Engineering Next-Generation Nanoparticles One Layer at a Time

项目摘要/摘要 癌症的治疗目前依赖于手术、放射和全身化疗。虽然这些技术 极大地改进了癌症治疗，但它们也有破坏健康组织的风险，而且消除不彻底 癌症的症状。使用纳米颗粒(NPs)作为药物输送载体可以通过以下具体方式减少这些问题 积聚在肿瘤组织中。进一步的纳米粒可以提高药物的生物利用度，扩大药物的范围 癌症治疗的潜在疗法。尽管在NP领域取得了一些成功，导致了 对于临床批准的配方，大多数依赖于被动的蓄积手段和表面 与聚乙二醇链的共轭。不幸的是，被动积累可能不会让一些人受益。 癌症类型和最近在商业产品中广泛使用的聚乙二醇酯导致了抗聚乙二醇抗体的流行 人群中有可能降低聚乙二醇基治疗药物疗效的抗体。相应地，有一个很大的 需要设计下一代具有改进性能的纳米粒子，用于癌症治疗，而不使用聚乙二醇。 一种很有前途的抗癌药物纳米粒系统是层层(LBL)纳米粒，它已经显示出巨大的前景 在癌症的临床前模型中，作为小分子、核酸或大分子的运输工具。Lbl 由一种简单的组装方法组成，包括交替吸附水中的聚合物物种到 一种可以通过静电、氢键或其他分子相互作用来调节的底物。这 这一过程允许对纳米粒子进行简单的表面修饰，这已被证明能够使癌细胞靶向和 来控制亚细胞定位。然而，人们对如何监测和控制环境缺乏了解。 拆解LBL结构以提高NP的稳定性并实现药物的精确时空控制 通过LBL-NPs交付。在F99阶段，我将探索如何以逐层方式调整层体系结构- 层(LBL)NP。在本项目中，分层和其他关键层中的溶液条件的影响 将对其特征进行调查。粒子将携带白细胞介素12，一种强大的免疫刺激 蛋白质，以评价体外和体内卵巢转移瘤的优化配方的治疗效果 癌症模型。在K00阶段，重点将从系统稳定过渡到表征 针对癌症基因治疗的细胞摄取和细胞内分解。基因治疗已经有了 在过去的几十年里，它有了许多令人兴奋的新突破，但它在癌症治疗中的应用一直受到限制 靶向性差，转染率低。我将设计一个NP配方的库，并描述它们的特征 体外和体内摄取和细胞内分解以确定可调节基因的关键NP属性 治疗效果。此外，我还将设计和优化核酸组合的新免疫疗法 通过优化的基因治疗配方进行传递的构建。

Project Summary/Abstract Cancer treatment currently relies on surgery, radiation, and systemic chemotherapy. While these techniques have greatly improved cancer therapy, they also risk damaging healthy tissue and have incomplete elimination of the cancer. The use of nanoparticles (NPs) as drug delivery vehicles may reduce these issues by specifically accumulating in tumor tissue. Further NPs can improve the bioavailability of drugs, widening the range of potential therapeutics for cancer treatment. Although there have been some successes in the NP field that led to clinically approved formulations, most have relied on passive means of accumulation and depend on surface conjugation with polyethylene glycol (PEG) chains. Unfortunately, passive accumulation may not benefit some cancer types and recent wide-spread use of PEG in commercial products has led to prevalence of anti-PEG antibodies in the population which risk reducing efficacy of PEG-based therapeutics. Accordingly, there is a great need to engineer next-generation NPs with improved properties for cancer treatment without the use of PEG. One promising NP system for cancer drug delivery is layer-by-layer (LbL) NPs which have shown great promise in preclinical models of cancer as a delivery vehicle for small molecules, nucleic acids or macromolecules. LbL consists of a simple assembly method involving the alternating adsorption of polymeric species from water onto a substrate which can be mediated by electrostatics, hydrogen-bonding or other molecular interactions. This process allows for facile surface modification of NPs which has been shown to enable cancer cell targeting and to control subcellular localization. However, there is a dearth of knowledge on how to monitor and control the disassembly of the LbL structure to improve the NP stability and enable precise spatiotemporal control of drug delivery via LbL-NPs. During the F99 phase, I will explore how to modulate the layer architecture in layer-by- layer (LbL) NPs. In this project, the effects of solution conditions during layering and other key layer characteristics will be investigated. Particles will be loaded with interleukin-12, a potent immunostimulatory protein, to evaluate treatment efficacy of optimized formulations in vitro and in an in vivo metastatic ovarian cancer model. During the K00 phase, the focus will transition from systemic stability towards characterization of cellular uptake and intracellular disassembly targeted at gene therapy for cancer treatment. Gene therapy has had many new exciting breakthroughs in the last decades, but its use in cancer treatment has been limited due to poor targeting and low transfection efficacy. I will design a library of NP formulations and characterize their uptake and intracellular disassembly in vitro and in vivo to determine key NP properties that can modulate gene therapy efficacy. Further, I will design and optimize nucleic acid combinations of new immunotherapeutic constructs to deliver via the optimized gene therapy formulations.