Ex Vivo Nanoparticle Drug Delivery Targeted to Human Allograft Endothelium

针对人同种异体移植物内皮的体外纳米颗粒药物输送

• 批准号: 10783379
• 负责人: JORDAN S POBER
• 金额: $ 41.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-06 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10783379
• 关键词: Acute; Adaptive Immune System; Allogenic; Allografting; Antibodies; Aorta; Arteries; Binding; Blood Vessels; Cell Line; Cell Separation; Cell surface; Cells; Chronic; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Complement; Complement Membrane Attack Complex; Complex; Coupling; Cultured Cells; Cytolysis; Data; Deposition; Development; Drug Carriers; Endothelial Cells; Endothelium; Enzymes; Epigenetic Process; Erythrocytes; Esters; Fibrinogen; Fluorescent Dyes; Formulation; Frequencies; Future; Genes; Genetic; Graft Survival; Heart; Heart Transplantation; Heterotopic Transplantation; Human; Immune response; Immune system; Immunodeficient Mouse; Immunosuppression; Implant; In Vitro; Inflammation; Injury; Interleukin-15; Kidney; Kinetics; Link; Liver; Lymphocyte; Mediating; Membrane; Messenger RNA; Modeling; Mus; Organ; Organ Donor; Organ Transplantation; Pathologic; Patients; Perfusion; Perioperative; Phagocytes; Pharmaceutical Preparations; Play; Polyamines; Polymers; Postoperative Period; Prevention; Procedures; Proteins; Pump; RNA; RNA delivery; Regional Perfusion; Reperfusion Injury; Risk; Role; Sampling; Severities; Signal Transduction; Small Interfering RNA; Solid; Spleen; Surface; T cell response; T memory cell; T-Lymphocyte; Technology; Testing; Therapeutic; Time; Tissues; Transplantation; Transplantation Surgery; antibody conjugate; biodegradable polymer; donor-specific antibody; efficacy evaluation; epigenetic silencing; ex vivo perfusion; experimental study; heart allograft; human model; immunogenicity; improved; kidney allograft; kidney cell; knock-down; mRNA delivery; mouse model; nanoparticle; nanoparticle delivery; nanoparticle drug; nanopolymer; new technology; novel; pre-clinical; prevent; response; targeted delivery; therapeutic RNA; therapeutic nanoparticles

7. Project Summary/Abstract Peri-transplantation inflammation of solid organ allografts exacerbates acute cell-mediated rejection and increases late graft loss, primarily caused by chronic rejection. The two most common causes of perioperative inflammation are ischemia reperfusion injury (IR) and, in sensitized patients, pre-formed donor specific antibodies, both of which deposit antibody and complement membrane attack complexes (MACs) on graft endothelial cells (ECs). MAC deposition is linked to increased rejection by inducing expression of IL-15/IL- 15Rα complexes on the EC surface where they can be trans-presented to host alloreactive lymphocytes, intensifying T cell responses. In a human immune system mouse model, this results in increased rejection of human arterial interposition grafts. We hypothesize that preventing this specific response of ECs to MACs by treating the graft rather than the recipient will reduce early rejection episodes and late term graft loss without increasing systemic immunosuppression. In aim 1, we will develop and optimize new antibody-targeted, degradable polymer nanoparticles (NPs) for delivery of therapeutic RNA selectively to ECs that can prevent IL- 15/IL-15Rα complexes. We will evaluate efficacy and duration of effects in both cultured cells and perfused human vessel segments. Agents to be tested include siRNAs that, due to sustained release from the NP, will produce a sustained knockdown of target proteins, as well as mRNAs encoding Cas enzymes and guide strands that can produce permanent gene disruption or epigenetic silencing. Access and transduction of ECs in human organs is more challenging than transduction of isolated cells or vessel segments. In Aim 2, we will further develop approaches to optimize delivery of the antibody-targeted NPs to ECs of kidneys and hearts that have been declined for transplantation using established approaches of ex vivo normothermic machine perfusion. These experiments will exploit advances already made by our team, such as fibrinolytic clearing of fibrinogen/erythrocyte occlusions of graft vasculature to increase access to the whole vasculature and improved coupling of targeting antibodies using monobody adapters that greatly enhance binding to ECs. In aim three, we will directly test the hypothesis that prevention of IL-15 trans-presentation can prevent the consequences of peri-operative injuries using both our well established model of human artery segment interposition grafts in human immune system mouse recipients and in a new model of heterotopic transplants of mouse hearts following ex vivo perfusion. Regardless of the validity of our IL-15 hypothesis, the technologies developed in all three aims can be readily adapted for use against other EC targets to either complement or in lieu of targeting IL-15 trans-presentation.

7。项目摘要/摘要 固体器官同种异体移植物的移植周期炎症加剧了急性细胞介导的排斥和 增加谷物损失，主要是由慢性排斥反应引起的。定期的两个最常见原因 炎症是缺血再灌注损伤（IR），在敏感的患者中，预先形成的供体特异 抗体，它们都沉积抗体和补体膜攻击复合物（MAC） 内皮细胞（EC）。 MAC的沉积与诱导IL-15/IL-的表达的排斥反应有关 EC表面上的15Rα络合物，可以将它们反式传播以寄主同种异体淋巴细胞， 加强T细胞反应。在人类免疫系统鼠标模型中，这导致对 人动脉介入移植物。我们假设通过通过 治疗移植物而不是接受者将减少早期拒绝发作和后期移植物损失而无需 增加系统性免疫抑制。在AIM 1中，我们将开发和优化以新的抗体为目标， 可降解的聚合物纳米颗粒（NP）选择热RNA递送到可以防止IL-的EC 15/IL-15Rα复合物。我们将评估培养细胞和灌注的有效性和效果持续时间 人体船只段。要测试的代理包括SIRNA，由于NP持续释放，将 产生了靶蛋白的持续敲低，以及编码CAS酶的mRNA和指导 可以产生永久基因破坏或表观遗传沉默的链。 ECS的访问和翻译 在人体器官中，比分离的细胞或容器段的转移更具挑战性。在AIM 2中，我们将 进一步的开发方法，以优化将抗体靶向的NP传递给儿童和心脏的EC 使用既定的近常热机的既定方法被拒绝移植 灌注。这些实验将利用我们的团队已经取得的进步，例如 移植脉管系统的纤维蛋白原/红细胞闭塞，以增加进入整个脉管系统的机会 使用单体适配器改善了靶向抗体的耦合，从而增强了与EC的结合。 目的三，我们将直接检验以下假设：预防IL-15反式呈现可以防止 使用我们建立的人类动脉段模型的围手术损伤的后果 人类免疫系统的介入移植物和新型异位移植模型中的移植物 在体内灌注后的小鼠心脏的心脏。无论我们的IL-15假设的有效性如何 在所有三个目标中开发的技术都可以很容易地适应其他EC目标来使用 补充或代替靶向IL-15反式出现。