DNA damage intensity in fibroblasts in a 3-dimensional collagen matrix correlates with the Bragg curve energy distribution of a high LET particle

Abstract

Purpose: The DNA double-strand break (DSB) damage response induced by high energy charged particles on lung fibroblast cells embedded in a 3-dimensional (3-D) collagen tissue equivalents was investigated using antibodies to the DNA damage response proteins gamma-histone 2AX (γ-H2AX) and phosphorylated DNA-PKcs (p-DNA-PKcs).

Materials and methods: 3-D tissue equivalents were irradiated in positions across the linear distribution of the Bragg curve profiles of 307.7 MeV/nucleon, 556.9 MeV/nucleon, or 967.0 MeV/nucleon ⁵⁶Fe ions at a dose of 0.30 Gy.

Results: Patterns of discrete DNA damage streaks across nuclei or saturated nuclear damage were observed, with saturated nuclear damage being more predominant as samples were positioned closer to the physical Bragg peak. Quantification of the DNA damage signal intensities at each distance for each of the examined energies revealed a biological Bragg curve profile with a pattern of DNA damage intensity similar to the physical Bragg curve for the particular energy. Deconvolution microscopy of nuclei with streaked or saturated nuclear damage pattern revealed more details of the damage, with evidence of double-strand breaks radially distributed from the main particle track as well as multiple discrete tracks within saturated damage nuclei.

Conclusions: These 3-D culture systems can be used as a biological substrate to better understand the interaction of heavy charged particles of different energies with tissue and could serve as a basis to model space-radiation-induced cancer initiation and progression.

Keywords:

• heavy ion irradiation
• DNA damage
• DNA double-strand break repair
• 3-D tissue equivalents

Acknowledgements

This research was supported by the Office of Science (BER), U.S. Department of Energy, Grant No. DE-AI02-05ER64048 and NASA, NNJ05HD36G NSCOR. We wish to thank the NSRL physics support team for assistance in the design and execution of the irradiation logistics and Oliver Delgado for transporting and setting up the tissue equivalents for irradiation.

Declaration of interest: The authors do not have any conflict of interests or financial arrangements to disclose.