Engineered nanoparticles to rescue complement dysfunction and vascular disease during diabetes

项目摘要/摘要 糖尿病期间的血管损伤和疾病会导致终末器官功能障碍(肾衰竭、失明、 截肢)和心血管疾病，大大降低了患者的生活质量和预期寿命。这个 拟议的工作旨在开发新的疗法来预防和遏制糖尿病血管疾病，方法是 设计纳米颗粒，使蛋白质治疗性药物能够有效传递。血管损伤的主要原因 是蛋白质糖基化，即葡萄糖与蛋白质的共价结合，导致晚期糖基化终末- 产品和蛋白质失活。尤其是膜攻击的关键调节蛋白CD59 复合体(MAC)是普遍表达的，人类CD59对糖基化失活是唯一敏感的 在糖尿病期间。人类和实验研究强调了CD59糖基化- 组织中的失活、补体失调和MAC沉积。因此，外源性的交付， 糖基化抵抗的CD59可以挽救补体调节并减轻补体介导的血管 糖尿病中的损伤，但由于这种蛋白的快速清除和非特异性血清结合，具有挑战性。 此外，提供战略需要实现广泛的细胞亲和性(红细胞、内皮细胞和 上皮细胞)和高递送效率。这项工作的目标是设计纳米颗粒(NPs)以使 利用脂基纳米粒改善蛋白质的体内高效静脉给药 药物动力学。我们假设，CD59纳米颗粒的输送将减轻心血管疾病和 CD59缺乏的糖尿病小鼠的内皮功能障碍。在目标1中，NPs将针对以下方面进行优化 生物相容性、蛋白质负载量、最小非特异性摄取和体外释放。优化后的NP将被 在Aim 2中评价预防动脉粥样硬化、内皮细胞的生物分布和治疗效果 糖尿病，CD59缺陷动物的功能障碍和组织补体沉积。成功完成 这项工作可以验证CD59作为阻止和预防糖尿病血管并发症的临床策略的有效性。 申请者加里·刘的目标是领导一个实验室，担任首席研究员。他未来的研究目标之一是 开发治疗糖尿病微血管并发症的新疗法。该博士后奖学金包括(1) 糖尿病及其并发症、内分泌学和生物信息学的教学培训；(2)科学培训 糖尿病动物模型和量化心血管疾病和内皮功能障碍的方法，以及 (3)在负责任地进行研究方面的专业发展和培训，为职业生涯做准备 首席调查员。培训将在位于罗伯特·兰格博士实验室的麻省理工学院科赫研究所进行，这将 提供执行拟议工作所需的材料、设备和智力专业知识。

PROJECT SUMMARY/ABSTRACT Vascular injury and disease during diabetes lead to end-organ dysfunction (kidney failure, blindness, amputation) and cardiovascular disease that profoundly reduces patient quality of life and life expectancy. The proposed work seeks to develop new therapeutics to prevent and arrest diabetic vascular disease, by engineering nanoparticles to enable efficient delivery of a protein therapeutic. A major cause of vascular injury is protein glycation, the covalent attachment of glucose to proteins that leads to advanced glycation end- products and protein inactivation. In particular, the protein CD59, the key regulator of the membrane attack complex (MAC), is ubiquitously expressed, and human CD59 is uniquely susceptible to glycation-inactivation during diabetes. Human and experimental studies underscore the connection between CD59 glycation- inactivation, complement dysregulation, and MAC deposition in tissues. Therefore, delivery of exogenous, glycation-resistant CD59 could rescue complement regulation and mitigate complement-mediated vascular injury in diabetes, but is challenging due to rapid clearance and non-specific serum-binding of this protein. Moreover, a delivery strategy would need to achieve broad cell tropism (red blood cells, endothelial and epithelial cells) and high delivery efficiency. The goal of this work is to engineer nanoparticles (NPs) to enable efficient intravenous delivery of mouse CD59 in vivo using lipid-based NPs to improve protein pharmacokinetics. We hypothesize that nanoparticle delivery of CD59 will mitigate cardiovascular disease and endothelial dysfunction in diabetic mice with CD59 deficiency. In Aim 1, NPs will be optimized for biocompatibility, protein loading, minimal non-specific uptake, and delivery in vitro. Optimized NPs will then be evaluated in Aim 2 for biodistribution and therapeutic efficacy in preventing atherosclerosis, endothelial dysfunction, and tissue complement deposition in diabetic, CD59-deficient animals. Successful completion of this work could validate CD59 delivery as a clinical strategy to halt and prevent diabetic vascular complications. The applicant, Gary Liu, aims to lead a lab as a principal investigator. One of his future research aims is to develop new therapies for diabetic microvascular complications. This postdoctoral fellowship includes (1) didactic training in diabetes, its complications, endocrinology, and bioinformatics; (2) scientific training in diabetic animal models and methods for quantifying cardiovascular disease and endothelial dysfunction, and (3) professional development and training in the responsible conduct of research in preparation for a career as a principal investigator. Training will occur at the Koch Institute of MIT in Dr. Robert Langer’s lab, which will provide the materials, equipment, and intellectual expertise necessary to carry out the proposed work.