒力基因的有效拮抗剂表达，提高了重组润滑素疗法可能成为有效策略的可能性我们的初步数据表明，重组润滑素是作为生物膜的一部分而开发的。申请人的K08研究不仅抑制生物膜形成，还驱散已建立的铜绿假单胞菌在当前的提案中，我们将研究表面的双重特性的假设。粘附（N 端和 C 端结构域）和抗粘附（粘蛋白 O 聚糖结构域）使 lubricin 成为理想的抗粘附剂在目标 1 中，我们将研究 lubricin 抑制生物膜形成的机制。诱导已建立的生物膜分散在生长培养基、滑液和钛合金 (Ti6Al4V) 中。目标 2，我们将确定不同的 O-聚糖是否介导这些抗生物膜功能以及润滑素是否介导作为目标 2 的一部分，我们将操纵表达式关键糖基转移酶以产生具有改变的 Core-1、Core- 组成的重组润滑素 2和唾液酸化O-聚糖结构。该提案以申请人的 K08 研究为基础，其中润滑素主要作为一种通过研究一种新的抗生物膜应用来治疗创伤后骨关节炎。对于润滑素疗法，该奖项将使 Reesink 博士能够涉足骨科研究的新子学科有潜力与申请人在小型和大型动物模型方面的临床兽医专业知识相结合 Reesink 博士的建议得到了包括骨科感染专家在内的杰出团队的支持。糖工程、细菌-表面相互作用的分子表征以及细菌的体内模型假体周围关节感染和临床转化。