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OMICS Group International is an amalgamation of Open Access publications and worldwide
international science conferences and events. Established in the year 2007 with the sole aim of
making the information on Sciences and technology ‘Open Access’, OMICS Group publishes
500 online open access scholarly journals in all aspects of Science, Engineering, Management
and Technology journals. OMICS Group has been instrumental in taking the knowledge on
Science & technology to the doorsteps of ordinary men and women. Research Scholars,
Students, Libraries, Educational Institutions, Research centers and the industry are main
stakeholders that benefitted greatly from this knowledge dissemination. OMICS International
also organizes 500 International conferences annually across the globe, where knowledge
transfer takes place through debates, round table discussions, poster presentations,
workshops, symposia and exhibitions.
About OMICS Group
About OMICS International Conferences
About OMICS Group
OMICS International is a pioneer and leading science event organizer, which publishes around
500 open access journals and conducts over 300 Medical, Clinical, Engineering, Life Sciences,
Pharma scientific conferences all over the globe annually with the support of more than
1000 scientific associations and 30,000 editorial board members and 3.5 million followers to its
credit.
OMICS International has organized 500 conferences, workshops and national symposiums
across the major cities including San Francisco, Las Vegas, San Antonio, Omaha, Orlando,
Raleigh, Santa Clara, Chicago, Philadelphia, Baltimore, United Kingdom, Valencia, Dubai,
Beijing, Hyderabad, Bengaluru and Mumbai.
4th International Conference and Exhibition on Metabolomics & Systems BiologyApril 29, 2015 – Philadelphia, USA
Inmaculada Martinez-Reyes – Dr. Navdeep Chandel Lab – Northwestern University, Chicago
Dissecting the distinct functions of mitochondria
Mitochondrial metabolism is necessary for tumorigenesis.
Acetyl-CoA
FADH2
FAD
Citrate
α-KG
TCA Cycle
NADH
NAD+
I
II
ATP-Citrate Lyase
III
V
α-KG
H2O2
NFkB HIF ?
Ac
Me
HAT
JMJD3
IV
OAA
Acetyl-CoA
Glucose Pyruvate
H+
H+
H+
H+
O2 H2O
ADP
ATP
Q
Q
e-
e-
Q
e-
c
e-
ce-
e-
Mitochondria as signaling organelles.
CatabolismAMPKAMP/ATP
What functions of mitochondrial metabolism are necessary for
cell proliferation and epigenetics?
H+
ETC
H2OO2
H+
ADP ATP
H+
Two functions of mitochondria: TCA cycle metabolites and membrane potential.
Y(ATP, ROS, Iron-sulfur clusters)
TCA
Metabolites
(Acetyl-CoA, Succinyl-CoA)
NADH/FADH2
NAD+/FAD
DNA polymerase gamma (POLG) is necessary for mitochondrial DNA (mtDNA) replication.
Doxycycline (DOX)
Dominant Negative (DN-POLG)
Dominant Negative (DN-POLG)
Doxycycline
Hans Spelbrink
Doxycycline
Day 0
Day 3
Day 6
Day 9
Day 9
WT PLOG + DOX
DN PLOG + DOX
Dominant negative POLG depletes mtDNA encoded RNA transcripts.
Ty WangJanine Santos
COXII
SDHA
Tubulin
- - - - + + + +
0 3 6 9 0 3 6 9
DOX
Days
WT-POLG
COXII
SDHA
Tubulin
DOX
Days
DN-POLG
- - - - + + + +
0 3 6 9 0 3 6 9
Dominant negative POLG depletes mtDNAencoded proteins.
Loss of mitochondrial DNA does not induce cell death.
D a y 0 D a y 3 D a y 6 D a y 9
0
2 0
4 0
6 0
8 0
1 0 0%
via
bil
ity
W T -P O L G
D N -P O L G
- + + + D O X
Loss of mitochondrial DNA decreases oxygen consumption rate (OCR).
D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5
Re
lati
ve
Mit
oc
ho
nd
ria
l O
CR
to D
ay
0 (
Inta
ct
ce
lls
)
W T -P O L G
D N -P O L G
- + + + D O X
Loss of mitochondrial DNA decreases mitochondrial membrane potential.
D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5M
em
bra
ne
Po
ten
tia
l
(No
rm
ali
ze
d T
MR
E-C
CC
P)
W T -P O L G
D N -P O L G
- + + + D O X
Loss of mitochondrial DNA increasesglucose catabolism.
Biologa -D -G lu c o s e
0
2
4
6
8
1 0
Dy
e R
ed
uc
tio
n
(Arb
itra
ry
Un
its
)
D a y 0
D a y 3
D a y 6
D a y 9
D N -P O L G
Loss of mitochondrial DNA induces dependence on glycolysis for survival.
0
2 0
4 0
6 0
8 0
1 0 0
De
ad
ce
lls
(%
PI+
)
- + - + - + - + G a la c t o s e
+ - + - + - + - G lu c o s e
D N -P O L G
D a y 0 D a y 3 D a y 6 D a y 9
- + + + D O X
Loss of mitochondrial DNA induces AMPK activation.
AMPK
p-AMPK
DN-POLG
Tubulin
DOX
Days
- - + + + + + +
0 0 3 3 6 6 9 9
Loss of mitochondrial DNA diminishes cell proliferation rate.
0
51 0 4
11 0 5
21 0 5
21 0 5
31 0 5
W T -P O L G
D N -P O L G
+ + D O X
Ce
ll n
um
be
r
D a y 3
0
51 0 5
11 0 6
21 0 6
21 0 6
31 0 6
W T -P O L G
D N -P O L G
+ + D O X
Ce
ll n
um
be
r
D a y 6
0
5 .01 0 6
1 .01 0 7
1 .51 0 7
2 .01 0 7
W T -P O L G
D N -P O L G
+ + D O X
Ce
ll n
um
be
r
D a y 9
0
51 0 7
11 0 8
21 0 8
W T -P O L G
D N -P O L G
+ + D O X
Ce
ll n
um
be
r
D a y 1 2
Quantification of Histone modifications using Silac.
Yingming Zhao
He Huang
Minimal changes in methylation and acetylation of H2B and H4.
H3K
9ac
H3K
14ac
H3K
18ac
H3K
27ac
H3K
23ac
H3K
56ac
H3K
79ac
0 .0
0 .5
1 .0
1 .5
Re
lati
ve
ex
pre
ss
ion
(No
rm
ali
ze
d t
o d
ay
0) D a y 0
D a y 3
D a y 6
D N -P O L G
H3K27ac
0 3 6 9
DN-POLG
H3K18ac
DOX- + + +
Days
H3 total
H3K9ac
H3K14ac
Loss of mitochondrial DNA decreases specific histone H3 acetylation.
Mitochondria metabolism is necessary for cell proliferation and specific histone
acetylation.
H+
ETC
H2OO2
H+
ADP ATP
H+
Two functions of mitochondria: TCA cycle metabolites and membrane potential.
Y(ATP, ROS, Iron-sulfur clusters)
TCA
Metabolites
(Acetyl-CoA, Succinyl-CoA)
NADH/FADH2
NAD+/FAD
Electron transport chain couples electron flux to proton pumping.
C-I
C-IIC-III
Q
NADH
NAD+ FADH2
FAD
H+ H+ H+
O2H2O
C-IV
Cyt c
C-I
C-II
C-IIIQ
NAD+ FADH2
FAD
H+ H+ H+
O2H2O
C-IV
Cyt c
NADH
O2H2O
AOXNDI1
NAD+
NADH
NDI1 mimics complex I and AOX bypasses complex III and complex IV.
Eric Dufor
NDI1 and AOX expression uncouples electron flux from proton pumping in cells with depleted mitochondrial DNA.
C-II
Q
NADH
NAD+FADH2
FAD
O2 H2O
NDI1 AOX
NDI/
AOX
H2OO2
ATP4- ADP3-
AOX-NDI1 expression restores electron flux but not membrane potential.
Y
TCA
Metabolites
NADH/FADH2
NAD+/FAD
AOX-NDI expression does not restore mitochondrial DNA.
0 .0
0 .5
1 .0
1 .5
CO
XII
/SD
HA
DN
A r
ati
o
- + + + D O X
D a y 0 D a y 3 D a y 6 D a y 9
D N -P O L G -A O X /N D I1
AOX-NDI expression restores oxygen consumption rate in isolated mitochondria.
Seahorse Biosciences
0 1 0 2 0 3 0
0
2 0 0
4 0 0
6 0 0
T im e (m in )
G F P /B F P D a y 0
A O X /N D I1 D a y 0
P y r + M a l R o t + A n t S H A M
Ox
yg
en
Co
ns
um
pti
on
Ra
te (
pm
ol/
min
)
Pe
rm
ea
bil
ize
d C
ell
s
0 1 0 2 0 3 0
0
1 0 0
2 0 0
3 0 0
T im e (m in )
G F P /B F P D a y 6
A O X /N D I1 D a y 6
P y r + M a l R o t + A n t S H A M
Ox
yg
en
Co
ns
um
pti
on
Ra
te (
pm
ol/
min
)
Pe
rm
ea
bil
ize
d C
ell
s
Study of the metabolic profile of the cells
-2.00 2.00
DN-POLG-GFP/BFP Day 0
DN-POLG-GFP/BFP Day 3
DN-POLG-GFP/BFP Day 6
DN-POLG-GFP/BFP Day 9
DN-POLG-AOX/NDI1 Day 0
DN-POLG-AOX/NDI1 Day 3
DN-POLG-AOX/NDI1 Day 6
DN-POLG-AOX/NDI1 Day 9
Amino Acid
Carbohydrate
Cofactors and Vitamins
Energy
Lipid
Nucleotide
Peptide
Xenobiotics
Super Pathway Sub PathwaySenescent 0d Senescent 0d Senescent 0d Active 0d Active 0d Active 0d
Creatine Metabolism
Oxidative Phosphorylation
Purine and Pyrimidine Metabolism
Pantothenate and CoA MetabolismBiotin MetabolismFolate Metabolism
Pterin Metabolism
Fold of Change
0.75 0.875 1 1.25 1.5 2
Active 6d Active 9dSenescent 3d Senescent 6d Senescent 9d Active 3d
Alanine and Aspartate Metabolism
Glutamate Metabolism
Methionine, Cysteine, SAM and Taurine Metabolism
Urea cycle; Arginine and Proline Metabolism
Aminosugar Metabolism
Long Chain Fatty Acid
Polyunsaturated Fatty Acid (n3 and n6)
Fatty Acid Metabolism
Glycerolipid Metabolism
Energy TCA Cycle
Carbohydrate
Glycolysis, Gluconeogenesis, and Pyruvate Metabolism
Pentose Phosphate Pathw ay
Pentose Metabolism
Nucleotide Sugar
Nucleotide
Purine Metabolism, (Hypo)Xanthine/Inosine containing
Purine Metabolism, Adenine containing
Purine Metabolism, Guanine containing
Pyrimidine Metabolism, Orotate containing
Pyrimidine Metabolism, Uracil containing
Pyrimidine Metabolism, Cytidine containing
Pyrimidine Metabolism, Thymine containing
Cofactors and Vitamins
Nicotinate and Nicotinamide Metabolism
Riboflavin Metabolism
Tetrahydrobiopterin Metabolism
Study of the metabolic profile of the cells
AOX-NDI expression maintains levels of TCA cycle metabolites.
0 .0
0 .5
1 .0
1 .5
Fu
ma
ra
te
Ab
un
da
nc
e (
A.U
)
D N -P O L G D N -P O L G A O X N D I1
D a y 0 D a y 3 D a y 6 D a y 9 D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5
2 .0
2 .5
Cit
ra
te
Ab
un
da
nc
e (
A.U
)
D N -P O L G D N -P O L G A O X N D I1
D a y 0 D a y 3 D a y 6 D a y 9 D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5
Ma
late
Ab
un
da
nc
e (
A.U
)
D N -P O L G D N -P O L G A O X N D I1
D a y 0 D a y 3 D a y 6 D a y 9 D a y 0 D a y 3 D a y 6 D a y 90
1
2
3
4
Alp
ha
-Ke
tog
luta
ra
te
Ab
un
da
nc
e (
A.U
)D N -P O L G D N -P O L G A O X N D I1
D a y 0 D a y 3 D a y 6 D a y 9 D a y 0 D a y 3 D a y 6 D a y 9
AOX-NDI does NOT restore mitochondrial membrane potential.
D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5M
em
bra
ne
Po
ten
tia
l
(No
rm
ali
ze
d T
MR
E-C
CC
P)
D N -P O L G G F P /B F P
D N -P O L G A O X /N D I1
- + + + D O X
NDI/
AOX
H2OO2
ATP4- ADP3-
AOX-NDI1 expression restores electron flux (TCA cycle) but not membrane potential.
Y
TCA
Metabolites
NADH/FADH2
NAD+/FAD
Is electron flux (TCA cycle) sufficient to restore cell proliferation and histone
acetylation?
Electron flux (TCA cycle) is sufficient to restore histone acetylation.
H3K
9ac/H
3
H3K
14ac/H
3
H3K
18ac/H
3
H3K
27ac/H
3
0 .0
0 .5
1 .0
1 .5
2 .0
Re
lati
ve
ex
pre
ss
ion
(No
rm
ali
ze
d t
o d
ay
0) D a y 0
D a y 9
D N -P O L G
H3K
9ac/H
3
H3K
14ac/H
3
H3K
18ac/H
3
H3K
27ac/H
3
0 .0
0 .5
1 .0
1 .5
2 .0
Re
lati
ve
ex
pre
ss
ion
(No
rm
ali
ze
d t
o d
ay
0) D a y 0
D a y 9
D N -P O L G -A O X /N D I10 3 6 9
DN-POLG-AOX/NDI1
DOX- + + +
Days
H3K14ac
H3 total
H3K9ac
H3K27ac
H3K18ac
Electron flux (TCA cycle) is NOT sufficient to restore cell proliferation.
Electron flux (TCA cycle) is NOT sufficient to restore increased glucose catabolism.
a -D -G lu c o s e
0
2
4
6
8
1 0
Dy
e R
ed
uc
tio
n
(Arb
itra
ry
Un
its
)
D a y 0
D a y 6
D N -P O L G -G F P /B F P
a -D -G lu c o s e
0
2
4
6
8
1 0
Dy
e R
ed
uc
tio
n
(Arb
itra
ry
Un
its
)
D a y 0
D a y 6
D N -P O L G -A O X N D I1
Electron flux (TCA cycle) is NOT sufficient to restore glucose dependency for survival.
0
2 0
4 0
6 0
8 0
1 0 0
De
ad
ce
lls
(%
PI+
)
- + - + - + - + G a la c t o s e
+ - + - + - + - G lu c o s e
D N -P O L G -A O X -N D I1
D a y 0 D a y 3 D a y 6 D a y 9
- + + + D O X
DOX
Days
AMPK
p-AMPK
Actin
- - + + + + + +
0 0 3 3 6 6 9 9
DN-POLG-AOX/NDI1
Electron flux without proton pumping is NOT sufficient to alleviate energetic stress.
H+
ETC
H2OO2
H+
ADP ATP
H+
TCA
Two functions of mitochondria: TCA cycle metabolites and membrane potential.
Cell Proliferation
Y?Histone Acetylation
NADH/FADH2
NAD+/FAD
Acknowledgements
Collaborators
He HuangYingming Zhao
Ty WangJanine SantosEric Dufor
Hans SpelbrinkRalph Deberardinis
Chandel Lab
Nav Chandel
Sam WeinbergHeywon Kong
Lauren DieboldJames EisenbartManan MethaColleen Reczek
Arianne RodriguezMichael Schieber
Loss of ATPIF1 allows the maintenance of mitochondrial membrane potential
D a y 0 D a y 3 D a y 6 D a y 9
0 .0
0 .5
1 .0
1 .5
Mit
oc
ho
nd
ria
l M
as
s
(No
rm
ali
ze
d t
o d
ay
0)
W T -P O L G
D N -P O L G
- + + + D O X
Loss of mitochondrial DNA decreased mitochondrial mass.
Loss of mitochondrial DNA alters mitochondrial morphology
0 .0
0 .5
1 .0
1 .5
2 .0
Ac
ety
l-C
oA
Ab
un
da
nc
e (
A.U
)
D N -P O L G D N -P O L G A O X N D I1
D a y 0 D a y 3 D a y 6 D a y 9 D a y 0 D a y 3 D a y 6 D a y 9
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