Targeting Nanoparticles for Drug Delivery to Renal Graft Endothelium during Ex Vivo Normothermic Perfusion

体外常温灌注期间靶向纳米颗粒将药物递送至肾移植物内皮

• 批准号: 9164300
• 负责人: JORDAN S POBER
• 金额: $ 25.13万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-18 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9164300
• 关键词: Acute; Adaptor Signaling Protein; Address; Allografting; Amines; Antibodies; Binding; Blood; Blood Vessels; Body Temperature; Brain Death; Cell Culture Techniques; Cell Death; Cell Line; Cells; Clinic; Development; Drug Delivery Systems; Elements; Encapsulated; Endothelial Cells; Endothelium; Esters; Failure; Formulation; Frequencies; Funding; Gene Expression; Glycolic-Lactic Acid Polyester; Goals; Graft Rejection; Graft Survival; Grant; Human; Immune system; Immunosuppression; In Vitro; Individual; Inflammation; Inflammatory; Injury; Kidney; Kidney Transplantation; Link; Living Donors; Mediating; Mediator of activation protein; Metabolic; Methods; Microfluidic Microchips; Molecular Target; Organ; Organ Donor; Organ Transplantation; Outcome; Perfusion; Perioperative; Polymers; Population; Procedures; Process; Production; Pump; Recruitment Activity; Reperfusion Injury; Resources; Risk; Severities; Small Interfering RNA; Solid; Specificity; Staging; Study Section; System; TNF gene; TNFRSF1A gene; TRADD gene; Testing; Tissues; Transplantation; Tumor Necrosis Factor Receptor; United States National Institutes of Health; Universities; Work; allograft rejection; antibody conjugate; biodegradable polymer; cellular targeting; clinical application; cytokine; delayed graft function; design; immunogenicity; implantation; improved; innovation; interest; kidney allograft; knock-down; nanoparticle; neutrophil; novel; novel therapeutics; protein expression; renal ischemia; research study; response; uptake

PROJECT SUMMARY Rejection remains the major cause of allograft loss and its frequency and severity are exacerbated by peri- operative ischemia/reperfusion injury (IRI) of the graft. A mechanistic explanation of this connection is that IRI of graft endothelial cells (EC) enhances their ability to stimulate the host immune system, exacerbating rejection. Ex vivo normothermic perfusion (EVNP) is an experimental approach, pioneered for renal transplantation by our collaborators at the University of Cambridge, to reduce graft injury by improving the metabolic state of the tissue. We hypothesize that this procedure can simultaneously be used to ameliorate IRI and reduce rejection by delivery of siRNA to graft EC prior to organ implantation. We have selected tumor necrosis factor receptor 1 (TNFR1) and its key adaptor protein, TRADD, as molecular targets for siRNA knockdown in these experiments because these molecules mediate tumor necrosis factor- responses of EC that promote inflammation and cell death, important processes in renal IRI, but the goal of this R21 is limited to optimization of the effect of siRNA knockdown. siRNA delivery in the clinic is currently limited by incomplete extent and short duration of knockdown, by inability to efficiently target the cells of interest and by effects of inappropriate cellular targets. Here, we propose proof-of-principle studies to demonstrate that antibody- conjugated, degradable polymer nanoparticles (NP) can be designed to overcome these limitations. Our specific aims are: (1) to optimize formulation of multifunctional, degradable NPs using both established and novel degradable polymers for siRNA loading, siRNA sustained release and mAb conjugation; (2) to identify the best EC specific mAb or combination of mAb for binding and uptake of conjugated NP by cultured human EC under flow in vitro at Yale and in deceased donor kidneys undergoing EVNP, the latter to be performed by our collaborators at the University of Cambridge with resources independent of this R21 application; Successful completion of these aims will provide proof-of-concept for an innovative approach to reduce allograft rejection by ex vivo targeting the graft with novel drug delivery vehicles, sparing the host from the need to excessive immunosuppression.

项目摘要 排斥反应仍然是同种异体移植物丢失的主要原因，其频率和严重程度因移植物的过度使用而加剧。 移植物的手术缺血/再灌注损伤（IRI）。对这种联系的一种机械解释是， 移植物内皮细胞（EC）的增加增强了它们刺激宿主免疫系统的能力， 排斥反应离体常温灌注（EVNP）是一种实验方法，开创了肾脏移植的新方法。 我们的合作者在剑桥大学的移植，以减少移植损伤，通过改善 组织的代谢状态。我们假设，这一程序可以同时用于改善IRI 并通过在器官植入之前将siRNA递送至移植EC来减少排斥。我们选择了肿瘤 坏死因子受体1（TNFR 1）及其关键衔接蛋白TRADD作为siRNA的分子靶标 在这些实验中敲低，因为这些分子介导EC的肿瘤坏死因子-β反应 促进炎症和细胞死亡，这是肾脏IRI的重要过程，但R21的目标仅限于 siRNA敲低效果的优化。目前，临床中的siRNA递送受到不完全的限制。 由于不能有效靶向感兴趣的细胞以及由于 不合适的细胞目标在这里，我们提出了原理验证研究，以证明抗体- 可以设计共轭的、可降解的聚合物纳米颗粒（NP）来克服这些限制。我们 具体目标是：（1）使用已建立的和 用于siRNA装载、siRNA持续释放和mAb缀合的新型可降解聚合物;（2）鉴定 用于培养的人结合和摄取缀合的NP的最佳EC特异性mAb或mAb的组合 在耶鲁大学的体外流下EC和接受EVNP的死亡供体肾脏中，后者将由 我们在剑桥大学的合作者，他们的资源与R21的应用无关; 这些目标的成功实现将为减少温室气体排放的创新方法提供概念验证。 通过用新的药物递送载体离体靶向移植物，使宿主免受移植物排斥， 需要过度的免疫抑制。