Multimodal nanoparticles (gold in polylactide) and their effect on three-dimensional cell cultures as organ models

多峰纳米颗粒（聚丙交酯中的金）及其对作为器官模型的三维细胞培养物的影响

• 批准号: 417844584
• 负责人: Professor Dr. Matthias Epple
• 金额: --
• 依托单位: Lehrstuhl für Infektionsimmunologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/417844584?language=en
• 关键词: Multimodal; nanoparticles; gold; polylactide; their

We will synthesise multimodal nanoparticles that can be tracked in the cell and detected in the endo-lysosome by specific fluorescence labels. This will be achieved by covalently binding fluorescently labelled biomolecules to small gold nanoparticles (diameter approx. 10 nm) where their fluorescence is quenched. These will be incorporated into polylactide-co-glycolide (PLGA) particles (estimation: approximately 10 gold nanoparticles within a single PLGA particle with a diameter of around 100 nm). The PLGA envelope will also be covalently functionalised with fluorescent dyes. The cellular uptake of the multimodal nanoparticles will be studied quantitatively using 3D cell culture models (crypt organoids and tumour spheroids) by in situ confocal laser scanning microscopy. We will focus on the pathway of the nanoparticles inside the complex cell network of the 3D cell culture or organoids and investigate the impact of the multimodal nanoparticles and biomolecules. Biomolecules for gene silencing (siRNA) will be conjugated to the gold nanoparticles to achieve a biological effect. First, the gene silencing of fluorescent eGFP as model protein will be investigated, followed by the gene silencing of important inflammatory disease mediators, such as TNF-α, KC, IP 10, and the apoptosis-inhibiting survivin as an additional protein model. The nanoparticle-mediated gene silencing of survivin will prove and extend our concept to another therapeutically relevant target.In summary, we aim for a better understanding of the biological effects of functional nanoparticles, covering inter alia the questions of dose, pathway, and fate. Three-dimensional cell culture models shall serve as bridge between "classical" two-dimensional cell cultures and animal models. We also want to achieve a quantitative validation of two-dimensional and three-dimensional cell culture models with respect to biologically active nanoparticles.

我们将合成多模式纳米颗粒，这些纳米颗粒可以在细胞内追踪，并通过特定的荧光标记在内切溶酶体中检测到。这将通过共价结合荧光标记的生物分子到小的金纳米颗粒(直径约。10 nm)，其中它们的荧光被猝灭。这些将被加入到聚乳酸-乙交酯(PLGA)颗粒中(估计：在直径约100纳米的单个PLGA颗粒中约有10个金纳米颗粒)。PLGA包膜还将与荧光染料共价官能化。多模式纳米粒子的细胞摄取将使用3D细胞培养模型(隐窝器官和肿瘤球体)通过原位共聚焦激光扫描显微镜进行定量研究。我们将重点研究纳米颗粒在3D细胞培养或有机物的复杂细胞网络中的路径，并研究多峰纳米颗粒和生物分子的影响。用于基因沉默的生物分子(SiRNA)将被偶联到金纳米颗粒上，以实现生物效应。首先，将研究作为模型蛋白的荧光绿色荧光蛋白的基因沉默，其次是重要的炎症性疾病介质，如肿瘤坏死因子-α、KC、IP10的基因沉默，以及作为附加蛋白模型的抑制凋亡的Survivin。纳米颗粒介导的Survivin基因沉默将证明我们的概念，并将其扩展到另一个具有治疗意义的靶点。综上所述，我们的目标是更好地了解功能纳米颗粒的生物学效应，包括剂量、途径和命运等问题。三维细胞培养模型将成为“经典的”二维细胞培养和动物模型之间的桥梁。我们还希望实现与生物活性纳米颗粒相关的二维和三维细胞培养模型的定量验证。