Comparing DNA enrichment of proliferating cells following administration of different stable isotopes of heavy water

• Published in: Scientific reports Vol. 7; no. 1; pp. 4043 - 12
• Main Authors: Farthing, Don E., Buxbaum, Nataliya P., Lucas, Philip J., Maglakelidze, Natella, Oliver, Brittany, Wang, Jiun, Hu, Kevin, Castro, Ehydel, Bare, Catherine V., Gress, Ronald E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 22.06.2017; Nature Publishing Group; Nature Portfolio
• Subjects: 13/106; 13/31; 631/1647/2196/1379; 631/67/70; 631/92/147; 639/638/11/296; 64/60; 692/308/575; 82/16; 82/56; Animals; Body water; Bone marrow; Cell growth; Cell Line, Tumor; Cell Proliferation; Deoxyadenosine; Deoxyribonucleic acid; Deuterium; Deuterium Oxide - pharmacology; DNA; DNA - chemistry; DNA - genetics; DNA Replication - drug effects; Headspace; Heavy water; Humanities and Social Sciences; Ionization; Isotope Labeling; Isotopes; Kinases; Mass Spectrometry; Mice; multidisciplinary; Quantitation; Radioactive labeling; Science; Science (multidisciplinary); Stable isotopes; Thymus; Water levels
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-04404-2

Deuterated water ( 2 H 2 O) is a label commonly used for safe quantitative measurement of deuterium enrichment into DNA of proliferating cells. More recently, it has been used for labeling proteins and other biomolecules. Our in vitro - in vivo research reports important stable isotopic labeling enrichment differences into the DNA nucleosides and their isotopologues (e.g. deoxyadenosine (dA) M + 1, dA M + 2, dA M + 3), as well as tumor cell proliferation effects for various forms of commercially available stable heavy water ( 2 H 2 O, H 2 18 O, and 2 H 2 18 O). Using an in vitro mouse thymus tumor cell line, we determined that H 2 18 O provides superior DNA labeling enrichment quantitation, as measured by GC-positive chemical ionization (PCI)-MS/MS. In addition, at higher but physiologically relevant doses, both 2 H 2 18 O and 2 H 2 O down modulated mouse thymus tumor cell proliferation, whereas H 2 18 O water had no observable effects on cell proliferation. The in vivo labeling studies, where normal mouse bone marrow cells (i.e. high turnover) were evaluated post labeling, demonstrated DNA enrichments concordant with measurements from the in vitro studies. Our research also reports a headspace-GC-NCI-MS method, which rapidly and quantitatively measures stable heavy water levels in total body water.