Dr. Kolja A. Wawrowsky Profile

Dr. Kolja A. Wawrowsky

at Infinicyte Inc

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Proceedings Article | 28 February 2008 Paper

Characterization of tumor cells and stem cells by differential nuclear methylation imaging

Jian Tajbakhsh, Kolja Wawrowsky, Arkadiusz Gertych, Ori Bar-Nur, Eugene Vishnevsky, Erik Lindsley, Daniel Farkas

Proceedings Volume 6859, 68590F (2008) https://doi.org/10.1117/12.769289

KEYWORDS: Tumors, Stem cells, Cancer, Image analysis, 3D image processing, Confocal microscopy, Visualization, Image filtering, Digital filtering, Surgery

Read Abstract +

DNA methylation plays a key role in cellular differentiation. Aberrant global methylation patterns are associated with several cancer types, as a result of changes in long-term activation status of up to 50% of genes, including oncogenes and tumor-suppressor genes, which are regulated by methylation and demethylation of promoter region CpG dinucleotides (CpG islands). Furthermore, DNA methylation also occurs in nonisland CpG sites (> 95% of the genome), present once per 80 dinucleotides on average. Nuclear DNA methylation increases during the course of cellular differentiation while cancer cells usually show a net loss in methylation. Given the large dynamic range in DNA methylation load, the methylation pattern of a cell can provide a valuable distinction as to its status during differentiation versus the disease state. By applying immunofluorescence, confocal microscopy and 3D image analysis we assessed the potential of differential nuclear distribution of methylated DNA to be utilized as a biomarker to characterize cells during development and when diseased. There are two major fields that may immediately benefit from this development: (1) the search for factors that contribute to pluripotency and cell fate in human embryonic stem cell expansion and differentiation, and (2) the characterization of tumor cells with regard to their heterogeneity in molecular composition and behavior. We performed topological analysis of the distribution of methylated CpG-sites (MeC) versus heterochromatin. This innovative approach revealed significant differences in colocalization patterns of MeC and heterochromatin-derived signals between undifferentiated and differentiated human embryonic stem cells, as well as untreated AtT20 mouse pituitary tumor cells compared to a subpopulation of these cells treated with 5-azacytidine for 48 hours.

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