Tailored siRNA delivery to human endothelium to inhibit and reverse inflammatory damage following ischemia reperfusion injury in the kidney

定制的 siRNA 递送至人内皮细胞以抑制和逆转肾脏缺血再灌注损伤后的炎症损伤

基本信息

批准号：
10686146
负责人：
Laura Bracaglia
金额：
$ 24.78万
依托单位：
VILLANOVA 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：
2022-09-01 至 2025-08-31
项目状态：
未结题

项目摘要

Abstract Ischemia reperfusion injury (IRI) causes endothelial inflammation and microvascular rarefaction that leads to adverse kidney graft outcomes in organ transplant. Direct treatment of endothelial cells (EC) can reduce the impact of IRI on the health of the graft, but there is a lack of EC targeted therapies that can effectively intervene and alleviate the various modes of dysfunctional endothelial response. The goal of this work is to develop a therapeutic strategy that addresses the two key modes of endothelial damage in response to IRI: dysfunctional inflammation in ECs and damage to capillary networks, in a site-specific and temporary manner. We propose that therapeutic siRNA can be delivered directly to endothelial cells using polymeric nanoparticles (NPs), which provide a customizable platform to enhance the cell penetration and to sustain the delivery of nucleic acids. In Aim 1, we will determine the NP characteristics utilizing a novel family of PACE polymers that enable maximum and sustained siRNA to endothelial cells in order to reduce adhesion molecule expression upon inflammatory activation. In Aim 2, we will translate this knowledge of structure/function relationship of the NP to rationally design siRNA-mediated knockdown of adhesion molecules in relevant models of 3D human vasculature and evaluate the long-term effect after transplantation in vivo. In the R00 phase of the award, the principles determined in Aim 1 and 2 to impact endothelial-NP interaction will be applied to polymer NPs delivered within a hydrogel delivery vehicle to the renal cortex. Aim 3 will investigate the potential of endothelial-tailored siRNA-NPs to locally deliver anti-fibrotic siRNAs within an ECM-derived hydrogel to IRI- damaged renal cortex in vivo. Dr. Laura Bracaglia has earned her PhD in Bioengineering and is currently a postdoctoral fellow in the Department of Biomedical Engineering at Yale University. In her training so far, she has studied NP and drug delivery methods in human tissue models that provide translatable insights into vascular inflammation. To successfully accomplish the specific aims of this proposal, Dr. Bracaglia has identified that she will need additional training in the 1) chemical and polymer science aspects involved in the development of NPs. In addition, the impact of the proposed work would be enhanced with training and expertise in 2) vascular immune biology, 3) renal pathology and response to injury, and 4) translation to human immunology. To train in these areas, Dr. Bracaglia has assembled a team of expert mentors who can provide clinical perspective and technical expertise. In addition, she has planned key course work and set milestones for progress in scientific and professional goals. This proposed training in the K99 mentored phase will support meeting the initial goals of this work. NP treatment strategies developed during the mentored phase, together with her expertise in the development of ECM-based biomaterials, will support the final aim of this proposal (R00).

抽象的缺血再灌注损伤（IRI）会导致内皮炎症和微血管罕见，导致器官移植中的不良肾移植结果。直接治疗内皮细胞（EC）可以减少 IRI对移植物健康的影响，但是缺乏EC靶向疗法可以有效地干预和减轻内皮功能失调的各种模式。这项工作的目的是制定一种治疗策略，该策略解决了对IRI的响应： ECS的功能失调的炎症和毛细管网络的损害，以特定地点和临时的方式。我们建议可以使用聚合物纳米颗粒直接将治疗性siRNA直接传递到内皮细胞（NPS），它提供了一个可自定义的平台，以增强细胞渗透并维持交付核酸。在AIM 1中，我们将利用新型的速度聚合物系列来确定NP特征启用最大和持续的siRNA到内皮细胞以降低粘附分子表达炎症激活后。在AIM 2中，我们将翻译有关结构/功能关系的知识 NP至合理设计siRNA介导的粘附分子的敲低3D人类相关模型中的粘附分子脉管系统和体内移植后的长期效应。在奖励的R00阶段，在AIM 1和2中确定的原理以影响内皮NP相互作用，将应用于聚合物NP 在水凝胶输送车中递送到肾皮层。 AIM 3将调查潜力内皮量身定制的siRNA-NP，以在ECM衍生的水凝胶中局部提供抗纤维化siRNA 体内受损的肾皮质。劳拉·布拉卡利亚（Laura Bracaglia）博士在生物工程中获得了博士学位，目前是该博士学位耶鲁大学生物医学工程系。到目前为止，她在培训中学习了NP和毒品人体组织模型中的递送方法，可翻译成血管炎症的见解。到 Bracaglia博士成功地完成了该提案的具体目标，她确定她将需要在1）化学和聚合物科学方面进行的其他培训涉及NP的发展。在此外，通过培训和专业知识2）血管的培训和专业知识将增强所提议的工作的影响免疫生物学，3）肾脏病理和对损伤的反应，以及4）转化为人类免疫学。训练在这些领域，Bracaglia博士组建了一个专家导师团队，他们可以提供临床观点和技术专长。此外，她还计划了关键课程工作，并为科学进步设定了里程碑和专业目标。在K99指导阶段的拟议培训将支持实现最初的目标这项工作。 NP治疗策略在指导阶段制定了，以及她的专业知识基于ECM的生物材料的开发将支持该提案的最终目标（R00）。