Two for One

Novel Technique Reveals Both Gene Number & Protein Expression

Arnab Chakravarti 2Researchers have discovered a fluorescence microscopy technique for simultaneously visualizing gene number and protein expression in individual cells.

The new technique, called the fluorescent in situ gene protein assay, combines traditional fluorescent in situ hybridization (FISH) with the in situ proximity ligation assay, which is capable of resolving individual protein molecules.

The technique could permit a detailed analysis of the relationship between gene status and expression of the corresponding protein in cells and tissues. Study leaders say this may bring a clearer understanding of cancer and other complex diseases.

“To my knowledge, this is the first technique that allows us to concurrently address gene activity and corresponding protein expression in the same cells,” says co-principal investigator Arnab Chakravarti, MD, chair of Radiation Oncology at Ohio State and co-director of the Brain Tumor Program at the OSUCCC – James. “The ability to resolve gene- and protein-expression changes across a tumor could help us understand what drives tumor behavior overall.”

For this study, principal investigator Markus Bredel, MD, PhD, associate professor at the University of Alabama-Birmingham and adjunct associate professor of Radiation Oncology at the OSUCCC – James, along with Chakravarti and their collaborators, first assayed fixed human glioblastoma tumor cells, then paraffin-embedded human glioblastoma tissue. In both cases, they assayed for overexpression of a mutant form of the epidermal growth factor receptor gene, EGFRvIII, and for levels of its truncated protein in glioblastoma.

“This method has potential to perform a detailed analysis of the relationship between cancer gene status and corresponding protein expression in cells and tissues,” Bredel says. “We demonstrate that the fluorescent in situ gene protein assay methodology is capable of resolving cancer gene and protein patterns simultaneously on a cell-by-cell basis, which is particularly important in heterogeneous diseases such as cancer.”

Published in the journal Neuro-Oncology.

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