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DEUTERIUM DEPLETED WATER ALTERS GLUCOSE-DERIVED FATTY
ACID AND CHOLESTEROL SYNTHESIS OF TUMOR CELLS L.G. Boros1,2, A. Kochegarov1, I. Szigeti1, S.T. Lee1, G. Jancso3, Gy. Jákli3, G. Somlyai4
1SIDMAP, LLC; 2UCLA School of Medicine, Los Angeles, CA, USA.; 3Central Research Institute for Physics,
Atomic Energy Research Branch; 4HYD Ltd., Budapest, Hungary
CONTACT: HYD Ltd., H-1215 Budapest, Deák F. u. 51/a, Hungary. LITERATURE: In Vivo. 2000, 14(3): 437-9.; FEBS Lett. 1993, 317(1-2): 1-4. 2nd Annual Conference of the Metabolomics Society, Boston, USA . June 15, 2006
INTRODUCTION
Deuterium (2H) is the heavy stable non-radiating
isotope of hydrogen (1H) that carries one extra
neutron in the atomic nucleus. Therefore
deuterium’s atomic mass is ~ twice of that of 1H.
Hydrogen atoms of water participate in virtually
all ion exchange and substrate product
transport reactions through the cell membrane
and hydrogen also acts as the reducing equivalent
in energy producing as well as reductive
macromolecule synthesis reactions in all living
cells. Deuterium is also involved in epigenetic
events (changes in gene activity that are not
caused by changes in the DNA sequence).
Deuterium depletion of water in cell culture
media or body fluids temporarily decelerates cell
growth in vitro and induces tumor regression in
vivo.
The exact mechanism and the effects of deuterium
depletion on mammalian cell intermediary
metabolism are not fully know.
HYPOTHESES
Deuterium incorporation from common water into DNA
increases its fragility thus accelerates mutations, aging
and cancer.
Deuterium affects the kinetics of reductive synthesis and
the generation of NADP+ thus altering membrane fatty
acid and cholesterol synthesis.
Deuterium alters tricarboxylic acid cycle and
intermediary metabolism by altering carbon flow and the
rate of product synthesis and energy production.
Deuteration of DNA with adjacent nuclear membrane
structures is an important epigenetic event directly
involved in driving oncogenesis to alter gene expression,
replication and growth.
AIM
To determine metabolic flux-modifying effects of
deuterium depleted water (DDW: 100, 50 and 25 ppm) as
compared to normal deuterium-containing water (150
ppm) on [1,2-13C2]-D-glucose metabolism in cultured
pancreatic (MIA-PaCa), lung (H-441) and breast (MCF-7)
ductal carcinoma cells.
METHODS
After 72 hours of incubation with the [1,2-13C2]-D-glucose tracer in
DDW we analyzed its uptake and contributions to lactate
production, glycolysis, RNA ribose, glycogen, cholesterol and long
chain fatty acid synthesis as well as TCA cycle glutamate and 13CO2
release using GC/MS.
13C LABELED GLUCOSE INTERMEDIARY
METABOLISM - MACROMOLECULE
SYNTHESIS
HOHO
HH
OO1212CC
1212CC
1212CC
1212CC
1313CC
1313CC
OHOH
OO
HH
HH
HH
HH
HH
HH
OHOH
OHOH
PP
[[1,2-1,2-1313CC]glucose –6P]glucose –6P
Phosphoglucose isomerase
Phosphofructo kinase
[[1,2-1,2-1313CC]fructose-6-P]fructose-6-P
OHOH
OO1212CC
1212CC
1212CC
1212CC
1313CC
1313CC
OHOH
HH
HH
HH
HH
OHOH
OO
PP
HH
HH
HH
HOHO
OO
OO1212CC
1212CC
1212CC
1212CC
1313CC
1313CC
OHOH
HH
HH
HH
HH
OHOH
OO
PP
HH
HH
HH
HOHO
PP
[[1,2-1,2-1313CC]fructose-1,6-]fructose-1,6-bisPbisP
TriosePhosphateIsomerase
OO1212CC
1212CC
1212CC
OHOHHH
HH
HH
PP
HH
OO
glyceraldehyde-3Pglyceraldehyde-3P
OO
1212CC
1313CC
1313CC
HH
OO
HH
HH
HOHO
PP
[[2,3-2,3-1313CC]dihydroxy]dihydroxy
acetone-Pacetone-P
HH
Aldolase
GlycolysisGlycolysis
m/z330
GlucGluc
Lactate dehydrogenase
1313CC HH33
1313CC
1212COCO--
OO
[[2,3-2,3-1313CC]pyruvate]pyruvate
OO
1313CC HH33
1212COCO--
OO
1313CC OHOHHH
[[2,3-2,3-1313CC]lactate]lactateRELEASED INTO RELEASED INTO
CULTURE MEDIUMCULTURE MEDIUM
OO1313CC
1313CC
1212CC
OHOHHH
HH
HH
PP
HH
OO
[[2,3-2,3-1313CC]glyceraldehyde-3P]glyceraldehyde-3P
OO
1212CC
1313CC
1313CC
HH
OO
HH
HH
HOHO
PP
[[2,3-2,3-1313CC]dihydroxy]dihydroxy
acetone-Pacetone-P
HH
TriosePhosphate Isomerase
GlycolysisGlycolysis
m/z328LactLact
HO
H
O12C
12C
12C
12C
1313CC
1313CC
OH
O
H
H
H
H
H
H
OH
OH
P
[[1,2-1,2-1313CC]glucose –6P]glucose –6P
O12C
12C
12C
12C
1313CC
OH
H
H
H
H
H
OH
OH
P
O
H
[[1-1-1313CC]ribulose-5P]ribulose-5P[[1,2-1,2-1313CC]-6phosphoglucono]-6phosphoglucono
lactonelactone
HO
O-
O12C
12C
12C
12C
1313CC
1313CC
OH
O
H
H
H
H
H
H
OH
OH
P
1313CCO2
LIPID – DNANADP NADPH NADP NADPH
SYNTHESIS
Glucose-6P Dehydrogenase
(G6PDH)
6-phosphogluconate dehydrogenase
Oxidative Pentose CycleOxidative Pentose Cycle
NUCLEIC ACID NUCLEIC ACID
SYNTHESISSYNTHESIS
OO1212CC
1212CC
1212CC
1313CC
OHOH
HH
HH
HH
HH
OHOH
HH
PP
[1-13C]ribose-5P
OO
1212CCHH OHOH
OO1212CC
1212CC
1212CC
1212CC
1313CC
OHOH
HH
HOHO
HH
HH
HH
HH
OHOH
PP
OO
HH
[1-13C]xylulose-5P
Transketolase OO
OO1212CC
1212CC
1212CC
OHOHHH
HH
HH
HHerythrose-3P
1212CC OHOHHH
PP
m/z256RibRib
m/z328LactLact
OHOH
OO1212CC
1212CC
1212CC
1212CC
1212CC
1313CC
OHOH
HH
HH
HH
HH
OHOH
OO
PP
HH
HH
HH
HOHO
[1-13C]fructose-6-P
GLYCOLYSISGLYCOLYSIS
GLUTAMATE
[2,3-13C]pyruvate
Pyruvatedehydrogenase
[5,6-13C]citrate
[4,5-13C]ketoglutarate
Pyruvatecarboxylase
[2,3-13C]ketoglutarate
[2,3-13C]oxaloacetate
Glut
C2-C5
m/z198
CO2CO2
Glut
C2-C5
m/z198
[2,3-13C]pyruvate
Pyruvatedehydrogenase
[1,2-13C]Acetyl-CoA PalmitatePalmitate
m/z270
m/z272 m/z274
CO2
Fatty acid synthase
RESULTS & CONCLUSIONS
Metabolic profiles of tumor cells after 72 hours of DDW treatment
• Deuterium depleted water (DDW) did not significantly alter
glucose uptake, oxidation and glycogen synthesis in any of the cell
lines (Figure 1).
• Pentose cycle flux relative to glycolysis decreased in MIA-PaCa
cells (Figure 2).
• RNA ribose synthesis and turnover also decreased in Mia-PaCa
cells after 25 ppm treatment (Figure 3).
• TCA cycle substrate flux decreased in MCF-7 breast tumor cells
(Figure 4).
• Lignocerate (C:24) and palmitate syntheses were decreased in
MIA-PaCa cells and cholesterol synthesis was decreased in MCF-
7 breast tumor cells (Figure 5).
MIA-PaCa
150 100 50 25 150 100 50 25 150 100 50 25
Deuterium (2H) in water (ppm)
GLUCOSE CONSUMPTION
0
1
2
3
4
5
6
H-441-Lung
MCF-7-Breast
Mil
lig
ram
/72
ho
urs
/mil
lio
n c
ell
s
MIA-PaCa
150 100 50 25 150 100 50 25 150 100 50 25
Deuterium (2H) in water (ppm)
PENTOSE CYCLE FLUX RELATIVE TO GLYCOLYSIS
H-441-Lung
MCF-7-Breast
13C
la
be
led
la
cta
te m
1/m
2 r
ati
o
0
1
2
3
4
5
6
P<0.05
MIA-PaCa
150 100 50 25 150 100 50 25 150 100 50 25
Deuterium (2H) in water (ppm)
RNA ribose synthesis and turnover
H-441-Lung
MCF-7-Breast
13C
la
be
led
rib
ose
fra
cti
on
in
RN
A (
% o
f to
tal)
0
10
20
30
40
50
60
70
80
P<0.05
MIA-PaCa
150 100 50 25 150 100 50 25 150 100 50 25
Deuterium (2H) in water (ppm)
PYRUVATE DEHYDROGENASE FLUX OF THE TCA CYCLE
H-441-Lung
MCF-7-Breast
13C
la
be
led
glu
tam
ate
fra
cti
on
(m
/z1
98
)
0
10
20
30
40
50
60
70
P<0.05
MIA-PaCa
lignocerate C:24
synthesis
150 100 50 25 150 100 50 25 150 100 50 25
Deuterium (2H) in water (ppm)
De novo fatty acid and cholesterol synthesis
MIA-PaCa
palmitate C:16
synthesis
MCF-7-Breast
cholesterol
synthesis
13C
la
be
led
fa
tty a
cid
fra
cti
on
s (
% o
f to
tal)
0
20
40
60
80
100
120
P<0.05
P<0.05P<0.05
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
• Based on this data decreased deuterium to hydrogen ratios regulate sterol and fatty acid precursor synthesis, which likely affects the
rate of divisions and cellular proliferation via the regulation of reductive synthesis and new membrane formation.
• Deuterium depletion in cytoplasmic water may control cancer formation similarly as low deuterium containing mitochondrial matrix
metabolic water use for reductive synthesis, which is the natural intracellular deuterium depleting mechanism to control epigenetic
DNA deuteration as the time requiring event during oncogenesis for mammalian cells.
• Deuterium depletion in water and food may have a well defined role in cancer prevention and to improve public health.
