Nanostructures and imaging tools for detection of cancer cell surface markers

用于检测癌细胞表面标志物的纳米结构和成像工具

• 批准号: RGPIN-2014-05199
• 负责人: Zou, Shan
• 金额: $ 2.19万
• 依托单位: Carleton University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=669979
• 关键词: Nanostructures; imaging; tools; detection; cancer

We will create nanostructures and polymer materials, explore self-assembly, to improve surface binding specificity and enable high resolution imaging of cell membranes and their constituents. We aim to employ chemically and mechanically controlled surface functionalization for probing fundamental problems in biophysical chemistry of cellular processes, establishing a novel platform for bio-imaging, protein adhesion and cell mechanics. Controlled surface chemistry will introduce molecular recognition and specificity. Integration with cell mechanics probed by atomic force microscopy (AFM) will bring new insights into cellular processes such as force transduction across cell membranes at the 50 nm length scale, which is not possible to probe for heterogeneous systems using existing approaches. In particular, our newly developed self-assembly and seeding growth method is suitable for low-cost mass-manufacturing of ordered nanoparticle arrays, and the proposed hybrid nanostructure using similar approach will provide high sensitivity for plasmonic enhanced subwavelength imaging and sensing application. To integrate this novel nanoparticle based platform with the understanding on mechanical properties of cancer cells at the molecular resolution is the long term goal of our research, which will establish the structure-property-function relationship that enable the design of new materials and devices for cancer diagnosis and prognosis, and fulfill the requirement of new metrologies incisive on the nanoscale. *Three research areas with specific projects are proposed in this program, namely, *Area 1. Mechanical detection of the acute promyelocytic leukemia (APL) cell membranes: *(1a). Fabrication of microwells for non-adherent APL cells; *(1b). Elasticity measurements of drug treated and non-treated APL cell line (NB4) cells;*(1c). Therapeutic treatments and viability evaluations on drug resistant cells;*(1d). Mechanical detection on APL patient samples.*Area 2. Single molecule force spectroscopy (SMFS) study of the retinoic acid (RA) receptor and retinoic acid binding interactions:*(2a). Binding interaction between all trans retinoic acid (ATRA) and RA receptors;*(2b). Unbinding forces of the individual complexes of the drug-RA receptor and the ligand-RA receptor.*Area 3. Nanostructure platform for antibody, membrane protein and cell surface marker detection:*(3a). Hybrid nanoparticle (NP) arrays for anti-surface marker (CD38) detection; *(3b). Local surface plasmon resonance coupled fluorescence for quantification of the surface markers (CD38 and CD56).**The outlined program will include the training of two graduate and five undergraduate students. The current undergraduate and Ph.D students will also benefit from the team working environment and scientific discussions proposed in this program. All students will be trained with expertise in surface chemistry, self-assembly, and advanced atomic force microscopy and integrated fluorescence microscopy for membrane characterizations and sensing applications. **The full potential of using nanostructures and novel materials in biophysical chemistry and cell biology is predicted to be attainable when combined with advanced microscopy and spectroscopy techniques. The proposed multi-modal imaging and single molecule mechanical detection approach will allow cellular structures to be identified by fluorescence, imaged at high resolution using AFM, and mechanically probed by single molecule force spectroscopy. Complementary to conventional ensemble measurements, surface chemistry and scanning probe microscopy based techniques will provide new pioneering insights in understanding the molecular principles of cancer and nanomedicine.

我们将创造纳米结构和聚合物材料，探索自组装，以提高表面结合特异性，并实现细胞膜及其成分的高分辨率成像。我们的目标是利用化学和机械控制的表面功能化来探索细胞过程中生物物理化学的基本问题，建立生物成像，蛋白质粘附和细胞力学的新平台。受控表面化学将引入分子识别和特异性。原子力显微镜（AFM）与细胞力学的结合将为细胞过程带来新的见解，例如在50纳米长度尺度上跨细胞膜的力转导，这是现有方法无法探测异质系统的。特别是，我们新开发的自组装和播种生长方法适用于有序纳米颗粒阵列的低成本批量制造，并且采用类似方法提出的混合纳米结构将为等离子体增强亚波长成像和传感应用提供高灵敏度。将这种基于纳米粒子的新型平台与在分子分辨率上对癌细胞力学特性的理解相结合是我们研究的长期目标，它将建立结构-性能-功能关系，为癌症诊断和预后的新材料和设备的设计提供可能，并满足新计量学在纳米尺度上的需求。*本项目提出三个研究领域，具体项目为*Area 1。急性早幼粒细胞白血病（APL）细胞膜的机械检测：*(1a)。非贴壁APL细胞微孔的制备* (1 b)。药物处理和未处理APL细胞系（NB4）细胞的弹性测量；*(1c)。耐药细胞的治疗和活力评估；*(1d)。APL患者标本的力学检测。*区域2。维甲酸（RA）受体与维甲酸结合相互作用的单分子力谱（SMFS）研究：*(2a)。所有反式维甲酸（ATRA）和RA受体之间的结合相互作用；*(2b)。药物类风湿性关节炎受体和配体类风湿性关节炎受体的单个复合物的解结合力。* 3。抗体、膜蛋白和细胞表面标记物检测的纳米结构平台：*(3a)。用于抗表面标记物（CD38）检测的杂化纳米颗粒（NP）阵列* (3 b)。局部表面等离子体共振耦合荧光定量表面标记（CD38和CD56）。**概述的计划将包括培训两名研究生和五名本科生。目前的本科生和博士生也将受益于这个项目提出的团队工作环境和科学讨论。所有学生都将接受表面化学、自组装、先进原子力显微镜和集成荧光显微镜等方面的专业知识培训，用于膜表征和传感应用。**与先进的显微镜和光谱学技术相结合，纳米结构和新材料在生物物理化学和细胞生物学方面的全部潜力有望实现。提出的多模态成像和单分子机械检测方法将允许通过荧光识别细胞结构，使用AFM进行高分辨率成像，并通过单分子力谱进行机械探测。作为传统集合测量的补充，基于表面化学和扫描探针显微镜的技术将为理解癌症和纳米医学的分子原理提供新的开创性见解。