Project 3

项目3

• 批准号: 8744861
• 负责人: Deirdre R. Meldrum
• 金额: $ 27.09万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8744861
• 关键词: Antibodies; Biological; Biopsy; Cancer Biology; Cancer cell line; Cell Line; Cells; Chromatin; Clinical; Colon; Colon Carcinoma; Computer Simulation; Computer software; Data; Dependence; Elasticity; Esophageal; Feulgen stain; Fluorescence; Functional Imaging; Gel; Gene Expression Profile; Genetic; Genomics; Hematoxylin and Eosin Staining Method; Heterochromatin; Image; Imagery; Instruction; Intervention; Ions; Length; Life; Malignant Neoplasms; Malignant neoplasm of esophagus; Measurement; Measures; Methods; Metric; Modeling; Modification; Nuclear; Nuclear Envelope; Optical Tomography; Optics; Participant; Patients; Physics; Physiological; Play; Ploidies; Prevention; Principal Investigator; Proteins; Proteomics; Protocols documentation; Research; Research Personnel; Research Project Grants; Resolution; Respiration; Role; Sampling; Scanning; Shapes; Staining method; Stains; Statistical Methods; System; Technology; Texture; Thick; Time; Work; absorption; anticancer research; aqueous; base; cancer cell; cancer diagnosis; cell fixation; cell fixing; cell type; density; design; imaging modality; immortalized cell; improved; insight; instrument; instrumentation; novel; physical science; programs; protein expression; seal; spatiotemporal; technological innovation; tomography; tool; transcriptomics; tumor; tumor progression

PROJECT SUMMARY (See instructions): The aim of this research is to further develop and apply the novel tools, 3D optical tomography and labs on chips, to better understand the physical attributes of cancer cells. Deeper understanding of cancer cells' physical attributes will allow their quantitative comparison between cancer grades, and before and after genomic and chemotherapeutic interventions, and is an important component of progress toward cures and preventions. Both of these technologies will be applied to single cells. To present a tractable research plan, the cancer cell types studied are colon and esophageal, but the results may be relevant to other cancers. Attributes of immortalized cell lines representing both cancers and cells from patient biopsies will be characterized. Optical Tomography: 3D optical tomography provides two types of information at the 100-nm length scale: The first, receiving primary emphasis, is morphometric (structural: shape, size, chromatin texture and density), relying on absorption imaging analogous to the type (H&E and Feulgen staining) of imagery that cancer cytopathologists have used for decades to diagnose cancer clinically. The second is functional (localized protein concentration), relying on antibody and other fluorescent staining. Among the novel contributions harnessed for this work is the instrumentation and software to perform cell CT imaging for the first time. This provides truly isotropic resolution on cells suspended in their natural state, eliminating ambiguities due to overlapping features in thick sections and smears, and problems with orientation dependence, distortions due to flattening, and the incomplete sampling inherent in all 2D imaging methods. Isotropic resolution confers two salient advantages on this method for understanding cancer: 1) Measurements are robust and repeatable, as they represent the entire 3D cell, not randomlyselected plane(s). 2) The measurements are exquisitely sensitive to the neoplastic progression status of a cell. This research will produce and derive such parameters as nuclear to cytoplasmic ratio and ploidy with unprecedented precision; and texture features like heterochromatin distribution and granularity scale, and metrics related to the nuclear membrane infoldings, invaginations and protrusions, impossible to measure in 2D, which constitute signatures common to many cancer cells. Labs on Chips: Before and after cancer relevant modifications, physiological parameters including respiration rate, pH, ion fluxes and ATP concentrations, and transcriptome levels will be quantified within and between cells and their sealed microenvironments. Correlations will be explored between physiological and transcriptomic variables, and between these and the morphometric, densitometric, and protein expression level and localization measurements from cell CT. Our measurements, and such correlations or the lack thereof, before and after genetic and chemotherapeutic modifications, will provide new insights into the biological physics of cancer.

项目摘要（请参阅说明）：这项研究的目的是进一步开发和应用新颖的工具，3D光学断层扫描和实验室，以更好地了解癌细胞的物理属性。对癌细胞的物理属性的更深入了解将使它们​​在癌症等级以及基因组和化学治疗干预之前和之后的定量比较，并且是朝着治愈和预防的进步的重要组成部分。这两种技术都将应用于单个细胞。为了介绍可处理的研究计划，研究的癌细胞类型是结肠和食管，但结果可能与其他癌症有关。代表患者活检的癌症和细胞的永生细胞系的属性将被表征。光学断层扫描：3D光学层析成像在100 nm长度尺度上提供两种类型的信息：首先接受主要重点，是形态计量学（结构：形状，大小，染色质纹理和密度），依赖于类型的吸收成像（H＆E和Feulgen染色）（H＆E和Feulgen染色）的成像学家对癌症细胞病理学家的成像进行了诊断为癌症的临床。第二个是功能性的（局部蛋白质浓度），依赖于抗体和其他荧光染色。在这项工作的新颖贡献中，有仪器和软件首次执行细胞CT成像。这为悬浮在自然状态的细胞上提供了真正的各向同性分辨率，从而消除了由于厚部分和涂片的重叠特征以及方向依赖性的问题，由于扁平化而导致的扭曲以及所有2D成像方法中固有的不完整采样。各向同性分辨率赋予了这种理解癌症方法的两个显着优势：1）测量值稳健且可重复，因为它们代表了整个3D细胞，而不是随机选择的平面。 2）测量值对细胞的肿瘤进展状态非常敏感。这项研究将产生并得出诸如核与细胞质比和曲目诸如前所未有的精度之类的参数。以及质地特征，例如异染色质分布和粒度量表，以及与核膜互和相关的指标，不可能在2D中测量，这构成了许多癌细胞常见的特征。芯片上的实验室：癌症相关修饰前后，生理参数，包括呼吸率，pH，离子通量和ATP浓度，以及转录组水平将在细胞内部和之间及其密封的微环境之间进行量化。将在生理和转录组变量之间探索相关性，以及这些相关性以及形态计量学，光密度测定和蛋白质表达水平以及细胞CT的定位测量值。我们的测量以及遗传和化学修饰之前和之后的相关性或缺乏相关性将为癌症的生物物理学提供新的见解。