Engineering Next-Generation Nanoparticles One Layer at a Time

项目总结/摘要 癌症治疗目前依赖于手术、放疗和全身化疗。虽然这些技术 大大改善了癌症治疗，但它们也有损害健康组织的风险， 癌症使用纳米颗粒（NPs）作为药物递送载体可以通过特异性地降低这些问题。 在肿瘤组织中积聚。进一步的NP可以提高药物的生物利用度，拓宽药物的应用范围。 癌症治疗的潜在疗法。尽管在NP领域取得了一些成功， 到临床批准的制剂，大多数依赖于被动积累手段，并依赖于表面 与聚乙二醇（PEG）链缀合。不幸的是，被动积累可能不会使一些人受益。 癌症类型和最近PEG在商业产品中的广泛使用导致抗PEG抗体的流行。 这些抗体有降低基于PEG的治疗剂的功效的风险。因此，有一个伟大的 需要设计下一代纳米粒子，使其具有改善的癌症治疗性能，而无需使用PEG。 用于癌症药物递送的一种有前途的NP系统是已经显示出巨大前景的逐层（LbL）NP 在癌症的临床前模型中作为小分子、核酸或大分子的递送载体。LBL 包括一种简单的组装方法，该方法涉及将聚合物物质从水中交替吸附到 可以通过静电、氢键或其它分子相互作用介导的基质。这 该方法允许对NP进行容易的表面修饰，这已经显示出能够靶向癌细胞， 来控制亚细胞定位然而，缺乏关于如何监测和控制 LbL结构的拆卸以改善NP稳定性并能够精确地时空控制药物 通过LbL-NP递送。在F99阶段，我将探索如何逐层调整层架构， 层（LbL）NP。在本工程中，分层和其他关键层的溶解条件的影响， 特性将被研究。颗粒将装载白细胞介素-12，一种有效的免疫刺激剂， 蛋白质，以评估优化制剂在体外和体内转移性卵巢癌中的治疗功效。 癌症模型在K 00阶段，重点将从系统稳定性过渡到表征 细胞摄取和细胞内分解，靶向用于癌症治疗的基因疗法。基因治疗已 在过去的几十年里，它有许多令人兴奋的新突破，但由于其在癌症治疗中的应用有限， 靶向性差和转染效率低。我将设计一个NP配方库，并描述其 在体外和体内的摄取和细胞内分解，以确定可以调节基因的关键NP性质， 治疗效果此外，我将设计和优化新的免疫抑制剂的核酸组合， 构建体，以通过优化的基因治疗制剂递送。

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.