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 同种异体实体器官移植围手术期炎症加重急性细胞介导排斥反应 增加晚期移植物丢失，主要由慢性排斥反应引起。围手术期的两个最常见的原因 炎症是缺血再灌注损伤(IR)，在致敏患者中，预形成的供体特异性炎症 抗体，两者都在移植物上沉积抗体和补体膜攻击复合体(MACs) 内皮细胞(ECs)。MAC沉积通过诱导IL-15/IL-15的表达与排斥反应增加有关 EC表面的15Rα复合体，在那里它们可以反式呈现给宿主同种异体反应的淋巴细胞， 增强T细胞反应。在人类免疫系统小鼠模型中，这会导致更多的排斥反应 人体动脉间置移植物。我们假设，通过以下方式阻止ECs对Mac的这种特殊反应 治疗移植物而不是受者将减少早期排斥反应和晚期移植物丢失。 不断增加的全身免疫抑制。在目标1中，我们将开发和优化新的抗体靶向， 可降解的聚合物纳米颗粒(NPs)，用于选择性地将治疗性RNA输送到ECs，从而防止IL-2 15/IL-15Rα复合体。我们将评估在培养细胞和灌流细胞中的效果和持续时间。 人体血管节段。被测试的试剂包括由于从NP持续释放而将 产生持续的靶蛋白击倒，以及编码CaS酶和引导的mRNAs 可以产生永久性基因破坏或表观遗传沉默的链。内皮细胞的通路和转导 与转导分离的细胞或血管节段相比，在人体器官中的转导更具挑战性。在目标2中，我们将 进一步开发优化向肾脏和心脏内皮细胞递送抗体靶向NPs的方法 已经拒绝了使用体外常温机器的既定方法进行移植 灌流。这些实验将利用我们团队已经取得的进展，例如纤溶清除 纤维蛋白原/红细胞闭塞移植物血管以增加进入整个血管系统和 使用单体适配器改进了靶向抗体的偶联，大大增强了与内皮细胞的结合。在……里面 目的三，我们将直接检验预防IL-15反式递呈可以防止 应用我们已建立的人体动脉节段模型评估围手术期损伤的后果 人、鼠免疫系统受体的间置移植物和新的异位移植模型 体外灌流后的小鼠心脏。不管我们的IL-15假说的有效性如何， 在所有三个目标中开发的技术都可以很容易地针对其他EC目标进行调整，以实现以下任一目标 补充或代替靶向IL-15反式递呈。