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.

项目概要 排斥仍然是同种异体移植失败的主要原因，其频率和严重程度因周围环境而加剧。 移植物的手术缺血/再灌注损伤（IRI）。这种联系的机械解释是 IRI 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，从而加剧了 拒绝。离体常温灌注 (EVNP) 是一种实验方法，首创用于肾病治疗 由我们在剑桥大学的合作者进行的移植，通过改善 组织的代谢状态。我们假设该程序可以同时用于改善 IRI 并通过在器官植入前将 siRNA 递送至移植物 EC 来减少排斥反应。我们选择了肿瘤 坏死因子受体 1 (TNFR1) 及其关键接头蛋白 TRADD 作为 siRNA 的分子靶标 这些实验中的敲低是因为这些分子介导 EC 的肿瘤坏死因子-α 反应 促进炎症和细胞死亡，这是肾 IRI 的重要过程，但该 R21 的目标仅限于 siRNA敲低效果的优化。 siRNA在临床上的递送目前受到不完整的限制 敲低的程度和持续时间短，由于无法有效地靶向感兴趣的细胞以及 不适当的细胞目标。在这里，我们提出原理验证研究来证明抗体- 共轭、可降解聚合物纳米颗粒 (NP) 的设计可以克服这些限制。我们的 具体目标是：（1）利用已建立的和可降解的纳米颗粒来优化多功能、可降解纳米颗粒的配方。 用于 siRNA 负载、siRNA 缓释和 mAb 缀合的新型可降解聚合物； (2) 识别 用于培养人类结合和摄取缀合 NP 的最佳 EC 特异性 mAb 或 mAb 组合 在耶鲁大学进行体外流动 EC 以及在接受 EVNP 的已故捐献肾脏中进行 EC，后者将由 我们在剑桥大学的合作者拥有独立于该 R21 应用程序的资源； 成功完成这些目标将为创新方法提供概念验证，以减少 通过用新型药物递送载体体外靶向移植物来进行同种异体移植排斥，从而使宿主免受 需要过度的免疫抑制。