Nanostructures and imaging tools for detection of cancer cell surface markers

我们将创建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。我们的目标是采用化学和机械控制的表面功能化来探测细胞过程的生物物理化学中的基本问题，建立一个新的生物成像，蛋白质粘附和细胞力学的平台。受控的表面化学将引入分子识别和特异性。与原子力显微镜（AFM）探测的细胞力学的集成将带来新的见解，如在50 nm的长度尺度，这是不可能探测异质系统使用现有的方法在细胞膜上的力转导的细胞过程。特别是，我们新开发的自组装和种子生长方法适用于低成本大规模制造有序纳米颗粒阵列，并且使用类似方法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。将这种新型的基于纳米颗粒的平台与分子分辨率下对癌细胞力学性质的理解相结合是我们研究的长期目标，这将建立结构-性质-功能关系，从而能够设计用于癌症诊断和预后的新材料和设备，并满足对纳米尺度的新计量学的要求。* 本计划提出了三个具有具体项目的研究领域，即 * 领域1。急性早幼粒细胞白血病（APL）细胞膜的机械检测：*（1a）。用于非粘附APL细胞的微孔的制造;*（1b）.药物处理和未处理的APL细胞系（NB4）细胞的弹性测量;*（1c）。对耐药细胞的治疗性处理和活力评价;*（1d）. APL患者样本的机械检测。*第二区。维甲酸（RA）受体和维甲酸结合相互作用的单分子力谱（SMFS）研究：*（2a）。全反式维甲酸（ATRA）和RA受体之间的结合相互作用;*（2b）。药物-RA受体和配体-RA受体的单个复合物的解结合力。*第三区。用于抗体、膜蛋白和细胞表面标志物检测的纳米结构平台：*（3a）。用于抗表面标志物（CD38）检测的混合纳米颗粒（NP）阵列;*（3b）.局部表面等离子体共振偶联荧光用于定量表面标志物（CD38和CD56）。概述的方案将包括两名研究生和五名本科生的培训。目前的本科生和博士生也将受益于该计划中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的专业知识培训，用于膜表征和传感应用。* * 在生物物理化学和细胞生物学中使用纳米结构和新型材料的全部潜力，预计与先进的显微镜和光谱技术相结合时可以实现。所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以高分辨率成像，并通过单分子力谱法机械探测。作为对传统整体测量的补充，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。

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.