This transnetwork project, awarded by the NCI to a group comprising teams from the Princeton, ASU and Bay Area PS-OCs, seeks to probe differences in chromatin organization between curable and incurable cancers using a variety of assays. (The Princeton component of the project will be executed by Robert Getzenberg, Robert Veltri, Donald Coffey and others at Johns Hopkins University.) As part of this transnetwork collaboration, Project 3 of the ASU PS-OC, led by Dr. Deirdre Meldrum, will quantify the 3D organization of chromatin and select nuclear proteins in fixed cells. We will apply single-cell optical projection computed tomography (Cell-CT) and 3D image analyses to accomplish our goal. Our investigations will be performed on four model cell lines representing curable and incurable cancers provided by the Princeton PSOC. We will establish and maintain cultures of these cell lines in our lab at ASU. We will apply absorption-mode Cell-CT on hematoxylin- or Feulgen-stained cells to probe 3D nuclear structure and chromatin texture. To assess the spatial localization of a select group of nuclear proteins (SATB1, SATB2, Lamins A/C and B, CTCF, HMGA1 and Histone 2A) we will perform fluorescence-mode cell CT imaging on fixed immunolabeled cells. When available, the antibodies and protocols for immunolabeling will be supplied by the Bay Area PSOC. We will debug and validate the immunostaining protocols in our hands using fluorescence confocal microscopy before proceeding to the Cell-CT fluorescence analysis. After Cell-CT fluorescence data acquisition, we will perform 3D image analyses to quantify nuclear structure, chromatin texture, and spatial localization and qualitative expression levels of the nuclear proteins. Our goal is to develop the set of numerical descriptors (3D features and relative expression levels) that best differentiate the 3D nuclear organization, including that of chromatin and of the nuclear proteins, of curable vs. incurable cancers.
|Effective start/end date||8/1/11 → 7/31/12|
- HHS: National Institutes of Health (NIH): $99,904.00