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

将定制的 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

• 批准号: 10190147
• 负责人: Laura Bracaglia
• 金额: $ 10.57万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-15 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10190147
• 关键词: Address; Amines; Area; Arteries; Award; Biocompatible Materials; Biology; Biomedical Engineering; Blood Vessels; Blood capillaries; Cell Adhesion Molecules; Cell Communication; Cells; Characteristics; Charge; Chemicals; Clinical; Coupling; Development; Doctor of Philosophy; Drug Delivery Systems; Endothelial Cells; Endothelium; Environment; Extracellular Matrix; Family; Fibrosis; Formulation; Goals; Health; Human; Hydrogels; Hypoxia; Immune; Immunology; Immunosuppressive Agents; In Vitro; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injections; Injury; Injury to Kidney; Intercellular adhesion molecule 1; Kidney; Kidney Transplantation; Kinetics; Knowledge; Lead; Leukocytes; Long-Term Effects; Measures; Mediating; Mentors; Methods; Molecular; Natural regeneration; Neutrophil Infiltration; Nucleic Acids; Organ; Organ Model; Organ Transplantation; Outcome; Pathology; Penetration; Perfusion; Phase; Placenta; Polymers; Postdoctoral Fellow; Production; Reperfusion Injury; Risk; Route; Science; Signal Transduction; Site; Small Interfering RNA; Source; Structure-Activity Relationship; System; Technical Expertise; Therapeutic; Tissue Model; Tissues; Training; Transfection; Translating; Transplantation; Universities; Vascular Cell Adhesion Molecule-1; Work; base; cell injury; clinical translation; density; design; graft failure; graft function; human model; human tissue; improved; in vivo; in vivo Model; inhibitor/antagonist; injured; insight; kidney cortex; knock-down; meetings; nanoparticle; nanoparticle delivery; novel; nucleic acid delivery; prevent; regeneration potential; renal damage; resilience; response; response to injury; scaffold; siRNA delivery; small molecule; targeted treatment; therapeutic siRNA; therapy duration; three-dimensional modeling; tool; translation to humans; treatment strategy; uptake; vascular inflammation

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）引起内皮炎症和微血管稀疏，从而导致 器官移植中肾移植的不良结果。直接治疗内皮细胞（EC）可以减少 IRI对移植物健康的影响，但缺乏有效的EC靶向治疗 干预和缓解功能失调的内皮反应的各种模式。这项工作的目标是 制定一种治疗策略，解决 IRI 引起的内皮损伤的两种关键模式： 内皮细胞功能失调性炎症和毛细血管网络损伤，以特定部位和暂时的方式。 我们建议使用聚合物纳米颗粒将治疗性 siRNA 直接递送至内皮细胞 （NP），它提供了一个可定制的平台来增强细胞渗透并维持 核酸。在目标 1 中，我们将利用新型 PACE 聚合物系列来确定 NP 特性，该聚合物 使内皮细胞能够最大限度地持续使用 siRNA，以减少粘附分子的表达 炎症激活后。在目标 2 中，我们将转化这些关于结构/功能关系的知识 NP在3D人体相关模型中合理设计siRNA介导的粘附分子敲低 血管系统并评估体内移植后的长期效果。在颁奖的R00阶段， 目标 1 和 2 中确定的影响内皮-纳米颗粒相互作用的原则将应用于聚合物纳米颗粒 在水凝胶递送载体内递送至肾皮质。目标 3 将研究潜力 内皮定制的 siRNA-NP，可在 ECM 衍生的水凝胶内局部递送抗纤维化 siRNA 至 IRI- 体内肾皮质受损。 Laura Bracaglia 博士获得了生物工程博士学位，目前是该研究所的博士后研究员 耶鲁大学生物医学工程系。在迄今为止的培训中，她学习了 NP 和药物 人体组织模型中的递送方法为血管炎症提供可转化的见解。到 为了成功实现该提案的具体目标，布拉卡利亚博士确定她需要 1) 纳米颗粒开发涉及的化学和聚合物科学方面的额外培训。在 此外，拟议工作的影响将通过以下方面的培训和专业知识得到加强：2) 血管 免疫生物学，3）肾脏病理学和对损伤的反应，以及4）转化为人类免疫学。训练于 在这些领域，Bracaglia 博士组建了一支专家导师团队，他们可以提供临床观点和 技术专长。此外，她还规划了关键课程工作并为科学进步设定了里程碑 和职业目标。拟议的 K99 指导阶段培训将支持实现初始目标 这项工作的。在指导阶段制定的 NP 治疗策略以及她在 基于 ECM 的生物材料的开发，将支持该提案的最终目标 (R00)。