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.
This research further develops leading-edge technologies to obtain physical and physiological measurements from cancer cells. Such physics-based technological innovation, applied methodically in the contexts of basic biological and clinical cancer research, stands to play an important role in furthering our understanding ofthe incredible complexities of cancer. Combined with genetic and environmental information, cell CT and physiological measurements on cancer cells will fill in portions ofthe cancer puzzle whose completion is required for a cure or prevention.
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