Adjuvant Vitamin E
and Ovarian Cancer
Alysha Hartman
Mentor: Debbie Mustacich, Ph.D.
Linus Pauling Institute
Oregon State University
Outline
Introduction
• Ovarian Cancer
• Cisplatin
• Vitamin E
Question
Hypothesis
Experimental Design
Results
Summary
Future Studies
F344 Female Rats
Ovarian Cancer
5th leading cause of cancer-related deaths in women
in the U.S.
• Accounts for about 6% of deaths
Highest mortality rate of gynecologic cancers
Most patients have widespread disease at diagnosis
Cisplatin (CDDP)
Platinum containing chemotherapeutic drug
Highly active against a number of cancers:
• Ovarian, Lung, Cervical, Head & Neck, Testicular
Side effects include:
• Nephrotoxicity, ototoxicity, and neurotoxicity
CDDP-Induced Neuropathy (CIPN)
Affects the nerves that carry sensations to the brain
Major dose-limiting adverse side effect of CDDP
Symptoms include:
•
Tingling & burning in hands and feet
•
Tremors
•
Numbness
CDDP-induced Neuropathy & Vitamin E
 Mechanism of CIPN is undetermined.
• Platinum accumulation
 Clinical and histologic features are similar to
those seen in Vitamin E deficiency neuropathy.
(Muller ‘83, Sokol ’88)
 CDDP decreases plasma Vitamin E in humans.
(Weigl ’98)
Vitamin E (RRR-α-Tocopherol)
RRR-α-Tocopherol (α-T): natural form of vitamin
E preferentially retained in the body. (Traber ’05)
Antioxidant role of α-T
Lipid soluble antioxidant
• Found in cell membranes
Protects cellular lipids from oxidation
• lipid peroxidation
Biomarkers of Lipid peroxidation:
• F2-Isoprostanes
• Malondialdehyde (MDA)
Central Question
Can the neurotoxicity of CDDP be mitigated to
allow increased survival without decreased
quality of life for ovarian cancer patients?
Central Hypothesis:
1) CDDP depletes tissue -T by an oxidative stress
mechanism leading to neurotoxicity
2) Adjunct -T will prevent CDDP-mediated -T
depletion, thereby preventing neurologic damage
3) Adjuvant -T will not decrease CDDP anti-tumor
efficacy
Pre-clinical Model of Ovarian
Cancer
Vitamin E (α-T) and Cisplatin (CDDP) Treatment Schedule
Days:
Week 1
Week 4
(Sun, Tue,
Thur)
GROUP
Week 5
(Friday)
Weeks 5-8
(Mon &
Thur)
Weeks 5-8
(Tues &
Fri)
Week 9
TREATMENT
A Saline
IP inject
cells
SC saline
SC saline
SC saline
SC saline
Sacrifice
B α-T
IP inject
cells
SC α-T
SC saline
SC α-T
SC saline
Sacrifice
C Cisplatin
IP inject
cells
SC saline
SC saline
IP CDDP
SC saline
SC saline
IP CDDP
Sacrifice
D αT/Cisplati
n
IP inject
cells
SC α-T
SC saline
IP CDDP
SC α-T
SC saline
IP CDDP
Sacrifice
* Rats treated with CDDP receive an accumulative dose of 18mg / kg
Adjuvant α-T does not alter CDDPinduced weight loss
Adjuvant α-T increases CDDP
Anti-tumor Efficacy
Saline
CDDP
CDDP and α-T
Adjuvant α-T decreases Tumor
Incidence and Multiplicity
Number of Rats Per Group
Tumors
Per Rat
0
<10
10-50 50-100
2/8
6/8
1/12
8/12
3/12
Group A
Group B
100+
Group C
2/16
2/16
1/16
8/16
3/16
Group D
10/16
2/16
2/16
1/16
1/16
Adjuvant α-T prevents tumor- and CDDPinduced depletion of α-T
#
80
αT
αT
P
D
+
P
C
D
αT
in
e
C
D
D
C
αT
P
D
+
D
P
D
C
αT
P
D
D
D
D
P
C
C
αT
in
e
Sa
l
l
on
C
+
P
D
D
C
tr
o
αT
D
P
D
C
αT
l
e
in
Sa
l
+
C
#
40
0
tr
o
0
#
C
D
*#
0
on
40
Sa
l
+
P
D
C
600
400
200
on
tr
ol
α-T (nmol/g)
Sa
lin
e
800
C
αT
P
D
C
D
αT
in
e
C
α-T (nmol/g)
0
#
400
*
Tumors
*#
*
*
*#
20
Adrenal
1400
1200
1000
80
*#
D
C
+
Sa
l
T
ol
D
D
40
D
C
* # *#
0
α-
T
α-
lin
e
Sa
C
on
tr
ol
0
40
*#
*#
ol
40
*
600
400
200
120
on
tr
60
*#
*#
*#
Plasma
80
on
tr
80
600
400
200
α-T (nmol/g)
*#
α-T (nmol/g)
600
400
200
Lung
Kidney
*#
P
D
α-T (mM)
P
α-T (nmol/g)
Liver
* = p<0.05 vs. non-tumor controls; # = p<0.05 vs. saline treated tumor-bearing rats
Adjuvant α-T improves spinal cord
α-T levels compared to CDDP alone
Cervical
40
*#
30
*#
*
10
0
40
30
*
*
*
*
20
10
Lumbar
*
20
10
30
αT
+
D
D
P
D
D
P
C
#
*
#
*
20
10
αT
C
D
D
P
+
D
P
C
D
αT
C
D
D
P
D
D
P
+
αT
C
αT
Sa
lin
e
on
tr
ol
40
0
0
C
Cauda Equina
C
on
tr
ol
Sa
lin
e
30
*
50
α-T (nmol/g)
α-T (nmol/g)
50
40
C
C
D
D
C
P
+
on
tr
ol
Sa
lin
e
αT
D
P
C
D
αT
C
on
tr
ol
Sa
lin
e
0
αT
20
*
Thoracic
50
α-T (nmol/g)
α-T (nmol/g)
50
* = p<0.05 compared to non-tumor controls; # = p<0.05 compared to saline treated tumorbearing rats.
400
*
*
*#
*#
200
αT
P
D
D
P
+
D
D
C
C
αT
Sa
lin
e
0
on
tr
ol
5-Series F2-Isoprostane
(pg/mL)
600
C
+
αT
D
P
D
P
C
D
αT
0
C
D
D
P
+
αT
P
D
D
C
C
D
αT
Sa
lin
e
C
on
tr
ol
0
100
Sa
lin
e
100
*#
*#
ol
*#
*#
*
200
on
tr
200
*
C
*
300
(pg/mL)
*
15S-8-iso-PGF2a
300
(pg/mL)
15R-8-iso-PGF2a
Adjuvant α-T prevents tumor- and
CDDP-induced elevation of plasma
F2-Isoprostanes
* = p<0.05 compared to non-tumor controls; # = p<0.05 compared to saline treated tumorbearing rats.
* = p<0.05 compared CDDP alone
C
DC
DD
PD
+P+
α- α
T -T
P
D
p = 0.07
80.10
6
40.05
2
00.00
-αTT
+α
P+
C CD
D D
D P
P +
+ α
α- -T
T
CC
DD
DD
PP
*
200.15
DC
DD
PD
6
0.05
4
2
0.00
0
0.20
25
C
0.10
8
Tumor
Cauda Equina
DP
D
P
*
0.15
20
P = 0.07
30
Platinum (mg/mg)
25
Platinum (mg/mg)
0.20
0.10
8
60.05
4
20.00
0
C
Lung
Lumbar
30
*
D
DC
PD
C CD
D D
D P
P +
+ α
α- -T
T
0.00
0.15
20
D
CD
0.05
10
25
C
0.10
Thoracic
Liver
300.20
Platinum (mg/mg)
20
Platinum (mg/mg)
*
0.15
0
Platinum (mg/mg)
Platinum
(mg/mg)
Cervical
Kidney
0.20
30
C C
D D
D D
P P
Platinum(mg/mg)
(mg/mg)
Platinum
Adjuvant α-T reduces spinal cord and
lung platinum but not tumor platinum
Summary
-T plus CDDP:
 Increases CDDP anti-tumor efficacy
•
Decreased tumor burden
 Prevents CDDP-induced decreases in tissue and plasma
-T
 Prevents tumor- and CDDP-induced lipid peroxidation
 Decreases accumulation of platinum in spinal cord
tissues but not tumors
Ongoing & Future Studies
Ongoing Studies:
Tissue analysis
• MDA – oxidative stress
• Glutathione – antioxidant
Histology
• proliferation markers
Future Work:
Adjuvant Vitamin C
Clinical Trials
Acknowledgements
Debbie Mustacich, Ph.D.
 Kevin Ahern, Ph.D.
Shannon Pease
 ACS Research Scholar
Michelle Beehler
grant 120548-RSG-11-
Devin Corrigan
075-01-CCE
Giovanna Coto, M.A., DVM
Christiane Löhr, DVM, Ph.D.
F344 Rats
Linus Pauling Institute
URISC
Questions?
Pilot Study in Healthy Rats: CDDP
Decreases Ganglia α-T **
a-T (nmol/g)
50
Veh
Vit E
CDDP
CDDP + VE
b
40
a,b
a
30
b
d
a
c
c
20
10
0
D. Ganglia
V. Ganglia
Ganglia
** Columns within the same tissue but with different letter designations are significantly different, p<
0.05
Lipid Peroxidation
1) Initiation
• Production of a carbon radical
2) Propagation
• Peroxyl radical and an additional
carbon radical are formed
3) Termination
• Two peroxyl radicals combine to
stop the chain reaction
F2-Isoprostane from Arachidonic Acid