Detection of genetic alteration events caused by ionizing radiation in human embryonic stem cells using next-generation sequencing

Wednesday, September 24, 2014 — Poster Session IV
10:00 a.m. –12:00 p.m.	FAES Academic Center	CC	STEMCELL-4

* FARE Award Winner

Authors

• V.T. Nguyen
• I.V. Panyutin
• I.G. Panyutin
• R.D. Neumann

Abstract

Ionizing radiation (IR) is widely employed for medical therapeutic and diagnostic purposes. Although its cytotoxic properties have long been successfully exploited for therapeutics, the biological effects of IR from clinically relevant diagnostic doses remain unclear. We therefore seek to investigate this urgent question as the use of IR-related diagnostic tests continues to rise. A novel model based on human embryonic stem cell (hESC) culture was established and targeted next-generation sequencing of the genomic “hotspot” regions frequently mutated in 50 human cancer genes was performed. Utilizing this deep sequencing approach, we were able to consistently sequence low-, high-dose, and control samples from 4 different hESC cell lines with reliable coverage for data analysis. Single nucleotide variants occurring at very low frequencies (≈2%) were successfully identified using two independent analysis software platforms, confirming the plausibility of our method. We did not detect any differences in the number of genetic alteration events between the low-dose samples and controls, suggesting that in the highly radiosensitive hESCs, diagnostic low-dose IR did not result in a detectable increase in genetic alteration events occurring within the cancer “hotspot” regions. Future extensive studies are warranted to confirm the safety of medical diagnostic procedures involving IR.

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