Endocytosis of emerging nanomedicines: Understanding the influence of shape of 3D nanoprinted soft matter materials

职权范围：这个“基础生物科学”项目涉及BBSRC的战略研究重点3--“生物科学促进健康”，因为它旨在发展基础生物科学，以支持新出现的纳米药物的配方和有效性，这些药物将有助于维护和促进健康。问题：尺寸、形状和表面化学的相互依赖如何影响纳米材料的生物分布、细胞内化和细胞内运输，人们知之甚少。随着制造纳米药物的尖端技术的增加，如3D纳米打印，可以产生特定几何形状和变形能力的单分散产品，人们对形状、图案和变形性对细胞摄取和生物分布的影响的洞察力正在显现。表面的图案也可以影响蛋白质的吸附，从而影响细胞的反应。对比：随着2015年监管部门批准的第一种3D打印药物，Precendence被设定为在制药行业使用3D打印，人们对探索纳米级功能(即3D纳米打印)的兴趣正在迅速增加。3D打印的吸引人的优势之一是它的多功能性和多样性，可以在卫生部门开发个性化的药物。医疗保健中的3D纳米打印将能力和潜力扩展到纳米药物的RELM中。纳米药物可以使用多种途径进入细胞，这一点目前还不够了解。纳米药物有多种内吞机制，包括通过依赖于分子筛和不依赖于分子筛的途径吞噬和吞噬胞饮。PROJECT目标：该项目将探索3D纳米印刷软物质的内吞过程，并调查这些纳米材料对细胞内化和细胞内运输的形状影响。PHD工作计划：要使用的关键资源：-尼康成像中心：2012年在伦敦国王学院建立了世界尼康成像中心。作为世界上仅有的8个尼康成像中心之一，也是英国唯一的一个，GB 450万个中心拥有10台不同的尖端研究显微镜。该项目将使用尖端的研究显微技术，如尼康成像中心提供的TI2活细胞成像系统，对动植物细胞系的内吞动力学进行成像。-伦敦纳米技术中心(LCN)：LCN是伦敦的一个物理和生物医学纳米技术的多学科研究中心。伦敦国王学院最近加入了LCN，Raimi-Abraham博士正在与Richard Thorogate博士(生物纳米实验室)在软质纳米材料的定量纳米机械表征领域进行合作。该项目将探索利用高速原子力显微镜(HS-AFM)与共聚焦激光扫描单元相结合来评估内吞过程中质膜和蛋白质组装的形态变化。YEAR One(KCL)：-培训与项目相关的技术，如3D纳米打印、细胞培养、高分辨率电子显微镜(用于细胞成像及其与3D纳米打印软物质材料的相互作用)、腔内显微镜、第二年(KCL和巴斯夫)：-构建各种形状的3D纳米特征-表征表面属性，如粗糙度和了解与待研究的细胞的潜在相互作用。第三年(KCL)：-分析纳米颗粒对活的动物(如Caco-2细胞系或TR146细胞系)和植物细胞(如烟草BY-2细胞)生物学功能的影响。第四年(KCL和巴斯夫)：-反馈生物学研究的结果，以纳米尺度上的细胞与材料相互作用为基础重新定义和产生新的知识。

REMIT: This "Basic Bioscience" project addresses BBSRC's strategic research priority 3, "Bioscience for Health" as it aims to develop basic bioscience underpinning the formulation and effectiveness of emerging nanomedicines that will benefit the maintenance and promotion of health.THE PROBLEM:Little is known how the interdependency of size, shape, and surface chemistry can influence the biodistribution, cellular internalization, and intracellular trafficking of nanomaterials. Insights into the effect of shape, patterning, and deformability on cellular uptake and biodistribution are emerging with the increase in sophisticated techniques to manufacture nanomedicines such as 3D nanoprinting that can generate monodisperse products of specific geometry and deformability. Patterning on surfaces can also influence protein adsorption and consequently cellular responses.CONTEXT:With the first regulatory approved 3D printed medicine in 2015, the precendence was set for the use of 3D printing in the pharmaceutical industry and interest in exploring capabilities in the nanoscale (I.e. 3D nanoprinting) is rapidly increasing. One of the attractive advantages of 3D printing its versatility and diversity which can be exploited in the health sector to develop personalised medicines. 3D nanoprinting in healthcare extends capabilities and potential into the relm of nanomedicines.Nanomedicines can employ multiple pathways for cellular entry, which are currently insufficiently understood.Various mechanisms of endocytosis are available to nanomedicines including phagocytosis and pinocytosis through clathrin-dependent and clathrin-independent pathways.PROJECT AIMS:This project will explore to process of endocytosis of 3D nanoprinted soft matter materials and investigate the shape effect of these nanomaterials on cellular internalization, and intracellular trafficking.PhD WORK PROGRAMME:Key resources to be used:- Nikon Imaging Centre: The world renowed Nikon Imaging Centre was established in 2012 at King's College London. As one of only 8 Nikon Imaging Centres in the world and the only one in the UK, the £4.5 million centre houses ten different cutting-edge research microscopes. This project will involve imaging the dynamics of endocytosis in animal and plant cell lines using cutting edge research microscopic techniques such as the Ti2 live cell imaging systems available at the Nikon Imaging centre.- London Centre for Nanotechnology (LCN): LCN is a multidisciplinary research centre in physical and biomedical nanotechnology in London. King's College London recently joined the LCN and Dr Raimi- Abraham has an ongoing collaboration with Dr Richard Thorogate (BioNano Laboratory) in the area of quantitative nanomechanical characterisation of soft matter nanomaterials. The project will explore the assessment of morphological changes of plasma membrane and protein assembly during endocytosis utilizing high-speed atomic force microscopy (HS-AFM) combined with concofocal laser scanning unit.YEAR ONE (KCL):- Student training in project relevant techniques such as 3D nanoprinting, cell culture, high resolution electron microscopy (for imaging of cells and their interaction with 3D nanoprinted soft matter materials), intra-vtiral microscopy, atomic force microscpy and confocal microscopy).ii) Fabricate templates for 3D nanoprinting.YEAR TWO (KCL & BASF):- Formulation of 3D nanofeatures with various shapes- Characterisation of surface properties such as roughness and understanding of potential interaction with the cells for uptake.YEAR THREE (KCL):- Analyze the effects of nanopatterns on the biological function of living animal (such as Caco-2 cell line or TR146 cell line) and plant cells (such as Tobacco BY-2 cells)YEAR FOUR (KCL & BASF):- Feedback the results of biological studies to redefine and generate new knowledge based on cell- material interactions at nanometre scales.